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   2014| January-March  | Volume 10 | Issue 37  
    Online since February 7, 2014

 
 
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ORIGINAL ARTICLES
Heartwood extract of Acacia catechu induces apoptosis in human breast carcinoma by altering bax/bcl-2 ratio
Nikhil Baban Ghate, Bibhabasu Hazra, Rhitajit Sarkar, Nripendranath Mandal
January-March 2014, 10(37):27-33
DOI:10.4103/0973-1296.126654  PMID:24695415
Background: The heartwood extract of A. catechu, called pale catechu or "Katha" in Hindi has been widely used in traditional Indian medicinal system. Although various pharmacological properties of this plant had been reported previously, only a few were concerned with the anticancer activity of this plant. Objective: The objective was to assess the in vitro anticancer and apoptosis inducing effect of 70% methanolic extract of "Katha" (ACME) on human breast adenocarcinoma cell line (MCF-7). Materials and Methods: MCF-7 cell line was treated with increasing concentrations of ACME and cell viability was calculated. Flow cytometric methods were used to confirm the apoptosis promoting role of ACME. Morphological changes were then analysed using confocal microscopy. Western blotting was then performed to investigate the expression of apoptogenic proteins and to analyse the activation of caspases. Results: ACME showed significant cytotoxicity to MCF-7 cells with an IC 50 value of 288.85 ± 25.79 μg/ml. Flow cytometric analysis and morphological studies confirmed that ACME is able to induce apoptosis in MCF-7 cells. Furthermore, immunoblot results suggested the pathway of apoptosis induction by increasing Bax/Bcl-2 ratio which results in the activation of caspase-cascade and ultimately leads to the cleavage of Poly adeno ribose polymerase (PARP). Conclusion: These results provide the evidence that ACME is able to inhibit the proliferation of MCF-7 cells by inducing apoptosis through intrinsic pathway.
  10 2,518 22
Curcumin attenuates neurotoxicity induced by fluoride: An in vivoevidence
Chhavi Sharma, Pooja Suhalka, Piyu Sukhwal, Neha Jaiswal, Maheep Bhatnagar
January-March 2014, 10(37):61-65
DOI:10.4103/0973-1296.126663  PMID:24696547
Background: Curcumin (Cur), an active ingredient of turmeric is known to have multiple activities, including an antioxidant property and has been suggested to be useful in treatment of several neurological diseases. Objective: To investigate the neuroprotective effects of Cur to mitigate the effect of the Fluoride (F) induced neurotoxicity in mice brain using the histological and the biochemical parameters. Materials and Methods: Exposure of mice (30 days old male) to F (120 ppm) daily for 30 days. Result and Discussion: Treatment with the F causes an increase in lipid peroxidation (LPO) and also increase in the neurodegenerative cells in the hippocampal sub-regions. Interestingly, co-treatment with Cur (30 mg/kg BW) with F (120 ppm) for 30 days results in significant decreases in LPO with a concomitant decrease in neurodegeneration as compared with those treated with F alone. Conclusion: Our study reveals that Cur is useful in ameliorating degenerative effects of F in mice brain.
  8 2,868 30
Optimization of quercitrin and total flavonoids extraction from Herba Polygoni Capitati by response surface methodology
Fengwei Ma, Yang Zhao, Xiaojian Gong, Yu Xie, Xin Zhou
January 2014, 10(37):57-64
DOI:10.4103/0973-1296.127343  PMID:24914309
Objective: To optimize the conditions for extraction of quercitrin and total flavonoids (TF) from Herba Polygoni Capitati (Touhualiao in Chinese) by using response surface methodology (RSM). Materials and Methods: A central composite design (CCD) was adopted to investigate the effects of three independent variables including solvent composition (%), solvent-material ratio (ml/g) and extraction time (min) on the responses, quercitrin and TF yields. Results: The optimized conditions of the extraction are as follows: Ethanol concentration, 65.63%; solvent-material ratio, 10.55:1 (ml/g); extraction time, 54.33 min. The established mathematical model described the factors of experimental parameters well and provided a statistically accurate prediction of the optimum yields of quercitrin and TF. Conclusion: The experimental values agreed with those predicted by the established mathematical model, thus indicating the suitability of the model employed and the success of RSM in optimizing the extraction conditions.
  8 1,792 24
Role of different types of potassium channels in the relaxation of corpus cavernosum induced by resveratrol
Selvinaz Dalaklioglu, G Ozbey
January-March 2014, 10(37):47-52
DOI:10.4103/0973-1296.126658  PMID:24696545
Background: Resveratrol (RVT), one of the most commonly employed dietary polyphenol, is used in traditional Japanese and Chinese medicine for treatment of cardiovascular diseases. Recently, we have shown that RVT has a potent relaxant effect on rat corpus cavernosum via endothelium-dependent and -independent mechanisms. Objective: The present study addressed the question whether different types of potassium channels are involved in the endothelium-dependent and -independent mechanism of corpus cavernosum relaxation induced by RVT. Materials and Methods: Strips of corpus cavernosum from rats were mounted in an organ-bath system for isometric tension studies. Results: RVT (1-100 μmol/L) produced concentration-dependent relaxation responses in rat corpus cavernosum pre-contracted by phenylephrine. The non-selective potassium channels blocker tetraethylammonium chloride (TEA, 10 mmol/L), ATP-sensitive potassium (K ATP ) channels blocker glibenclamide (10 μmol/L), and inward rectifier potassium (Kir) channels inhibitor barium chloride (BaCl 2 , 30 μmol/L) caused a significant inhibition on the relaxation response to RVT, whereas voltage-dependent potassium channels inhibitor 4-aminopyridine (4-AP, 1 mmol/L), and large conductance calcium-activated potassium (BK Ca ) channels inhibitor iberiotoxin (IbTX, 0.1 μmol/L) did not significantly alter relaxant responses of corpus cavernosum strips to RVT. In addition, relaxant responses to RVT did not significantly inhibited by the combination of selective inhibitors of small and intermediate conductance BK Ca channels (0.1 μmol/L charybdotoxin and 1 μmol/L apamin, respectively). Conclusion: These results demonstrated that endothelial small and intermediate conductance BK Ca channels are not thought to be an important role in RVT-induced endothelium-dependent relaxation of corpus cavernosum. The endothelium-independent corpus cavernosum relaxation induced by RVT is seems to largely depend on Kir channels and K ATP channels in corporal tissue.
  7 1,726 24
Fatty acid composition and preclinical resarches on Anthemis wiedemanniana Fisch. & Mey.: Discovery of a new anti-inflammatory agent
Tuba Mert Gönenç, Esra Küpeli Akkol, Ipek Süntar, Tuğçe Fafal Erdoğan, Bijen Kıvçak
January-March 2014, 10(37):53-60
DOI:10.4103/0973-1296.126660  PMID:24696546
Background: Anthemis species have been used for the treatment of gastrointestinal disorders, hemorrhoid, stomachache and inflammatory diseases in Turkish folk medicine. Anthemis wiedemanniana Fisch. And Mey. is an endemic plant used as painkiller, antispasmodic, sedative and for the treatment of urinary inflammations. Objective: The objective of the present study is to evaluate the anti-inflamatory activity of the extracts of A. wiedemanniana by using in vivo methods. Materials and Methods: Carrageenan-, PGE 2 - and serotonin-induced hind paw edema, 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema and acetic acid-induced increase in capillary permeability models were used for the anti-inflammatory activity assessment. Moreover, the fatty acid composition of A. wiedemanniana was investigated by gas chromatography (GC). Results: n-Hexane, diethyl ether and total sesquiterpene lactone extracts exhibited significant inhibition in carrageenan-induced hind paw edema and acetic acid-induced increase in capillary permeability model. n-Hexane and total sesquiterperne lactone extracts showed anti-inflammatory activity in PGE 2 - and serotonin-induced hind paw edema model. However none of the extracts displayed significant activity in TPA-induced ear edema model in mice. C 4:0 (Buthyric acid), C 20:0 (Arachidic acid) and C 16:1 (Palmitoleik acid) were found to be the major fatty acids in these species. Saturated fatty acids (SFA) were found in higher amounts than monounsaturated fatty acids and polyunsaturated fatty acids. SFAs were determined as 63.17%, UFAs as 20.89% and PuFAs as 15.95%. Conclusion: This study confirms the traditional usage of A. wiedemanniana for inflammatory diseases.
  5 781 8
Effect of different growth stages of Ziziphora clinopodioides Lam. on its chemical composition
Wenhuan Ding, Tao Yang, Feng Liu, Shuge Tian
January 2014, 10(37):1-5
DOI:10.4103/0973-1296.127329  PMID:24914287
Aims: The aim of this study is to monitor the changes in the chemical composition of Ziziphora clinopodioides Lam. throughout nine different growth stages. Materials and Methods: Volatile components such as essential oils were analyzed using the gas chromatography (GC) and GC-mass spectrometry, and the contents of non-volatile components were determined by a visible spectrophotometer. Results: Hydro-distilled essential oil content ranged from a minimum of 1.1% (in the post-flowering stage) to a maximum of 1.8% (in the flowering stage). The essential oils included pulegone, which was the most abundant component (77.48-87.3%), p-menthanone (2.79-12.39%), trans-isopulegone (1.04-2.06%), d-limonene (0.51-3.03%) and eucarvone (1.5-4.48%). The contents of non-volatile components, such as that of total phenolics (TPC), total flavonoids (TFC), total triterpenoids content (TTC) and total free amino acids content (TFAAC) were measured using visible spectrophotometry. In the growing stage, TPC, TFC, TTC and TFAAC were 9.91-12.80 mg/g, 29.84-50.63 mg/g, 0.57-1.41 mg/g and 13.33-28.56 mg/g, respectively. Conclusion: These data can be used as a basis to determine the optimal harvest time of Z. clinopodioide Lam.
  5 1,915 25
Multi-component analysis in sun-dried and sulfur-fumigated Angelicae Sinensis Radix by single marker quantitation and chemometric discrimination
Yajing Lou, Hao Cai, Xiao Liu, Gang Cao, Sicong Tu, Songlin Li, Xiaoqing Ma, Kunming Qin, Baochang Cai
January 2014, 10(37):189-197
DOI:10.4103/0973-1296.127374  PMID:24914303
Background: A new method has been developed for the simultaneous determination of ferulic acid, senkyunolide A, and Z-ligustilide in Angelicae Sinensis Radix before and after sulfur-fumigation using quantitative analysis of multi-components by a single marker (QAMS). Materials and Methods: The feasibility and accuracy of QAMS were checked by the external standard method, and various high-performance liquid chromatographic instruments and chromatographic conditions were investigated to verify its applicability. Using ferulic acid as the internal reference substance, and the contents of senkyunolide A and Z-ligustilide were calculated according to relative correction factors by high-performance liquid chromatography. Meanwhile, the influence of sulfur-fumigation on these chemical components in Angelicae Sinensis Radix were evaluated and discriminated by chromatographic fingerprint and chemometrics. Results: There was no significant difference observed between the QAMS method and the external standard method. Furthermore, sulfur-fumigation reduced the contents of ferulic acid, senkyunolide A, and Z-ligustilide in Angelicae Sinensis Radix by some degree, and the sun-drying and sulfur-fumigation processing could be easily discriminated by chromatographic fingerprint and chemometrics. Conclusion: QAMS is a convenient and accurate approach to analyzing multi-component when reference substances are unavailable, simultaneously, chemometrics is an effective way to discriminate sun-dried and sulfur-fumigated Angelicae Sinensis Radix.
  5 2,005 22
Antimicrobial, antibiofilm and cytotoxic activities of Hakea sericea Schrader extracts
Ângelo Luís, Luiza Breitenfeld, Susana Ferreira, Ana Paula Duarte, Fernanda Domingues
January 2014, 10(37):6-13
DOI:10.4103/0973-1296.127331  PMID:24914310
Background: Hakea sericea Schrader is an invasive shrub in Portuguese forests. Objective: The goal of this work was to evaluate the antimicrobial activity of H. sericea extracts against several strains of microorganisms, including the ability to inhibit the formation of biofilms. Additionally the cytotoxic properties of these extracts, against human cells, were assessed. Materials and Methods: The antimicrobial activity of the methanolic extracts of H. sericea was assessed by disk diffusion assay and Minimum Inhibitory Concentration (MIC) value determination. The antibiofilm activity was determined by quantification of total biofilm biomass with crystal violet. Cytotoxicity was evaluated by hemolysis assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. Results: For Gram-positive bacteria, MIC values of H. sericea methanolic extracts ranged between 0.040 and 0.625 mg/mL, whereas the fruits extract yielded the lowest MIC for several strains of microorganisms, namely, S. aureus, B. cereus, L. monocytogenes and clinical methicillin-resistant S. aureus (MRSA). Stems and fruits extract at 2.5 mg/mL effectively eradicated the biofilm of S. aureus ATCC 25923, SA 01/10 and MRSA 12/10. Regarding leaves extract, hemolysis was not observed, and in the case of stems and fruits, hemolysis was verified only for higher concentrations, suggesting its low toxicity. Fruits extract presented no toxic effect to normal human dermal fibroblasts (NHDF) cells however for concentrations of 0.017 and 0.008 mg/mL this extract was able to decrease human breast adenocarcinoma cells (MCF-7) viability in about 60%, as MTT test results had confirmed. This is a clearly demonstrator of the cytotoxicity of this extract against MCF-7 cells.
  5 1,917 17
Antiasthmatic effects of schizandrae fructus extract in mice with asthma
Hyungwoo Kim, Yong-Tae Ahn, Youn Sook Kim, Su In Cho, Won Gun An
January 2014, 10(37):80-85
DOI:10.4103/0973-1296.127348  PMID:24914313
Background: Schizandrae fructus (SF), the fruit of Schisandra chinensis, has been used for the treatment of cough, wheezing, dry mouth, hepatitis, cardiovascular disease, and as a tonic and astringent in China, Japan, and Korea. Objective: Investigation of the antiasthmatic effects of SF. Materials and Methods: We investigated the effects of SF on airway hyperresponsiveness (AHR) to methacholine, production levels of antigen-specific antibodies, and histopathological changes in the lung tissue in a mouse model (Balb/c) of asthma induced by repeated intranasal instillation of an antigen. Results: SF lowered AHR to methacholine (P < 0.05), antigen-specific immunoglobulin E (IgE) level (P < 0.01), and immune cell infiltration in mice with asthma. Prednisolone (PD) effectively decreased AHR (P < 0.01), total antibody (P < 0.01) and IgE (P < 0.01) levels, and immune cell infiltration. SF and PD did not affect the levels of antigen-specific IgG1 and IgG2a antibodies. Conclusion: Our data suggest that SF has possible application as an antiasthmatic drug. We also suggest that SF could be used as a complementary or alternative medicine to glucocorticoids.
  4 1,892 22
Rapid, sensitive, and validated UPLC/Q-TOF-MS method for quantitative determination of vasicine in Adhatoda vasica and its in vitro culture
Garg Madhukar, Ennus Tajuddin Tamboli, Parveen Rabea, SH Ansari, MZ Abdin, Ahmad Sayeed
January 2014, 10(37):198-205
DOI:10.4103/0973-1296.127375  PMID:24914304
Background: Adhatoda vasica a perennial herb has been used in Ayurvedic and Unani system of medicines since last 2000 years and has been employed for the treatment of respiratory tract ailments. Objective: To develop and validate new, rapid, and highly sensitive high throughput ultra-performance liquid chromatography/quadrupole-time-of-flight mass-spectrometry (UPLC/Q-TOF-MS) method for the quantitative estimation of vasicine in the leaves and to establish in vitro cultures of Adhatoda vasica for production of vasicine. Materials and Methods: The chromatographic separation was achieved on a Waters ACQUITY UPLC TM BEH C8 (100.0 × 2.1 mm; 1.7 μm) column packing using isocratic mobile phase consisting of acetonitrile: 20 mM ammonium acetate (90:10; v/v) in a multiple reactions monitoring mode using the transitions m/z 189.09 → 171.08 for vasicine. Results: The vasicine was eluted at 2.58 ± 0.05 min and established a dynamic range of linearity over the concentration range of 1-1000 ng/ml (r2 = 0.999 ± 0.0005). The lower limit of detection and quantification was 0.68 and 1.0 ng/ml, respectively. There was no significant difference observed in the content of vasicine (0.92-1.04%w/w) among the eleven samples collected from different locations of India. The in vitro cultures developed showed that addition of extra 28 mM KNO 3 and 100 mM NaCl in MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) + benzyladenine (BA) + indole acetic acid (IAA) (1 ppm each) produces faster biomass and higher amount of quinazoline alkaloids. Conclusion: Rapid, efficient, and sensitive UPLC/Q-TOF-MS method was developed for the estimation of vasicine and an efficient protocol for development of in vitro cultures was proposed, which can be used at large scale for industrial production of vasicine using bioreactors.
  4 1,964 22
Source of variation of isoflavone concentrations in perennial clover species
Bronislava Butkute, Butkute Lemežiene, Giedre Dabkeviciene, Valdas Jakštas, Egidijus Vilcinskas, Valdimaras Janulis
January 2014, 10(37):181-188
DOI:10.4103/0973-1296.127373  PMID:24914302
Introduction: Clover has attracted considerable interest not only as a valuable livestock forage plant, but also as an important source of isoflavones. The current study was aimed to assess the variation of concentration of three isoflavones in clover species grown under a cool temperate climate environment in Lithuania. Materials and Methods: Isoflavone contents were quantified in the plant parts of 21 accessions belonging to five perennial species of genus Trifolium (T. pratense, T. repens, T. medium, T. rubens, and T. pannonicum). Daidzein, formononetin, and genistein concentrations were quantified in separate plant parts by reversed-phase high-performance liquid chromatography. The analyte extraction was performed from dried and ground leaves, stems, flowers, and roots. The procedure included acid hydrolysis of isoflavone glycosides to aglycones. Results: According to the averaged sum of the three isoflavones quantified in leaves-stems-flowers, the five clover species ranked as follows: T. medium (7.54-3.62-2.31 mg/g) >T. pratense> T. rubens> T. pannonicum> T. repens (0.191-0.204-0.171 mg/g). The contribution of individual compound to the total isoflavone content depended on the species, accession, and plant part. The major part of the isoflavones is concentrated in leaves or stems; however, there is a great variation also. Conclusion: There exists a large variation in the total as well as in individual concentration of isoflavones among the clover species and plant parts and within species. With regard to isoflavone concentration and variability within species, some accessions of T. medium and T. pratense can be considered a highly promising source of phytoestrogens.
  4 1,480 15
Chemical analysis, antioxidant, antichemotactic and monoamine oxidase inhibition effects of some pteridophytes from Brazil
Juliana MM Andrade, Carolina dos S Passos, Roger R Dresch, Maria Angélica Kieling-Rubio, Paulo Roberto H Moreno, Amélia T Henriques
January 2014, 10(37):100-109
DOI:10.4103/0973-1296.127354  PMID:24914288
Background: Ferns are a group of plants that have been little explored from a chemical and biological perspective but that have interesting potential, occurring in various parts of the world. Objective: This work investigates the chemical profile and the biological effects of ferns from Brazil. Materials and Methods: Analyses were performed using rapid performance liquid chromatography (RP-LC) with a diode array detector (DAD). Extracts were tested for their in vitro antioxidant activity, by the total reactive antioxidant potential method and for their antichemotactic potential, by the Boyden chamber method. Cytotoxic effects were assessed by lactate dehydrogenase levels, while the monoamine oxidase (MAO) assay was carried out using a fluorescence-based method. Results: Different chemical compositions were found for the studied ferns, such as Asplenium gastonis, in which hesperidin was identified in its extract, while A. serra showed the presence of xanthone mangiferin. The most samples with highest antioxidant activity were the Asplenium serra, Lastreopsis amplissima and Cyathea dichromatolepis extracts, at 10 μg/mL. High antichemotactic activity was found for A. serra (94.06%) and Didymochlaena truncatula (93.41%), at 10 μg/mL. The extracts showed no cytotoxicity at the highest concentration. Against MAO-A, D. truncatula (82.61%), Alsophila setosa (82.21%), Cyathea phalerata (74.07%) and C. delgadii (70.32%) were the most active extracts (100 μg/mL). Conclusion: The hypothesis was considered that phenolics and triterpenes are responsible for these pronounced activities.
  4 2,011 16
Stereo and region-selective biosynthesis of two new dihydroartemisinic acid glycosides by suspension-cultured cells of Artemisia annua
Jianhua Zhu, Zihan Zeng, Liyan Song, Yanshan Hu, Wei Wen, Rongming Yu
January 2014, 10(37):110-114
DOI:10.4103/0973-1296.127357  PMID:24914289
Background: The system of plant-cultured cells is one of the optimal systems to investigate biosynthesis pathway and their bioactive intermediates. Objective: To study the biosynthesis of dihydroartemisinic acid (1) by suspension-cultured cells of Artemisia annua. Materials and Methods: Substrate (compound 1) was administered into the suspension-cultured cells of A. annua and co-cultured for 2 days. The methanol extract was separated on various column chromatography methods and the structures of two biosynthesis products were elucidated based on the analysis of 1 H NMR, 13 C NMR, 2D NMR, and ESI-MS. Time-course curve was also established. Furthermore, in vitro antitumor activities of compounds 1-3 against HepG2, K562, and A549 cell lines were evaluated by MTT assay. Results: Two new compounds were obtained, namely 3α-hydroxy-dihydroartemisinic acid-α-D - glucopyranosyl ester (2) and 15-hydroxy-cadin-4-en-12-oic acid-β-d - glucopyranosyl ester (3). The results demonstrated that the cultured cells of A. annua possessed the abilities to stereo-selective hydroxylate and region-selective glycosylate sesquiterpene compounds in a highly efficient manner. Inhibitory effects of compounds 1-3 on proliferation of HepG2, K562, and A549 cell lines in vitro were also investigated. Conclusion: Two new dihydroartemisinic acid glycosides were obtained by stereo- and region-selective biosynthesis with cultured cells of A. annua.
  3 1,261 14
In vitro anti-telomerase activity of novel lycopene-loaded nanospheres in the human leukemia cell line K562
Amir Gharib, Zohreh Faezizadeh
January 2014, 10(37):157-163
DOI:10.4103/0973-1296.127368  PMID:24914298
Background: Lycopene, a plant carotenoid, has potent effects against the various types of cancer cells. To date, the effect of lycopene in the free and encapsulated forms on the telomerase activity in human leukemia cell line K562 have not been investigated. The aim of the present study was to prepare a novel lycopene-loaded nanosphere and compare its anti-telomearse activity in K562 cell line with those of free lycopene. Materials and Methods: The lycopene-loaded nanospheres were prepared by nanoprecipitation method. The lycopene entrapment efficacy was measured by high-performance liquid chromatography (HPLC) method. The anti-proliferation effect of the lycopene in the free and encapsulated forms in the different times (0-72 h) and the different doses (0-100 μg/ml) on K562 cell line was studied using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The changes of telomerase activity, following treatment with the lycopene in the free and encapsulated forms, were detected using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay. Results: The entrapment efficacy of lycopene was 78.5% ± 2. Treatment of the K562 cell line with lycopene, in particular in encapsulated form, resulted in a significant inhibition of the cell growth and increasing of percentage of apoptotic cells. It has also been observed that the telomerase activity in the lycopene-loaded nanospheres-treated cells was significantly inhibited in a dose and time-dependent manner. Conclusion: Our data suggest a novel mechanism in the anti-cancer activity of the lycopene, in particular in encapsulated form, and could be provided a basis for the future development of anti-telomerase therapies.
  3 1,843 25
Silybum marianum oil attenuates oxidative stress and ameliorates mitochondrial dysfunction in mice treated with D-galactose
Shu Yun Zhu, Ying Dong, Jie Tu, Yue Zhou, Xing Hua Zhou, Bin Xu
January 2014, 10(37):92-99
DOI:10.4103/0973-1296.127353  PMID:24914315
Background: Silybum marianum has been used as herbal medicine for the treatment of liver disease, liver cirrhosis, and to prevent liver cancer in Europe and Asia since ancient times. Silybum marianum oil (SMO), a by-product of silymarin production, is rich in essential fatty acids, phospholipids, sterols, and vitamin E. However, it has not been very good development and use. Objective: In the present study, we used olive oil as a control to investigate the antioxidant and anti-aging effect of SMO in D-galactose (D-gal)-induced aging mice. Materials and Methods: D-gal was injected intraperitoneally (500 mg/kg body weight daily) for 7 weeks while SMO was simultaneously administered orally. The triglycerides (TRIG) and cholesterol (CHOL) levels were estimated in the serum. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), monoamine oxidase (MAO), malondialdehyde (MDA), caspase-3, and Bcl-2 were determined in the liver and brain. The activities of Na + -K + -adenosine triphosphatase (ATPase), Ca 2+ -Mg 2+ -ATPase, membrane potential (ΔΨm), and membrane fluidity of the liver mitochondrial were estimated. Results: SMO decreased levels of TRIG and CHOL in aging mice. SMO administration elevated the activities of SOD, GSH-Px, and T-AOC, which are suppressed by aging. The levels of MAO and MDA in the liver and brain were reduced by SMO administration in aging mice. Enzyme linked immunosorbent assay showed that SMO significantly decreased the concentration of caspase-3 and improved the activity of Bcl-2 in the liver and brain of aging mice. Furthermore, SMO significantly attenuated the D-gal induced liver mitochondrial dysfunction by improving the activities of Na + -K + -ATPase, Ca 2+ -Mg 2+ -ATPase, membrane potential (ΔΨm), and membrane fluidity. Conclusion: These results indicate that SMO effectively attenuated oxidative damage and improved apoptosis related factors as well as liver mitochondrial dysfunction in aging mice.
  3 2,257 21
In vitro and in silico studies on the anticancer and apoptosis-inducing activities of the sterols identified from the soft coral, subergorgia reticulata
Kuniyil Byju, Vattoni Anuradha, Gopalakrishnapai Vasundhara, S Muraleedharan Nair, N Chandramohana Kumar
January 2014, 10(37):65-71
DOI:10.4103/0973-1296.127345  PMID:24914311
Background: Gorgonians and other octocorals are known to possess a huge array of secondary metabolites in which sterols are the major group of secondary metabolites apart from sesquiterpenes and diterpenes, and the bioactive metabolites could show marked biomedical potential for future drug discovery. Objective : This study was intended for the isolation and identification of sterols from the octocoral Subergorgia reticulata and to evaluate the anticancer and apoptosis-inducing activities of the identified sterols through in vitro and in silico approach. Materials and Methods : The organism was collected from Lakshadweep Island. The isolated sterols were identified using Gas chromatography-mass spectrometry (GC-MS). The structure was confirmed by using comparison of their spectra those in National Institute of Standard Technology (NIST) library. The apoptosis inducing effect of identified sterols were determined by PASS online prediction. In vitro cytotoxity studies were carried out using Dalton's lymphoma ascites cells (DLA) and the cell viability was determined by trypan blue exclusion method. Results : Six sterols were identified from the soft coral S. reticulata. They are Cholesta-5,22-diene-3ol (3β), Ergosta-5-22-dien-3ol (3β,22E 24S), Cholesterol, 26,26-Dimethyl-5,24(28)-ergostadien-3β-ol. β-sitosterol, and Fucosterol. In silico predictions showed that the identified sterols exhibited remarkable apoptosis agonist activity. The probability of apoptosis agonist activity were found maximum for 26,26-Dimethyl-5,24 (28)-S. reticulata sterol fractions isolated were found to be having anticancer activity. Conclusions : These findings suggest that S. reticulata contained biologically active sterol compounds that may be useful in the treatment of cancer.
  3 1,884 21
Simultaneous quantification of nine major active components in traditional Chinese prescription Mahuang decoction and the influence of herbal compatibility on their contents
Yu He, Ying Zhu, Ruping Zhang, Lijun Ge, Haitong Wan
January 2014, 10(37):72-79
DOI:10.4103/0973-1296.127346  PMID:24914312
Background: Mahuang decoction (MHD), a famous classic traditional Chinese formula, has been extensively applied for treating cold, influenza, asthma, acute bronchitis, and other pulmonary diseases. However, the interaction among four drugs of MHD has not been clearly deciphered from the aspect of molecular composition. Objective: To assess the quality of MHD and explore the interplay among different prescription drugs. Materials and Methods: A reversed-phase high performance liquid chromatography (RP-HPLC) coupled with diode array detector (DAD) method for the simultaneous separation and determination of nine bioactive components was developed. A somatomedin A (SMA)-phenyl column (4.6 mm Χ 250 mm, 5 μm) was eluted by a gradient mobile phase contained acetonitrile and 0.05% formic acid-0.05% triethylamine aqueous solution. Four detection wavelengths (210, 252, 278, and 291 nm) were utilized for the quantitative analysis due to the different ultraviolet (UV) spectra of these compounds. Results: Satisfactory separation was obtained for all the components, and the assay was fully validated in respects of linearity, precision, stability, and accuracy. It was found that the calibration curves for all analytes showed good linearity (R 2≥ 0.9991) within the test ranges. The relative standard deviations (RSDs) for intra- and interday repeatability were not more than 1.70 and 2.66%, respectively. The spike recoveries of nine components varied from 97.50 ± 1.69 to 99.27 ± 1.37%. Conclusion: The established method was successfully applied to analyze nine active compounds in decoction samples of various drug compatibilities of MHD. The variations of contents were obvious for different combinations, which hinted the mutual promotion or inhibition of componential dissolution among four herbs of MHD.
  2 2,010 22
Protective effects of Aegle marmelos fruit pulp on 2,4,6-trinitrobenzene sulfonic acid-induced experimental colitis
Rohit R Ghatule, Manish K Gautam, Shalini Goel, Amit Singh, Vinod K Joshi, Raj K Goel
January 2014, 10(37):147-152
DOI:10.4103/0973-1296.127366  PMID:24914296
Background: Aegle marmelos (AM) fruit has been advocated in indigenous system of medicine for the treatment of various gastrointestinal disorders, fever, asthma, inflammations, febrile delirium, acute bronchitis, snakebite, epilepsy, leprosy, myalgia, smallpox, leucoderma, mental illnesses, sores, swelling, thirst, thyroid disorders, tumours and upper respiratory tract infections. Objective: The objective of this study was to study the curative effect of 50% ethanol extract of dried fruit pulp of AM (AME) against 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis. Materials and Methods: AME (200 mg/kg) was administered orally, once daily for 14 days after TNBS-induced colitis. Rats were given intracolonic normal saline or TNBS alone or TNBS plus oral AME. AME was studied for its in vitro antibacterial activity against Gram-negative intestinal bacteria and on TNBS-induced changes in colonic damage, weight and adhesions (macroscopic and microscopic), diarrhea, body weight and colonic levels of free radicals (nitric oxide and lipid peroxidation), antioxidants (superoxide dismutase, catalase and reduced glutathione) and pro-inflammatory marker (myeloperoxidase [MPO]) in rats. Results: AME showed antibacterial activity against intestinal pathogens and decreased colonic mucosal damage and inflammation, diarrhea, colonic free radicals and MPO and enhanced body weight and colonic antioxidants level affected by TNBS. The effects of AME on the above parameters were comparable with sulfasalazine, a known colitis protective drug (100 mg/kg, oral). Conclusion: AME shows curative effects against TNBS-induced colitis by its antibacterial activity and promoting colonic antioxidants and reducing free radicals and MPO-induced colonic damage.
  2 1,765 20
Acridone alkaloids with cytotoxic and antimalarial activities from Zanthoxylum simullans Hance
Chao Wang, Jinfu Wan, Zhinan Mei, Xinzhou Yang
January-March 2014, 10(37):73-76
DOI:10.4103/0973-1296.126669  PMID:24696549
Background: Zanthoxylum simullans Hance is a popular natural spice belonging to the Rutaceae family and it is one of the common prescribed herbs in traditional Chinese medicine. Materials and Methods: The chemical constituents were mainly isolated and purified by silica gel column chromatography and semi-preparative High Performance Liquid Chromatography. Their structures were identified by comparing the spectral data with those reported in the literature. Cytotoxic activities for the isolated acridone alkaloids were evaluated against two prostate cancer cell lines PC-3M and Lymph Node Carcinoma of Prostate (LNCaP), and their antimalarial activities were tested against two different strains of the parasite Plasmodium falciparum 3D7, and Dd2. Results: The root bark MeOH extract of Z. simullans Hance afforded β-sitosterol, 4-methoxy benzoic acid, daucosterol, and five acridone alkaloids, normelicopidine, normelicopine, melicopine, melicopidine, and melicopicine. All five acridone alkaloids were isolated from this plant for the first time and exhibited certain cytotoxic and antimalarial activities in vitro. Conclusion: Normelicopidine was the most active against PC-3M, LNCaP and Dd2 with IC 50 values of 12.5, 21.1, and 18.9 ug/mL respectively.
  2 1,398 19
Optimal inductive and cultural conditions of Polygonum multiflorum transgenic hairy roots mediated with Agrobacterium rhizogenes R1601 and an analysis of their anthraquinone constituents
Bing Huang, Huanjie Lin, Chuanyan Yan, Hongyan Qiu, Lipeng Qiu, Rongmin Yu
January-March 2014, 10(37):77-82
DOI:10.4103/0973-1296.126671  PMID:24696550
Background: Polygonum multiflorum is an important medicinal plant. Hairy roots systems obtained by transforming plant tissues with the natural genetic engineer Agrobacterium rhizogenes can produce valuable biological active substances, which have immense potential in the pharmaceutical industry. Objective: To optimize the inductive and cultural conditions of P. multiflorum hairy roots and to identify the major active secondary metabolites in hairy roots. Materials and Methods: P. multiflorum hairy root were mediated with A. rhizogenes R1601 to induce hairy roots. Four combinations, including Murashige-Skoog (MS), 1/2 MS, B 5 , and White, were investigated to optimize the culture medium. MS medium was selected for the growth measurement. The qualitative and quantitative determinations of free anthraquinone in hairy roots were compared with the calli and aseptic plantlets using high-performance liquid chromatography. Results: The inductive rates of hairy roots by leaves were higher than for any other explants. The presence of agropine in the P. multiflorum hairy roots confirmed that they were indeed transgenic. MS medium was the most suitable of the four media for hairy root growth. Meanwhile, the growth kinetics and nutrient consumption results showed that the hairy roots displayed a sigmoidal growth curve and that their optimal inoculation time was 18-21 days. The determination of the anthraquinone constituents indicated that the rhein content of the hairy roots reached 2.495 μg g−1 and was 2.55-fold higher than that of natural plants. Conclusion: Transgenic hairy roots of P. multiflorum could be one of the most potent materials for industrial-scale production of bioactive anthraquinone constituents.
  2 1,622 26
Antidepressant-like activity of Chaihu-Shugan-San aqueous extract in rats and its possible mechanism
Sue Wang, Suiyu Hu, Chunhu Zhang, Juan Qiu, Yunhui Li
January 2014, 10(37):50-56
DOI:10.4103/0973-1296.127342  PMID:24914308
Background: Chaihu-Shugan-San (CHSGS), a traditional Chinese medicinal formula, is commonly used for the treatment of depression in China. However, the molecular mechanism underlying its antidepressant action is unknown. Objective: The objective of this study is to evaluate the antidepressant-like effects of CHSGS and further explore the possible molecular mechanism implicated in its actions. Materials and Methods: The rats were randomly divided into four groups: The normal control group, the model control group, the CHSGS group and the fluoxetine control group. The antidepressant-like effects of CHSGS aqueous extract were assessed in rats exposed to chronic mild stress (CMS) using the open-field test and sucrose water consumption test, its underlying mechanism of anti-depression was explored by determining the effect of CHSGS on the extracellular signal-regulated kinase (ERK) and phospho-ERK (P-ERK) in the hippocampus using western blot. The aqueous extract of CHSGS at a dose of standard (5.9 g/kg·d) was administered intragastrically for 14 days during the CMS model while the fluoxetine control group was given at the same time using fluoxetine hydrochloride (1.8 mg/kg·d). Results: The stressed rats demonstrated decreased locomotor activity in open field test and reduction in sucrose consumption and decreased levels of P-ERK1/2 and the ratio of P-ERK1/2 to total ERK1/2 in the hippocampus. CHSGS alleviated the depressive-like behaviors and increased levels of P-ERK1/2 and the ratio of P-ERK1/2 to total ERK1/2 in stressed rats as well as fluoxetine. Conclusion: In summary, these results suggest that CHSGS aqueous extract possesses an antidepressant-like activity in CMS induced depression model rats, which might be mediated, at least in part, by reversing the stress-induced disruption of ERK activity.
  2 2,046 24
Element analysis and characteristic identification of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. using microwave digestion-inductively coupled plasma atomic emission spectrometry combined with Fourier transform infrared spectrometry
Yajing Lou, Hao Cai, Xiao Liu, Sicong Tu, Ke Pei, Yingying Zhao, Gang Cao, Songlin Li, Kunming Qin, Baochang Cai
January 2014, 10(37):30-36
DOI:10.4103/0973-1296.127337  PMID:24914306
Background: Sulfur-fumigation may induce chemical transformation of traditional Chinese medicines leading to harmful effects following patient ingestion. For quality control, it is urgently needed to develop a reliable and efficient method for sulfur-fumigation identification. Materials and Methods: The spectrochemical identification of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. was carried out to evaluate inorganic elements and organic components. The concentrations of 12 elements, including Zn, Mn, Cu, Fe, Li, Mg, Sr, Pb, As, Cd, Hg, and S of samples were determined by microwave digestion - inductively coupled plasma atomic emission spectrometry (ICP-AES). Meanwhile, Fourier transform infrared spectrometry (FTIR) was used for the study of chemical group characteristic reactions after sulfur-fumigation. Results: The concentrations of Fe, Mg, Hg, and S elements showed significant differences between non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. The characteristic stretching vibrations of some groups in FTIR spectra, such as -OH, -S = O and -S-O, provided the identification basis for the discrimination of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. Conclusion: The application of microwave digestion - ICP-AES was successfully used in combination with FTIR to authenticate and evaluate the quality of medicinal Fritillaria thunbergii Miq. Further applications of this technique should be explored.
  2 1,889 16
Effect of various absorption enhancers based on tight junctions on the intestinal absorption of forsythoside A in Shuang-Huang-Lian, application to its antivirus activity
Wei Zhou, Xuan Xuan Zhu, Ai Ling Yin, Bao Chang Cai, Hai Dan Wang, Liuqing Di, Jin Jun Shan
January-March 2014, 10(37):9-17
DOI:10.4103/0973-1296.126651  PMID:24695554
Background: Forsythoside A (FTA), one of the main active ingredients in Shuang-Huang-Lian (SHL), possesses strong antibacterial, antioxidant and antiviral effects, and its pharmacological effects was higher than that of other ingredients, but the absolute bioavailability orally was approximately 0.72%, which was significantly low, influencing clinical efficacies of its oral preparations seriously. Materials and Methods: In vitro Caco-2 cell and in vivo pharmacokinetics study were simultaneously performed to investigate the effects of absorption enhancers based on tight junctions: sodium caprate and water-soluble chitosan on the intestinal absorption of FTA, and the eventual mucosal epithelial damage resulted from absorption enhancers was evaluated by MTT test and morphology observation, respectively. The pharmacological effects such as antivirus activity improvement by absorption enhancers were verified by MDCK damage inhibition rate after influenza virus propagation. Results: The observations from in vitro Caco-2 cell showed that the absorption of FTA in SHL could be improved by absorption enhancers. Meanwhile, the absorption enhancing effect of water-soluble chitosan may be almost saturable up to 0.0032% (w/v), and sodium caprate at concentrations up to 0.64 mg/mL was safe, but water-soluble chitosan at different concentrations was all safe for these cells. In pharmacokinetics study, water-soluble chitosan at dosage of 50 mg/kg improved the bioavailability of FTA in SHL to the greatest extent, and was safe for gastrointestine from morphological observation. Besides, treatment with SHL with water-soluble chitosan at dosage of 50 mg/kg prevented MDCK damage after influenza virus propagation better significantly than that of control. Conclusion: Water-soluble chitosan at dosage of 50 mg/kg might be safe and effective absorption enhancer for improving the bioavailability of FTA and the antivirus activity in vitro in SHL.
  2 1,929 28
Optimization of genetic transformation of Artemisia annua L. Using Agrobacterium for Artemisinin production
Elfahmi , Sony Suhandono, Agus Chahyadi
January 2014, 10(37):176-180
DOI:10.4103/0973-1296.127372  PMID:24914301
Background: Artemisinin, a sesquiterpene lactone endoperoxide isolated from the medicinal plant Artemisia annua L., is a choice and effective drug for malaria treatment. Due to the low yield of artemisinin in plants, there is a need to enhance the production of artemisinin from A. annua and biotechnological technique may be one of the methods that can be used for the purpose. Aim: To study the transformation efficiency of Agrobacterium tumefaciens in A. annua that could be applied to enhance the production of artemisinin by means of transgenic plants. Setting and Designs: The factors influencing Agrobacterium-mediated transformation of A. annua were explored to optimize the transformation system, which included A. tumefaciens strain and effect of organosilicone surfactants. Three strains of A. tumefaciens, that is, LBA4404, GV1301, and AGL1 harboring the binary vector pCAMBIA 1303 have been used for transformation. The evaluation was based on transient β-glucuronidase (GUS). Materials and Methods: Plant cell cultures were inniatiated from the seeds of A. annua using the germination Murashige and Skoog medium. A. tumefaciens harboring pCAMBIA were tranformed into the leaves of A.annua cultures from 2-week-old-seedling and 2-month-old-seedling for 15 min by vacuum infiltration. Transformation efficiency was determinated by measuring of blue area (GUS expression) on the whole leaves explant using ImageJ 1.43 software. Two organosilicon surfactants, that is, Silwet L-77 and Silwet S-408 were used to improve the transformation efficiency. Results: The transformation frequency with AGL1 strain was higher than GV3101 and LBA4404 which were 70.91, 49.25, and 45.45%, respectively. Effect of organosilicone surfactants, that is, Silwet L-77 and Silwet S-408 were tested on A. tumefaciens AGL1 and GV3101 for their level of transient expression, and on A. rhizogenes R1000 for its hairy root induction frequency. For AGL1, Silwet S-408 produced higher level of expression than Silwet L-77, were 2.3- and 1.3-fold, respectively. For GV3101, Silwet L-77 was still higher than Silwet S-408, were 1.5- and 1.4-fold, respectively. However, GV3101 produced higher levels of expression than AGL1. The area of GUS expression spots of AGL1, LBA4404, and GV3101 strains was 53.43%, 41.06%, and 30.51%, respectively. Conclusion: A. tumefaciens AGl1 strain was the most effective to be transformed in to A. annua than GV3101 and LBA4404 strain. Surfactant Silwet S-408 produced the highest efficiency of transformation.
  1 2,633 25
Gas chromatography-mass spectroscopy analysis of bioactive petalostigma extracts: Toxicity, antibacterial and antiviral activities
FR Kalt, IE Cock
January 2014, 10(37):37-49
DOI:10.4103/0973-1296.127338  PMID:24914307
Background: Petalostigma pubescens and Petalostigma triloculare were common components of pharmacopeia's of multiple Australian Aboriginal tribal groupings which traditionally inhabited the areas in which they grow. Among these groups, they had a myriad of medicinal uses in treating a wide variety of bacterial, fungal and viral infections. This study was undertaken to test P. pubescens and P. triloculare leaf and fruit extracts for the ability to inhibit bacterial and viral growth and thus validate Australian Aboriginal usage of these plants in treating bacterial and fungal diseases. Materials and Methods: P. pubescens, and P. triloculare leaves and fruit were extracted and tested for antimicrobial, antiviral activity and toxicity. The bioactive extracts were further examined by RP-HPLC and GC-MS to identify the component compounds. Results: The methanol, water and ethyl acetate leaf and fruit extracts of displayed potent antibacterial activity. The methanol and ethyl acetate extracts displayed the broadest specificity, inhibiting the growth of 10 of the 14 bacteria tested (71%) for the leaf extract and 9 of the 14 bacteria tested (64%) for the fruit extracts. The water extracts also had broad spectrum antibacterial activity, inhibiting the growth of 8 (57%) and 7 (50%) of the 14 bacteria tested, respectively. All antibacterial extracts were approximately equally effective against Gram-positive and Gram-negative bacteria, inhibiting the growth of 50-75% of the bacteria tested. The methanol, water and ethyl acetate extracts also displayed antiviral activity in the MS2 plaque reduction assay. The methanol and water extracts inhibited 26.6-49.0% and 85.4-97.2% of MS2 plaque formation, respectively, with the fruit extracts being more potent inhibitors. All ethyl acetate extracts inhibited 100% of MS2 plaque formation. All extracts were also non-toxic or of low toxicity. Analysis of these extracts by RP-HPLC showed that the P. triloculare ethyl acetate fruit extract was the least complex of the bioactive extracts. Subsequent analysis of this extract by GC-MS revealed that it contained 9 main compounds: acetic acid; 2,2-dimethoxybutane; 4-methyl-1,3-dioxane; decane; unadecane; 2-furanmethanol; 1,2-benzenediol; 1,2,3-benzenetriol; and benzoic acid. Conclusion: These studies validate Australian Aboriginal therapeutic usage of Petalostigma species and indicate their medicinal potential.
  1 2,010 22
A validated high performance liquid chromatograph-photodiode array method for simultaneous determination of 10 bioactive components in compound hongdoushan capsule
Liancai Zhu, Xian Yang, Jun Tan, Bochu Wang, Xue Zhang
January-March 2014, 10(37):83-88
DOI:10.4103/0973-1296.126673  PMID:24696551
Background: The compound Hongdoushan capsule (CHC) is widely known as compound herbal preparation and is often used to treat ovarian cancer and breast cancer, and to enhance the body immunity, etc., in clinical practice. Objective: To determine simultaneously 10 bioactive components from CHC, namely glycyrrhetinic acid, liquiritin, glycyrrhizin, baccatin III, 10-deacetylbaccatin III, cephalomannine, taxol, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rb1. Materials and Methods: A high performance liquid chromatograph method coupled with photodiode array detector was developed and validated for the 1 st time. Chromatographic analysis was performed on a SHIMADZU C 18 by utilizing a gradient elution program. The mobile phase was acetonitrile (A)-water (B) at a flow rate of 0.8 mL/min. Results: The calibration curve was linear over the investigated concentration ranges with the values of r 2 higher than 0.9993 for all the 10 bioactive components. The average recovery rates range from 98.4% to 100.5% with relative standard deviations ≤2.9%. The developed method was successfully applied to analyze 10 compounds in six CHC samples from different batches. In addition, the herbal sources of 32 chromatographic peaks were identified through comparative studying on chromatograms of standard, the respective extracts of Hongdoushan, RenShen, GanCao, and CHC. Conclusion: All the results imply that the accurate and reproducible method developed has high separation rate and enables the determination of 10 bioactive components in a single run for the quality control of CHC.
  1 1,376 19
Aliphatic and eudesmalolide esters extracted from the roots of Inula racemosa Hook
Maria Khan, SR Mir, Abuzar Ali, Mohammad Yusuf, Mohammed Ali
January-March 2014, 10(37):40-46
DOI:10.4103/0973-1296.126657  PMID:24695458
Background: Phytochemical investigation of hydroalcoholic extract of the root of Inula racemosa Hook. Materials and Methods: Open silica gel column chromatographic techniques with different solvent systems was used for isolation of aliphatic and eudesmalolide esters from hydroalcoholic extract of the root of Inula racemosa. The structure elucidation of the compounds was done on the basis of spectral data analysis, chemical reactions and comparision with literature data. Results: Phytochemical investigation of the hydroalcoholic extract of the root of Inula racemosa Hook. f. led to the isolation of (5z, 13z)-n-decanyl-n-docos-5, 13-dienoate, a new fatty acid ester, two new sesquiterpenic ester identified as 15-[(13z, 18'z, 20'z)-n-tricos-13, 18, 20-trienyl]-eudesmal-4 (11), 6, 12 (13)-trien-8,14-olide-15-oate and 15- [(16'z), (21'z)-n- tetracos-16', 21'- dienyl]-eudesmal-4 (11) 6, 12 (13)-trien-8, 14-olide-15-oate, two new eudesmanolide ester i.e. 15-[(16z)-n-monadec-16'- enyl]-eudesmal-4 (11) 6, 12 (13)-trien-8,14-olide-15-oate and 15-[(16'z)-n-tetracos-16'- enyl]-endesmal-4 (11), 6, 12 (13)-trien-8,14-olide-15-oate along with the known compound n-Hexadecanyl n-docosanoate. Conclusion: Five new phytoconstituents were identified along with one known compound as aliphatic and eudesmalolide esters from the hydroalcoholic extract of the root of Inula racemosa, as mentioned above.
  1 1,318 15
Cytotoxic activity of Alpinia murdochii Ridl.: A mountain ginger species from Peninsular Malaysia
Kae Shin Sim, Halijah Ibrahim, Sri Nurestri Abdul Malek, Devi Rosmy Syamsir, Khalijah Awang
January-March 2014, 10(37):70-72
DOI:10.4103/0973-1296.126666  PMID:24695515
Background: Alpinia murdochii (Zingiberaceae) is a wild ginger species restricted to mountain areas of Peninsular Malaysia. Due to rapid development and deforestation activities, this species is becoming rare. This is the first report of the cytotoxic activity of A. murdochii. Objective: The present study aimed to investigate the cytotoxic effect of leaves and rhizomes of A. murdochii against selected human cancer cell lines by using in vitro cytotoxicity assay. Materials and Methods: The leaves and rhizomes of A. murdochii were extracted in hexane, dichloromethane (CH 2 Cl 2 ), and methanol (MeOH) prior to cytotoxic activity assessment against selected human cancer cell lines, namely MCF7 (hormone dependent breast carcinoma cell line), HT29 (colon carcinoma cell line), and SKOV-3 (ovarian cancer cell line) by using in vitro neutral red cytotoxicity assay. Results: The hexane and CH 2 Cl 2 extracts of both leaves and rhizomes exhibited remarkable cytotoxic effect against SKOV-3 cells with the IC 50 values in the range of 5.2-16.7 μg/ml. Conclusion: Based on the preliminary data obtained in the present study, the leaves and rhizomes of A. murdochii may be viable therapeutic or preventive candidates for the treatment of ovarian cancer.
  1 1,451 17
Dichlorodiaportinol A - A new chlorine-containing isocoumarin from an endophytic fungus Trichoderma sp. 09 from Myoporum bontioides A. Gray and its cytotoxic activity
Chunyuan Li, Bing Gong, Daniel G Cox, Caili Li, Jinhua Wang, Weijia Ding
January 2014, 10(37):153-158
DOI:10.4103/0973-1296.127367  PMID:24914297
Background: Myoporum bontioides A. Gray, an evergreen shrub from the Myoporaceae family, is a commonly used medicinal plant. Many studies have been conducted on the biologically active constituents of whole parts of M. bontioides. However, the endophytes of M. bontioides have not been intensively investigated. A new chlorine-containing isocoumarin, named dichlorodiaportinol A (1) was isolated from the endophytic fungus Trichoderma sp. 09 isolated from the root of M. bontioides. Its cytotoxic activity against human breast cancer (MCF-7) and human liver cancer (HepG2) cell lines was evaluated. Materials and Methods: Different open silica gel column chromatographic techniques with different solvent systems were used for the separation of the constituents of the ethyl acetate extract of the culture broth of the endophytic fungus Trichoderma sp. 09. The structure of compound one was identified by analysis of spectroscopic data [one-dimensional (1D), two-dimensional (2D)-nuclear magnetic resonance (NMR), ultraviolet (UV), infrared (IR) and Mass spectrometry (MS)]. 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay method was used for the evaluation of cytotoxic activity of compound one against MCF-7 and HepG2 cell lines. Results: Compound one was identified as 3-(3,3-dichloro-2,3-dihydroxy-propyl)-8-hydroxy-6- methoxy-isochromen-1-one. It inhibited MCF-7 and HepG2 cell lines, with half maximal inhibitory concentration (IC 50 ) values of 17.8 and 39.6 μg/mL, respectively. Conclusions: Compound one is a new chlorine-containing isocoumarin with moderate cytotoxic activity against MCF-7 and HepG2 cell lines. Thus, endophytes of M. bontioides are worthy of consideration for the development and research of antitumor agents.
  1 1,645 24
Chemical composition of the volatile oil from Zanthoxylum avicennae and antimicrobial activities and cytotoxicity
Yin Lin, Wei Han, Wei-chen Ge, Ke Yuan
January 2014, 10(37):164-170
DOI:10.4103/0973-1296.127369  PMID:24914299
Background: Through literature retrieval, there has been no report on the research of the chemical components in Zanthoxylum avicennae (Lam.) DC. This paper extracted and determined the chemical components of the volatile oil in Z. avicennae, and at the same time, measured and evaluated the bioactivity of the volatile oil in Z. avicennae. Materials and Methods: We extract the volatile oil in Z. avicennae by steam distillation method, determined the chemical composition of the volatile oil by GC-MS coupling technique, and adopt the peak area normalization method to measured the relative percentage of each chemical composition in the volatile oil. Meanwhile, we use the Lethal-to-prawn larva bioactivity experiment to screen the cytotoxicity activities of the volatile oil in Z. avicennae, and using the slanting test-tube experiment to determine and evaluate its antibacterial activities in vitro for the eight kinds of plant pathogenic fungi in the volatile oil of the Z. avicennae. Results: The results show that 68 kinds of compounds are determined from the volatile oil of Z. avicennae. The determined part takes up 97.89% of the total peak area. The main ingredients in the volatile oil of Z. avicennae are sesquiterpenoids and monoterpene. The test results show that the volatile oil in Z. avicennae has strong antibacterial activities and cytotoxicity, with the strongest antibacterial activity against the Rhizoctonia solani AG1-1A. Conclusion: This research results will provide reference data for understanding the chemical composition of the volatile oil in the aromatic plant of Z. avicennae and its bioactivity, and for its further development and application.
  1 1,467 21
Comparison of pharmacokinetic behavior of two iridoid glycosides in rat plasma after oral administration of crude Cornus officinals and its jiuzhipin by high performance liquid chromatography triple quadrupole mass spectrometry combined with multiple reactions monitoring mode
Xiaocheng Chen, Gang Cao, Jianping Jiang
January 2014, 10(37):115-121
DOI:10.4103/0973-1296.127358  PMID:24914290
Objective: The present study examined the pharmacokinetic profiles of two iridoid glycosides named morroniside and loganin in rat plasma after oral administration of crude and processed Cornus officinals. Materials and Methods: A rapid, selective and specific high-performance liquid chromatography/electrospray ionization tandem mass spectrometry with multiple reactions monitoring mode was developed to simultaneously investigate the pharmacokinetic profiles of morroniside and loganin in rat plasma after oral administration of crude C. officinals and its jiuzhipin. Results: The morroniside and loganin in crude and processed C. officinals could be simultaneously determined within 7.4 min. Linear calibration curves were obtained over the concentration ranges of 45.45-4800 ng/mL for all the analytes. The intra-and inter-day precisions relative standard deviation was lesser than 2.84% and 4.12%, respectively. Conclusion: The pharmacokinetic parameters of two iridoid glucosides were also compared systematically between crude and processed C. officinals. This paper provides the theoretical proofs for further explaining the processing mechanism of Traditional Chinese Medicines.
  1 1,353 17
Characteristics and kinetics of catalpol degradation and the effect of its degradation products on free radical scavenging
Guo-dong Wei, Xue-sen Wen
January 2014, 10(37):122-129
DOI:10.4103/0973-1296.127360  PMID:24914291
Background: The dried and steamed roots of Rehmannia glutinosa have different pharmacological functions and indications. Catalpol, the main active component of the dried root, was found to be entirely degraded together with amino acids and some oligosaccharides during preparation of the steamed root. Its degradation may contribute to the differences between dried and steamed roots. Objective: To reveal the characteristics and kinetics of catalpol degradation, and evaluate its influence on the antioxidant properties of steamed Rehmannia roots. Materials and Methods: Purified catalpol was heated under different pH and temperature values for different times, alone or with sugars or amino acids. Catalpol concentration was determined by high-performance liquid chromatography. Browning was expressed by the absorbance at 420 nm (A 420 ), and antioxidation was displayed by 2,2-diphenyl-1-picrylhydrazyl free radical scavenging ability (SA DPPH ). Activation energy was calculated using Arrhenius plotting. Results: Catalpol was stable in neutral conditions and sensitive to acidic pH under high temperatures. Sugars had no influence on catalpol degradation; however, most amino acids, except for proline, could promote the degradation, and were associated with an increase in A 420 and SA DPPH values. These changes were proved to be mainly related with catalpol aglycone and were dependent on the presence of amino acids. Catalpol degradation was found to obey first-order kinetics. The activation energies were 81.7, 88.8 and 98.7 kJ/mol at pH 4.0, 5.0, and 6.0 respectively, and 70.7 kJ/mol at pH 4.0 value and in the presence of glycine. Conclusions: Catalpol degradation, especially, in the presence of amino acids can substantially boost antioxidant properties of the products; therefore, the traditional method for processing Rehmannia root seems rather apt.
  1 1,419 19
Composition of the essential oil of Cynanchum mongolicum (Asclepiadaceae) and insecticidal activities against Aphis glycines (Hemiptera: Aphidiae)
Wang Yang, An Zhao, Zhen Congai, Liu Qizhi, Shi Wangpeng
January 2014, 10(37):130-134
DOI:10.4103/0973-1296.127362  PMID:24914292
Background: Farmers have applied Cynanchum mongolicum (Maxim) to control crop pests. The aim of this study was to analyze composition of essential oil from C. mongolicum, evaluate insecticidal activities against Aphis glycines, and lethal doses. Materials and Methods: Essential oil from C. mongolicum was efficiently extracted by steam distillation. The main components of the oil were analyzed with a gas chromatography/mass spectrometry (GC/MS) system, and the insecticidal activity of the essential oil on soybean aphids Aphis glycines was tested using a variety of methods. Results: The components of the essential oil from C. mongolicum mainly included terpenes and ester compounds, of which (Z)-3-Hexen-1-ol acetate, cis-3-hexenyl isovalerate, α-farnesene, and β-caryophyllene accounted for 15.8, 10.4, 8.4, and 5.5%, respectively. With 1- and 2-day exposure, the essential oil showed pronounced contact toxicity (median lethal concentration (LC 50 ) =37.8 and 38.4 μL/mL, respectively), weak fumigant toxicity (LC 50 = 139.7 and 139.9 μL/L, respectively). The essential oil showed strong deterrent activity on soybean aphids in 2 and 4 h. Conclusion: The essential oil of C. mongolicum contained insecticidal components and possessed contact toxicity and deterrent activity to A. glycines.
  1 2,126 14
Crude ethyl acetate extract of marine microalga, Chaetoceros calcitrans, induces Apoptosis in MDA-MB-231 breast cancer cells
Su Hua Goh, Noorjahan Banu Mohamed Alitheen, Fatimah Md Yusoff, Swee Keong Yap, Su Peng Loh
January-March 2014, 10(37):1-8
DOI:10.4103/0973-1296.126650  PMID:24696543
Background: Marine brown diatom Chaetoceros calcitrans and green microalga Nannochloropsis oculata are beneficial materials for various applications in the food, nutraceutical, pharmaceutical and cosmeceutical industries. Objective: This study investigated cytotoxicity of different crude solvent extracts from C. calcitrans and N. oculata against various cancer cell lines. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was carried out to screen the cytotoxic effects of hexane (Hex), dichloromethane (DCM), ethyl acetate, and methanol extract from C. calcitrans and N. oculata toward various cancer cell lines. Flow cytometry cell cycle was used to determine the cell cycle arrest while the mode of cell death was investigated through acridine orange/propidium iodide (AOPI) staining, Annexin V-Fluorescein Isothiocyanate (FITC) and Terminal deoxynucleotidyl transferase-mediated d-UTP Nick End Labeling (TUNEL) assays. Expression profile of apoptotic and proliferative-related genes was then determined using the multiplex gene expression profiler (GeXP). Results: Crude ethyl acetate (CEA) extract of C. calcitrans inhibited growth of MDA-MB-231 cells, with IC 50 of 60 μg/mL after 72 h of treatment. Further studies were conducted to determine the mode of cell death at various concentrations of this extract: 30, 60 and 120 μg/mL. The mode of cell death was mainly apoptosis as shown through apoptosis determination test. The expression data from GeXP showed that caspase-4 was upregulated while B-cell leukemia/lymphoma 2(Bcl-2) was down regulated. Thus, caspase-4 induction endoplasmic reticulum death pathway is believed to be one of the mechanisms underlying the induction of apoptosis while Bcl-2 induced S and G2/M cell cycle phase arrest in MDA-MB-231 cells. Conclusion: CEA extract of C. calcitrans showed the highest cytotoxicity on MDA-MB-231 via apoptosis.
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Assessment of potency of PC-complexed Ocimum sanctum methanol extract in embryonated eggs against Influenza virus (H1N1)
Priyanka Jadhav, Hingorani Lal, Nilima Kshirsagar
January 2014, 10(37):86-91
DOI:10.4103/0973-1296.127352  PMID:24914314
Background: Despite of new vaccines, the threat of influenza infection persists. In addition, availability, cost, duration of protection rendered and effectiveness of vaccines additional to the need of effective drug therapy makes influenza a challenge, which the globe faces. Traditionally used herbs and their decoctions are used for ages to cure symptoms similar to influenza. Tulsi or Ocimum sanctum is one of these major herbs used for influenza-like disease treatment. We attempted to explore a new methodology for assessing phosphatidyl choline (PC)-complexed O. sanctum methanol extract in embryonated vaccine quality eggs model. Materials and Methods: The PC-complexed O. sanctum methanol extract was prepared and standardized using High-Performance Liquid Chromatography (HPLC). (Data not provided here) Nine to 11 days embryonated eggs were inoculated with the virus and drug mixture and then harvested to perform a hemagglutination (HA) test on the allantoic fluid. The experiments were performed at three different concentrations of ursolic acid with various virus concentration and dose levels of drugs. The HA titer was calculated from all experiments and observed for any inhibition of virus. Results: In initial experiments, matrix method for drug and virus concentration was employed. It was observed that the drug exhibited some response for 3log EID 50 (egg infective dose) in few samples at 1:2 HA titer, but no response was observed at 4log EID 50 . In subsequent experiment, all the virus titers from 7log EID 50 to 2log EID 50 demonstrated positive HA titer of 1:64. However, the drug failed to exhibit any significant inhibition at any level of demonstrable virus titer. At all the concentrations, O. sanctum extracts were found to be safe. Conclusion: The embryonated egg model may be utilized further to screen other drugs, which possess direct inhibitory properties like neuraminidase inhibition, and O. sanctum does not inhibit the influenza virus in this model at the given concentration.
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Compounds isolated from Ageratum houstonianum inhibit the activity of matrix metalloproteinases (MMP-2 and MMP-9): An oncoinformatics study
Anupriya Verma, Syed Mohd Danish Rizvi, Sibhghatulla Shaikh, Mohd Afaque Ansari, Shazi Shakil, Fauzia Ghazal, Mohd Haris Siddiqui, Mohd Haneef, Ajijur Rehman
January-March 2014, 10(37):18-26
DOI:10.4103/0973-1296.126653  PMID:24695379
Background: In osteosarcoma tissue, both MMP-2 and MMP-9 are over expressed compared to their expression in non-affected stromal tissue. Hence, gelatinases are attractive targets for anti-osteosarcoma drugs. Objective: To study the inhibitory activity of compounds isolated from Ageratum houstonianum against MMP-2 and MMP-9 by in-silico approach. Material and Methods: We performed docking study using 'Autodock 4.2' between 1,2-benzenedicarboxylic acid-bis (2-ethylhexyl) ester; squalene; 3,5-bis (1,1-dimethylethyl) phenol; pentamethyl tetrahydro-5H-chromene; (1, 4-cyclohexylphenyl) ethanone and 6-vinyl-7-methoxy-2,2-dimethylchromene with MMP-2 and MMP-9. Results: Among all six compounds isolated from Ageratum houstonianum, (1, 4-cyclohexylphenyl) ethanone showed the maximum potential as a putative inhibitor of both MMP-2 and MMP-9 enzymes with reference to ΔG (−7.95 and −8.2 kcal/mol, respectively) and Ki (1.48 and 0.98 μM, respectively) values. Total intermolecular energy of docking for (1, 4-cyclohexylphenyl) ethanone-MMP catalytic domain-interaction was found to be −8.55 kcal/mol for MMP-2 and −9.21 kcal/mol for MMP-9. Conclusion: This study explores molecular interactions between human MMPs (MMP-2 and MMP-9) and six natural compounds. This study predicts that (1,4-cyclohexylphenyl) ethanone is a more efficient inhibitor of human MMP-2 and MMP-9 enzymes compared to the other natural compounds used in this study with reference to Ki and ΔG values.
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High-performance liquid chromatography determination and pharmacokinetics of coumarin compounds after oral administration of Samul-Tang to rats
Youn-Hwan Hwang, Won-Kyung Cho, Doorye Jang, Jeong-Ho Ha, Jin Yeul Ma
January-March 2014, 10(37):34-39
DOI:10.4103/0973-1296.126656  PMID:24696544
Background: Samul-tang has been traditionally used for the treatment of cardiovascular, gynecologic, cutaneous, and chronic inflammation disorders. Although coumarin compounds do have various pharmacological activities and the same may be present in Samul-tang, however there is little information about it. Objective: A simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for the determination of nodakenin, nodakenetin, decursin, decursinol, and decursinol angelate in rat plasma. To obtain a better understanding for pharmacological properties of Samul-tang, pharmacokinetic study of coumarin compounds was performed after oral administration of Samul-tang in rats. Materials and Methods: Chromatographic separation of the analytes was successfully achieved on a Phenomenex Luna C 18 column (4.6 mm×250 mm, 5 μm) using a mobile phase composed of acetonitrile water with a gradient elution at a flow rate of 1 mL/min. Noncompartmental analysis was performed. Results: Calibration curves for all analytes had good linearity (r 2 <0.999) in a wide linear range. The lower limit of quantification (LLOQ) ranged from 0.05 to 0.1 μg/mL. The variation of intra- and interday assay was less than 15%. Nodakenin, nodakenetin, and decursinol were determined in rat plasma after oral administration of Samul-tang. Conclusion: This developed and validated HPLC method was successfully applied to the pharmacokinetic study of three coumarin compounds in rats, given as a single oral administration of Samul-tang. These pharmacokinetic data of the nodakenin, nodakenetin, and decursinol could offer a new point of view to evaluate the pharmacological effects of Samul-tang.
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Fraction and chemical analysis of antioxidant active polysaccharide isolated from flue-cured tobacco leaves
Chunping Xu, Chenchen Yang, Duobin Mao
January-March 2014, 10(37):66-69
DOI:10.4103/0973-1296.126664  PMID:24696548
Background: The metabolic compounds from tobacco were investigated to posses various biological activities, such as antioxidant, antimicrobial and neuroprotective activities. Materials and Methods: Polysaccharides were isolated from the ultrasound-assisted extraction of flue-cured tobacco with hot water. The purified polysaccharides were analyzed by GC-MS, FT-IR, and TGA (Thermogravimetric analysis). The antioxidant activities of purified polysaccharides were evaluated in vitro. Results: Two polysaccharides (Fr-I and Fr-II) were isolated and purified. The analysis of monosaccharide composition in the polysaccharide by GC/MS revealed that Fr-I and Fr-II were heteropolysaccharides. Fr-I consisted of D-mannose and galactose, ribose and arabinose, while Fr-II was mainly composed of glucose, allose, galactose and mannose. Thermo gravimetric analysis (TGA) indicated that the degradation temperature (Td) of the Fr-I (241 °C) was higher than that of Fr-II (220 °C). Both samples showed inhibitory effects on OH (hydroxyl) and DPPH (2, 2-diphenyl-1-picryl-hydrazyl-hydrate) radical in a concentration-dependent manner. Comparing Fr-I with Fr-II, the latter has a strong scavenging ability. Conclusion: Both polysaccharide fractions showed significant antioxidant effects. Various factors influenced the antioxidant activity of polysaccharides.
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Identification of sulfur fumed Pinelliae Rhizoma using an electronic nose
Xia Zhou, Jun Wan, Liang Chu, Wengang Liu, Yafeng Jing, Chunjie Wu
January 2014, 10(37):135-140
DOI:10.4103/0973-1296.127363  PMID:24914293
Background: Pinelliae Rhizoma is a commonly used Chinese herb which will change brown during the natural drying process. However, sulfur fumed Pinelliae Rhizoma will get a better appearance than naturally dried one. Sulfur fumed Pinelliae Rhizoma is potentially toxical due to sulfur dioxide and sulfites formed during the fuming procedures. The odor components in sulfur fumed Pinelliae Rhizoma is complex. At present, there is no analytical method available to determine sulfur fumed Pinelliae Rhizoma simply and rapidly. To ensure medication safety, it is highly desirable to have an effective and simple method to identify sulfur fumed Pinelliae Rhizoma. Materials and Methods: This paper presents a novel approach using an electronic nose based on metal oxide sensors to identify whether Pinelliae Rhizoma was fumed with sulfur, and to predict the fuming degree of Pinelliae Rhizoma. Multivariate statistical methods such as principal components analysis (PCA), discriminant factorial analysis (DFA) and partial least squares (PLS) were used for data analyzing and identification. The use of the electronic nose to discriminate between different fuming degrees Pinelliae Rhizoma and naturally dried Pinelliae Rhizoma was demonstrated. Results: The electronic nose was also successfully applied to identify unknown samples including sulfur fumed samples and naturally dried samples, high recognition value was obtained. Quantitative analysis of fuming degree of Pinelliae Rhizoma was also demonstrated. The method developed is simple and fast, which provides a new quality control method of Chinese herbs from the aspect of odor. Conclusion: It has shown that this electronic nose based metal oxide sensor is sensitive to sulfur and sulfides. We suggest that it can serve as a supportive method to detect residual sulfur and sulfides.
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An enzoinformatics study targeting polo-like kinases-1 enzyme: Comparative assessment of anticancer potential of compounds isolated from leaves of Ageratum houstonianum
Syed Mohd Danish Rizvi, Shazi Shakil, Mohd Zeeshan, Mohd Sajid Khan, Sibhghatulla Shaikh, Deboshree Biswas, Adnan Ahmad, Mohammad Amjad Kamal
January 2014, 10(37):14-21
DOI:10.4103/0973-1296.127333  PMID:24914294
Natural products from plant sources, embracing inherently ample structural diversity than synthetic ones are the major sources of anticancer agents and will constantly play as protagonists for discovering new drugs. Polo-like kinases (PLKs) play a leading role in the ordered execution of mitotic events and 4 mammalian PLK family members have been identified. PLK1 is an attractive target for anticancer drugs in mammalian cells, among the four members of PLKs. The present study expresses the molecular interaction of compounds (1,2-Benzenedicarboxylic acid bis (2 ethylhexyl) ester, squalene, 3,5-bis (1,1-dimethylethyl) phenol, Pentamethyl tetrahydro-5H-chromene, (1,4-Cyclohexylphenyl) ethanone and 6-Vinyl-7-methoxy-2,2-dimethylchromene) isolated from methanolic extract of leaves of Ageratum houstonianum with PLK1 enzyme. Docking between PLK1 and each of these compounds (separately) was performed using "Auto dock 4.2." (1,4-Cyclohexylphenyl) ethanone showed the maximum potential as a promising inhibitor of PLK1 enzyme with reference to ∆G (−6.84 kcal/mol) and Ki (9.77 μM) values. This was sequentially followed by Pentamethyl tetrahydro-5H-chromene (∆G = −6.60 kcal/mol; Ki = 14.58 μM), squalene (∆G = −6.17 kcal/mol; Ki = 30.12 μM), 6-Vinyl-7-methoxy-2,2-dimethylchromene (∆G = −5.91 kcal/mol; Ki = 46.68 μM), 3, 5-bis (1,1-dimethylethyl) phenol (∆G = −5.70 kcal/mol; Ki = 66.68 μM) and 1,2-Benzenedicarboxylic acid bis (2 ethylhexyl) ester (∆G = −5.58 kcal/mol; Ki = 80.80 μM). These results suggest that (1,4-Cyclohexylphenyl) ethanone might be a potent PLK1 inhibitor. Further, in vitro and in vivo rumination are warranted to validate the anticancer potential of (1,4-Cyclohexylphenyl) ethanone.
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Ultrasound-assisted extraction of total flavonoids from Aconitum gymnandrum
Ling-Li Zheng, Dan Wang, Yuan-Yuan Li, Hong-Yan Peng, Ming-Yong Yuan, Feng Gao
January 2014, 10(37):141-146
DOI:10.4103/0973-1296.127364  PMID:24914295
Background: Aconitum gymnandrum is a Chinese traditional herb used as carminative and analgesic. In this study, A. gymnandrum was used as an experimental matrix. Materials and Methods: Optimized ultrasonic extraction technology of total flavonoids from the A. gymnandrum Maxim was studied by using the methodology of single factor and orthogonal design to study the effects of operation conditions, such as ethanol content, ultrasonic wave power, temperature, ultrasonic wave radiation time, and the ratio of sample weight to solvent volume. Result: Through the orthogonal experiment, the optimal extraction conditions were determined as follows: Ultrasonic power 100 W, ultrasonic temperature 45°C, 60% ethyl alcohol, extraction time 30 min, and solid-liquid ratio 1:20. Conclusion: Under the optimum parameters, the extraction ratio of total flavonoids from the A. gymnandrum Maxim is about 1.278%.
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Increased acetylcholine esterase activity produced by the administration of an aqueous extract of the seed kernel of Thevetia peruviana and its role on acute and subchronic intoxication in mice
Rubén Marroquín-Segura, Ricardo Calvillo-Esparza, José Luis Alfredo Mora-Guevara, José Horacio Tovalín-Ahumada, Abigail Aguilar-Contreras, Vicente Jesús Hernández-Abad
January 2014, 10(37):171-175
DOI:10.4103/0973-1296.127370  PMID:24914300
Background: The real mechanism for Thevetia peruviana poisoning remains unclear. Cholinergic activity is important for cardiac function regulation, however, the effect of T. peruviana on cholinergic activity is not well-known. Objective: To study the effect of the acute administration of an aqueous extract of the seed kernel of T. peruviana on the acetylcholine esterase (AChE) activity in CD1 mice as well its implications in the sub-chronic toxicity of the extract. Materials and Methods: A dose of 100 mg/kg of the extract was administered to CD1 mice and after 7 days, serum was obtained for ceruloplasmin (CP) quantitation and liver function tests. Another group of mice received a 50 mg/kg dose of the extract 3 times within 1 h time interval and AChE activity was determined for those animals. Heart tissue histological preparation was obtained from a group of mice that received a daily 50 mg/kg dose of the extract by a 30-days period. Results: CP levels for the treated group were higher than those for the control group (Student's t-test, P ≤ 0.001). AChE activity in the treated group was significantly higher than the control group (Tukey test, control vs. T. peruviana, P ≤ 0.001). Heart tissue histological preparations showed leukocyte infiltrates and necrotic areas, consistent with infarcts. Conclusion: The increased levels of AChE and the hearth tissue infiltrative lesions induced by the aqueous seed kernel extract of T. peruviana explains in part the poisoning caused by this plant, which can be related to an inflammatory process.
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Simultaneous determination of five major compounds in the traditional medicine Pyeongwee-San by high performance liquid chromatography-diode array detection and liquid chromatography-mass spectrometry/mass spectrometry
Bohyoung Lee, Jin Bae Weon, Bo-Ra Yun, Jiwoo Lee, Min Rye Eom, Choong Je Ma
January 2014, 10(37):22-29
DOI:10.4103/0973-1296.127335  PMID:24914305
Background: Pyeongwee-San (PWS) has been widely used for treating acute gastritis, chronic, and gastritis. Objective: In this paper, simultaneous determination of five compounds (naringin, hesperidin, glycyrrhizin, atractylenolide III, and magnolol) from traditional medicine PWS using the high performance liquid chromatography (HPLC) was established for quality control. Materials and Methods: Optimum separations were obtained with a SHISEIDO C18 reverse-phase column by gradient elution with 0.1% Trifluoroacetic acid (TFA) water-acetonitrile as the mobile phase. The flow rate was 1 mL/min and detection wavelength was set at 205 nm and 250 nm. Validation of the analytical method was evaluated by linearity, precision, and accuracy test. Results: The calibration curves were linear over the established range with R 2 > 0.9978. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.09 to 0.43 and 0.27 to 1.29 μg/mL. The method exhibited intra-day and inter-day precision range between 0.01-1.86% and 0.04-0.35% respectively. The recoveries of five compounds in PWS were in the range between 93.18-106.40%, and 0.20-1.51%. The application of this method was identified through the successful analysis of five compounds in 12 batches of PWS. In addition, identification of five compounds was confirmed by a liquid chromatography method and mass spectrometry. Conclusion: The HPLC method was could be accomplished to the quality control and stable experiment for the preparations consisted of five major compounds.
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