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   2016| January-March  | Volume 12 | Issue 45  
    Online since February 10, 2016

 
 
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ORIGINAL ARTICLES
Comparative pharmacokinetics of ginsenoside Rg3 and ginsenoside Rh2 after oral administration of ginsenoside Rg3 in normal and walker 256 tumor-bearing rats
He Fan, Sun Xiao-ling, Su Yaliu, Lu Ming-ming, Feng Xue, Meng Xian-sheng, Fu Li
January-March 2016, 12(45):21-24
DOI:10.4103/0973-1296.176014  PMID:27019557
Background: Ginseng is Chinese traditional herbal medicine, and the ginsenoside Rg 3 is the main bioactive ingredient for the anti-tumor effect. However, there is no study on pharmacokinetics (PKs) of ginsenoside Rg 3 and its main metabolite after oral ginsenoside Rg 3 in tumor-bearing plasma. The aim of this study was to investigate the PK profiles of ginsenoside Rg 3 and ginsenoside Rh 2 after oral administration of pure ginsenoside Rg 3 were administered, and compare the difference of the PK profiles between normal and Walker 256 tumor-bearing rats. Materials and Methods: The concentrations of two ginsenosides in plasma were determined by using a simple and rapid high-performance liquid chromatography. All the rats were divided randomly into two groups (Walker 256 tumor-bearing and normal groups). Each group received oral administration of 50 mg/kg ginsenoside Rg 3 . Results: The results showed that ginsenoside Rh 2, possibly as a glycosylation hydrolysis product of ginsenoside Rg 3, were found in plasma after oral administration of ginsenoside Rg 3 to rats. Ginsenoside Rg 3 had shown better absorption than ginsenoside Rh 2, whether the oral administration of ginsenoside Rg 3 , normal rats showed better absorption than tumor-bearing rats. Discussion and Conclusion: The PKs properties of the ginsenoside Rg 3 and ginsenoside Rh 2 differed between tumor-bearing rats and normal rats, including area under the plasma level/time curve and concentration maximum (P < 0.05).
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Chemical discrimination of cortex Phellodendri amurensis and cortex Phellodendri chinensis by multivariate analysis approach
Hui Sun, Huiyu Wang, Aihua Zhang, Guangli Yan, Ying Han, Yuan Li, Xiuhong Wu, Xiangcai Meng, Xijun Wang
January-March 2016, 12(45):41-49
DOI:10.4103/0973-1296.176023  PMID:27018001
Background: As herbal medicines have an important position in health care systems worldwide, their current assessment, and quality control are a major bottleneck. Cortex Phellodendri chinensis (CPC) and Cortex Phellodendri amurensis (CPA) are widely used in China, however, how to identify species of CPA and CPC has become urgent. Materials and Methods: In this study, multivariate analysis approach was performed to the investigation of chemical discrimination of CPA and CPC. Results: Principal component analysis showed that two herbs could be separated clearly. The chemical markers such as berberine, palmatine, phellodendrine, magnoflorine, obacunone, and obaculactone were identified through the orthogonal partial least squared discriminant analysis, and were identified tentatively by the accurate mass of quadruple-time-of-flight mass spectrometry. A total of 29 components can be used as the chemical markers for discrimination of CPA and CPC. Of them, phellodenrine is significantly higher in CPC than that of CPA, whereas obacunone and obaculactone are significantly higher in CPA than that of CPC. Conclusion: The present study proves that multivariate analysis approach based chemical analysis greatly contributes to the investigation of CPA and CPC, and showed that the identified chemical markers as a whole should be used to discriminate the two herbal medicines, and simultaneously the results also provided chemical information for their quality assessment.
  3 1,727 24
Theanine and Caffeine content of infusions prepared from commercial tea samples
Klára Boros, Nikoletta Jedlinszki, Dezső Csupor
January-March 2016, 12(45):75-79
DOI:10.4103/0973-1296.176061  PMID:27019564
Background: Caffeine and L-theanine are pharmacologically important constituents of tea, especially due to their effects on the central nervous system. The effects of these two compounds are opposite: While caffeine is a well-known stimulant, theanine has a relaxing effect. Tea processing may influence the caffeine and theanine content of tea leaves. Objective: The aim of our work was to quantify these constituents from a set of commercial products to reveal the possible correlations of caffeine and theanine content and processing methods. Materials and Methods: Theanine and caffeine contents of 37 commercial white, green, oolong, black, and pu-erh tea samples were quantified by high-performance liquid chromatography-diode array detector. Results: The mean L-theanine content of white, green, oolong, and black teas were 6.26, 6.56, 6.09, and 5.13 mg/g, respectively. The same values for caffeine content were 16.79, 16.28, 19.31, and 17.73 mg/g. Conclusion: Though the effect of processing on theanine content was evident, quantification for these analytes does not seem to be a good criterion to discriminate the different types of tea. Caffeine content provided no information on the effect of processing, and the theanine content of the samples was rather variable, independently from the type of the tea. The quantitative analysis of caffeine and theanine is essential to assess the stimulating effect of the tea, however, for chemical profiling further secondary metabolites have to be determined.
  3 2,854 22
Comparation of hypolipidemic and antioxidant effects of aqueous and ethanol extracts of Crataegus pinnatifida fruit in high-fat emulsion-induced hyperlipidemia rats
Feng Shao, Lifei Gu, Huijuan Chen, Ronghua Liu, Huilian Huang, Gang Ren
January-March 2016, 12(45):64-69
DOI:10.4103/0973-1296.176049  PMID:27019563
Background: Hawthorn (Crataegus pinnatifida) is a Chinese medicinal plant traditionally used in the treatment of hyperlipidemia. Recently, studies indicated free radical scavenging was one of the major pathways to alleviate hyperlipidemia. Moreover, hawthorn fruit is a rich source of phenols, which quench free radical and attenuate hyperlipidemia. However, the phenols vary with processing methods, especially solvent type. Objective: Our aim was to compare hypolipidemic and antioxidant effects of aqueous and ethanol extracts of hawthorn fruit in hyperlipidemia rats. Materials and Methods: After a 4-week treatment of high-fat emulsion, lipid profile levels and antioxidant levels of two extracts were determined using commercial analysis. Total phenols content in the extract of hawthorn fruit was determined colorimetrically by the Folin-Ciocalteu method. Results: Both ethanol and aqueous extracts of hawthorn fruit possessed hypolipidemic and antioxidant activities. Simultaneously, stronger activities were observed in ethanol extract. Besides, total phenols content in ethanol extract from the same quality of hawthorn fruit was 3.9 times more than that in aqueous extract. Conclusion: The obvious difference of hypolipidemic and antioxidant effects between ethanol extract and aqueous extract of hawthorn fruit was probably due to the presence of total phenols content, under the influence of extraction solvent.
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Protective mechanisms of thymoquinone on methotrexate-induced intestinal toxicity in rats
Azza A El-Sheikh, Mohamed A Morsy, Azza H Hamouda
January 2016, 12(45):76-81
DOI:10.4103/0973-1296.176106  PMID:27041864
Background: Intestinal toxicity is a serious side effect in methotrexate (MTX) chemotherapy. Objective: To investigate the mechanisms by which the anticancer drug MTX-induced intestinal damage could be prevented by thymoquinone (TQ), an active ingredient of Nigella sativa. Materials and Methods: TQ was given orally for 10 days, and MTX toxicity was induced at the end of day 3 of the experiment, with or without TQ pretreatment. Results: MTX caused intestinal damage, represented by distortion in normal intestinal histological structure, with significant oxidative stress, exhibited as decrease in reduced glutathione concentration and catalase activity, along with significant increase in malondialdehyde level compared to control group. MTX also caused nitrosative stress evident by increased intestinal nitric oxide (NO) level, with up-regulation of inducible NO synthase expression shown in immunohistochemical staining. Furthermore, MTX caused inflammatory effects as evident by up-regulation of intestinal necrosis factor-kappa beta and cyclooxygenase-2 expressions, which were confirmed by increased intestinal tumor necrosis factor-alpha level via enzyme-linked immunosorbent assay. Moreover, MTX caused apoptotic effect, as it up-regulated intestinal caspase 3 expression. Concomitant TQ significantly reversed the MTX-induced intestinal toxic effects by reversing intestinal microscopic damage, as well as significantly improving oxidative/nitrosative stress, inflammatory and apoptotic markers tested compared to MTX alone. Conclusion: TQ may possess beneficial intestinal protective effects as an adjuvant co-drug against MTX intestinal toxicity during cancer chemotherapy. TQ protection is conferred via antioxidant, anti-nitrosative, anti-inflammatory, and anti-apoptotic mechanisms.
  3 1,621 19
Naringin and rutin alleviates episodic memory deficits in two differentially challenged object recognition tasks
Grandhi Venkata Ramalingayya, Madhavan Nampoothiri, Pawan G Nayak, Anoop Kishore, Rekha R Shenoy, Chamallamudi Mallikarjuna Rao, Krishnadas Nandakumar
January 2016, 12(45):63-70
DOI:10.4103/0973-1296.176104  PMID:27041861
Background: Cognitive decline or dementia is a debilitating problem of neurological disorders such as Alzheimer's and Parkinson's disease, including special conditions like chemobrain. Dietary flavonoids proved to be efficacious in delaying the incidence of neurodegenerative diseases. Two such flavonoids, naringin (NAR) and rutin (RUT) were reported to have neuroprotective potential with beneficial effects on spatial and emotional memories in particular. However, the efficacy of these flavonoids is poorly understood on episodic memory, which comprises an important form of autobiographical memory.Objective: This study objective is to evaluate NAR and RUT to reverse time-delay-induced long-term and scopolamine-induced short-term episodic memory deficits in Wistar rats. Materials and Methods: We have evaluated both short-term and long-term episodic memory forms using novel object recognition task. Open field paradigm was used to assess locomotor activity for any confounding influence on memory assessment. Donepezil was used as positive control and was effective in both models at 1 mg/kg, i.p. Results: Animals treated with NAR and RUT at 50 and 100 mg/kg, p.o. spent significantly more time exploring novel object compared to familiar one, whereas control animals spent almost equal time with both objects in choice trial. NAR and RUT dose-dependently increased recognition and discriminative indices in time-induced long-term as well as scopolamine-induced short-term episodic memory deficit models without interfering with the locomotor activity. Conclusion:We conclude that, NAR and RUT averted both short- and long-term episodic memory deficits in Wistar rats, which may be potential interventions for neurodegenerative diseases as well as chemobrain condition.
  2 1,524 19
Liquid chromatography-diode array detector-electrospray mass spectrometry and principal components analyses of raw and processed moutan cortex
Xian-Mei Deng, Jiang-Yong Yu, Meng-Jin Ding, Ming Zhao, Xing-Yang Xue, Chun-Tao Che, Shu-Mei Wang, Bin Zhao, Jiang Meng
January-March 2016, 12(45):50-56
DOI:10.4103/0973-1296.176046  PMID:27019561
Background: Raw Moutan Cortex (RMC), derived from the root bark of Paeonia suffruticosa, and Processed Moutan Cortex (PMC) is obtained from RMC by undergoing a stir-frying process. Both of them are indicated for different pharmacodynamic action in traditional Chinese medicine, and they have been used in China and other Asian countries for thousands of years. Objective: To establish a method to study the RMC and PMC, revealing their different chemical composition by fingerprint, qualitative, and quantitative ways. Materials and Methods: High-performance liquid chromatography coupled with diode array detector and electrospray mass spectrometry (HPLC-DAD-ESIMS) were used for the analysis. Therefore, the analytes were separated on an Ultimate TM XB-C18 analytical column (250 mm × 4.6 mm, 5.0 μm) with a gradient elution program by a mobile phase consisting of acetonitrile and 0.1% (v/v) formic acid water solution. The flow rate, injection volume, detection wavelength, and column temperature were set at 1.0 mL/min, 10 μL, 254 nm, and 30°C, respectively. Besides, principal components analysis and the test of significance were applied in data analysis. Results: The results clearly showed a significant difference among RMC and PMC, indicating the significant changes in their chemical compositions before and after the stir-frying process. Conclusion: The HPLC-DAD-ESIMS coupled with chemometrics analysis could be used for comprehensive quality evaluation of raw and processed Moutan Cortex.
  2 1,622 25
Nuclear factor kappa B activation and peroxisome proliferator-activated receptor transactivational effects of chemical components of the roots of Polygonum multiflorum
Ya Nan Sun, Wei Li, Seok Bean Song, Xi Tao Yan, Seo Young Yang, Young Ho Kim
January-March 2016, 12(45):31-35
DOI:10.4103/0973-1296.176019  PMID:27019559
Background: Polygonum multiflorum is well-known as "Heshouwu" in traditional Chinese herbal medicine. In Northeast Asia, it is often used as a tonic to prevent premature aging of the kidney and liver, tendons, and bones and strengthening of the lower back and knees. Objective: To research the anti-inflammatory activities of components from P. multiflorum. Materials and Methods: The compounds were isolated by a combination of silica gel and YMC R-18 column chromatography, and their structures were identified by analysis of spectroscopic data (1D, 2D-nuclear magnetic resonance, and mass spectrometry). The anti-inflammatory activities of the isolated compounds 1−15 were evaluated by luciferase reporter gene assays. Results: Fifteen compounds (1-15) were isolated from the roots of P. multiflorum. Compounds 1−5 and 14−15 significantly inhibited tumor necrosis factor-α-induced nuclear factor kappa B-luciferase activity, with IC 50 values of 24.16-37.56 μM. Compounds 1−5 also greatly enhanced peroxisome proliferator-activated receptors transcriptional activity with EC 50 values of 18.26−31.45 μM. Conclusion: The anthraquinone derivatives were the active components from the roots of P. multiflorum as an inhibitor on inflammation-related factors in human hepatoma cells. Therefore, we suggest that the roots of P. multiflorum can be used to treat natural inflammatory diseases.
  2 1,301 19
Identification and quantification of the major constituents in Egyptian carob extract by liquid chromatography–electrospray ionization-tandem mass spectrometry
Asmaa Ibrahim Owis, El-Motaz Bellah El-Naggar
January 2016, 12(45):1-6
DOI:10.4103/0973-1296.176108  PMID:27041851
Background: Carob - Ceratonia siliqua L., commonly known as St John's-bread or locust bean, family Fabaceae - is one of the most useful native Mediterranean trees. There is no data about the chromatography methods performed by high performance liquid chromatography (HPLC) for determining polyphenols in Egyptian carob pods. Objective: To establish a sensitive and specific liquid chromatography–electrospray ionization (ESI)-tandem mass spectrometry (MSn) methodology for the identification of the major constituents in Egyptian carob extract. Materials and Methods: HPLC with diode array detector and ESI-mass spectrometry (MS) was developed for the identification and quantification of phenolic acids, flavonoid glycosides, and aglycones in the methanolic extract of Egyptian C. siliqua. The MS and MSn data together with HPLC retention time of phenolic components allowed structural characterization of these compounds. Peak integration of ions in the MS scans had been used in the quantification technique. Results: A total of 36 compounds were tentatively identified. Twenty-six compounds were identified in the negative mode corresponding to 85.4% of plant dry weight, while ten compounds were identified in the positive mode representing 16.1% of plant dry weight, with the prevalence of flavonoids (75.4% of plant dry weight) predominantly represented by two methylapigenin-O-pentoside isomers (20.9 and 13.7% of plant dry weight). Conclusion: The identification of various compounds present in carob pods opens a new door to an increased understanding of the different health benefits brought about by the consumption of carob and its products.
  1 2,237 21
Steroidal saponin diosgenin from Dioscorea bulbifera protects cardiac cells from hypoxia-reoxygenation injury through modulation of pro-survival and pro-death molecules
Karuppiah Shanmugasundarapandian Jayachandran, A Hannah Rachel Vasanthi, Narasimman Gurusamy
January 2016, 12(45):14-20
DOI:10.4103/0973-1296.176114  PMID:27041852
Background: Diosgenin, a steroidal saponin from plants, exhibits many biological potentials. Herein, the cardioprotective role of diosgenin is studied. Materials and Methods: The effect of diosgenin, isolated from Dioscorea bulbifera, was studied on hypoxia-reoxygenation (HR) in H9c2 cardiomyoblast cells. The amount of diosgenin in the plant extract was analyzed by high-performance thin layer chromatography using a solvent system comprising of chloroform:methanol:acetic acid:formic acid (13:4.5:1.5:1). Cardioprotection was checked by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Further, the release of lactate dehydrogenase, an enzyme released during cell death was checked. The proteins responsible for cell death (Bax) and cell survival (Bcl-2, hemeoxygenase-1 and Akt) were analyzed using Western blot to check the cardioprotective role of diosgenin. Conclusion:Supplementation of diosgenin mitigates HR injury, thereby exhibiting cardioprotective potential.
  1 2,002 19
Reverse phase-ultra flow liquid chromatography-diode array detector quantification of anticancerous and antidiabetic drug mangiferin from 11 species of Swertia from India
Parthraj R Kshirsagar, Nikhil B Gaikwad, Subhasis Panda, Harsha V Hegde, Sandeep R Pai
January 2016, 12(45):32-36
DOI:10.4103/0973-1296.176105  PMID:27041855
Background: Genus Swertia is valued for its great medicinal potential, mainly Swertia chirayita (Roxb. ex Fleming) H. Karst. is used in traditional medicine for a wide range of diseases. Mangiferin one of xanthoids is referred with enormous pharmacological potentials. Objective: The aim of the study was to quantify and compare the anticancerous and antidiabetic drug mangiferin from 11 Swertia species from India. The study also evaluates hierarchical relationships between the species based on mangiferin content using multivariate analysis. Materials and Methods: The reverse phase-ultra flow liquid chromatography-diode array detector analyses was performed and chromatographic separation was achieved on a Lichrospher 100, C18e (5 μm) column (250–4.6 mm). Mobile phase consisting of 0.2% triethylamine (pH-4 with O-phosphoric acid) and acetonitrile (85:15) was used for separation with injection volume 20 μL and detection wave length at 257 nm. Results: Results indicated that concentration of mangiferin has been found to vary largely between Swertia species collected from different regions. Content of mangiferin was found to be highest in Swertia minor compared to other Swertia species studied herein from the Western Ghats and Himalayan region also. The same was also evident in the multivariate analysis, wherein S. chirayita, S. minor and Swertia paniculata made a separate clade. Conclusion: Conclusively, the work herein provides insights of mangiferin content from 11 Swertia species of India and also presents their hierarchical relationships. To best of the knowledge this is the first report of higher content of mangiferin from any Swertia species.
  1 1,419 20
Endoplasmic reticulum stress-induced apoptotic pathway and mitochondrial dysregulation in HeLa cells treated with dichloromethane extract of Dillenia suffruticosa
Wan Abd Ghani Wan Nor Hafiza, Latifah Saiful Yazan, Yin Sim Tor, Jhi Biau Foo, Nurdin Armania, Heshu Sulaiman Rahman
January 2016, 12(45):86-95
DOI:10.4103/0973-1296.176107  PMID:27041866
Ethyl acetate and dichloromethane extract of Dillenia suffruticosa (EADS and DCMDS, respectively) can be a potential anticancer agent. The effects of EADS and DCMDS on the growth of HeLa cervical cancer cells and the expression of apoptotic-related proteins had been investigated in vitro. Cytotoxicity of the extracts toward the cells was determined by 5-diphenyltetrazolium bromide assay, the effects on cell cycle progression and the mode of cell death were analyzed by flow cytometry technique, while the effects on apoptotic-related genes and proteins were evaluated by quantitative real-time polymerase chain reaction, and Western blot and enzyme-linked immunosorbent assay, respectively. Treatment with DCMDS inhibited (P < 0.05) proliferation and induced apoptosis in HeLa cells. The expression of cyclin B1 was downregulated that led to G2/M arrest in the cells after treatment with DCMDA. In summary, DCMDS induced apoptosis in HeLa cells via endoplasmic reticulum stress-induced apoptotic pathway and dysregulation of mitochondria. The data suggest the potential application of DCMDS in the treatment of cervical cancer.
  1 1,504 26
Biological activities of Croton palanostigma Klotzsch
Eduardo Ferreira Mota, Diele Magno Rosario, Andreza Socorro Silva Veiga, Davi Do Socorro Barros Brasil, Fernando Tobias Silveira, Maria Fâni Dolabela
January 2016, 12(45):96-101
DOI:10.4103/0973-1296.176109  PMID:27041867
Background: Different species of Croton are used in traditional Amazonian medicine. Among the popular uses are treatment of bacterial diseases, poorly healing wounds and fevers. Objective: This study evaluated the antileishmanial, antiplasmodial and antimicrobial activities of the extracts and diterpenes of Croton palanostigma Klotzsch (Euphorbiaceae). Materials and Methods: Leaves and bark were extracted with dichloromethane and methanol. The bark dichloromethane extract (BDE) was chromatographed on a column, obtaining cordatin and aparisthman. The extracts and diterpenes were assayed thought agar disk diffusion method and their bactericidal or fungicidal effects were evaluated by minimum bactericidal or fungicidal concentration. The antiplasmodial activity was evaluated after 24 and 72 h of exposition. The antileishmanial activity was performed on promastigotes forms of Leishmania amazonensis. Results: The bark methanol extract (BME) and cordatin were not active against any microbial strains tested; BDE and leaves methanol extract (LME) were positive for Pseudomonas aeruginosa and aparisthman was positive for Candida albicans. In the determination of the minimum bactericidal concentration, neither of them were active in the highest concentration tested. The extracts and diterpenes were inactive in Plasmodium falciparum, except the LME in 72 h. Any extract was shown to be active in promastigote forms of L. amazonensis. Conclusion: These results indicate that the BDE and LME did not inhibit the bacterial growth, then they probably had bacteriostatic effect. LME presented activity in P. falciparum.
  - 1,002 21
Bioactivities of ethanolic extract and its fractions of Cistus laurifolius L. (Cistaceae) and Salvia wiedemannii Boiss. (Lamiaceae) species
Osman Ustun, Berrin-Ozcelik , Turhan Baykal
January 2016, 12(45):82-85
DOI:10.4103/0973-1296.176125  PMID:27041865
Background: Cistus laurifolius L. (Cistaceae) and Salvia wiedemannii Boiss. (Lamiaceae) have been used for treatment of some illnesses in Turkish folk medicine. In the present study, the ethanolic extract and its fractions obtained using re-extraction by hexane (Hx), chloroform (CHCl3), butanol, and remaining-water (r-H2O) of C. laurifolius were screened for their in vitro bioactivities. Materials and Methods: Activities were determined against both standard and the isolated strains of Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus faecalis, as well as yeasts such as Candida albicans and Candida parapsilosis by microdilution method. Also, antiviral activity of C. laurifolius and S. wiedemannii extracts were tested on herpes simplex virus-1 (HSV-1) and parainfluenza-3 (PI-3) using Madin-Darby bovine kidney and vero cell lines. Results: Tested extracts of C. laurifolius (minimum inhibitory concentration 32 μg/mL) exerted a strong antimicrobial activity against Gram-negative bacteria of E. coli, P. mirabilis, K. pneumoniae, and A. baumannii. Conclusion: The Hx extract of C. laurifolius (cytopathogenic effect of 32–8 μg/mL) had antiviral activity on PI-3. Also, the r-H2O, CHCl3, and ethanol extracts (16–<0.25 μg/mL) of S. wiedemannii had significant antiviral activity on HSV-1, same as control.
  - 1,324 20
Crispene A, B, C and D, four new clerodane type furanoid diterpenes from Tinospora crispa (L.)
Farhad Hossen, Rubaida Ahasan, Mohammad Rashedul Haque, Bilkis Begum, Choudhury Mahmood Hasan
January 2016, 12(45):37-41
DOI:10.4103/0973-1296.176116  PMID:27041856
Background: Tinospora crispa (L.) is used to alleviate the symptoms of diabetes mellitus in folk medicine. It is also used for hypertension and to treat malaria, remedy for diarrhea, and as vermifuge. Materials and Methods: Stems of T. crispa were collected, sun dried for several days followed by oven dried for 24 h at a considerably low temperature and then ground into coarse powder. The powdered stems were soaked in methanol at room temperature for 14 days with occasional shaking. The extract was collected by filtration, and the solvent was evaporated under reduced pressure in a rotary evaporator to obtain a solid residue which was then subjected to fractionation using the modified Kupchan partitioning method into n-hexane, CCl4, CHCl3and aqueous soluble fractions. The n-hexane soluble fraction was chromatographed over sephadex (LH-20) and the column was eluted with n-hexane: CH2Cl2:MeOH (2:5:1) followed by CH2Cl2:MeOH (9:1) and MeOH (100%) in order to increase the polarities. The column fractions were then concentrated and subjected to thin layer chromatography screening and the fractions with a satisfactory resolution of compounds were rechromatographed over silica gel to isolate the pure compounds. Results: Four new furanoid diterpenes of clerodane types, Crispene A, B, C, and D (1–4), including one known furanoid diterpene glucoside, borapetoside E (5), were isolated from the stems of T. crispa. The structures of these compounds were elucidated by means of extensive spectroscopic analysis and by comparison of their spectral data with closely related compounds. Conclusion: We have reported four new furanoid diterpenes of clerodane types, including one known furanoid diterpene glucoside. This is the first report of any clerodane diterpene having olefinic bond between C-6 and C-7.
  - 1,722 20
Essential oil from the resin of Protium heptaphyllum: Chemical composition, cytotoxicity, antimicrobial activity, and antimutagenicity
Ewelyne Miranda de Lima, Didley Sâmia Paiva Cazelli, Fernanda Endringer Pinto, Renata Alves Mazuco, Ieda Carneiro Kalil, Dominik Lenz, Rodrigo Scherer, Tadeu Uggere de Andrade, Denise Coutinho Endringer
January 2016, 12(45):42-46
DOI:10.4103/0973-1296.176113  PMID:27041857
Background: Protium heptaphyllum (Aubl.) March is popularly used as an analgesic and anti-inflammatory agent. Objective: This study aimed to evaluate the chemical composition of P. heptaphyllum essential oil, its cytotoxicity in a breast cancer cell line (MCF-7), antimicrobial activity, and its antimutagenicity in vivo. Materials and Methods: The chemical composition of the essential oil collected in three 3 years was determined by gas chromatography-mass spectrometry. The cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Annexin V conjugated with fluorescein isothiocyanate, caspase-3, and tumor necrosis factor-alpha (TNF-α) assays were performed to evaluate apoptosis and inflammatory events. The antimutagenic activity at doses of 25, 50, and 100 mg/kg was determined using a micronucleus test in murine bone marrow. Results: The essential oil showed a predominance of monoterpene compounds, being the terpinolene, p-cymene-8-ol, and p-cymene, present in the essential oil extracted in the 3 years. The essential oil showed a protection against cyclophosphamide-induced genotoxicity, and the cytotoxicity index polychromatic erythrocytes/normochromatic erythrocytes ratio in animals treated with oil at all doses (1.34 ± 0.33; 1.15 ± 0.1; 1.11 ± 0.13) did not differ from the negative control animal (1.31 ± 0.33), but from the cyclophosphamide group (0.61 ± 0.12). Cytotoxicity, at a concentration of 40.0 μg/mL, and antimicrobial activity were not observed for the essential oil (minimum inhibitory concentration ≥0.5 mg/mL). The essential oil did not change the levels of caspase-3 in the TNF-α level. Conclusion: The essential oil showed antimutagenic activity due to its chemical composition.
  - 2,058 22
New isorhamnetin derivatives from Salsola imbricata Forssk. leaves with distinct anti-inflammatory activity
Samir M Osman, Walaa A El Kashak, Michael Wink, Mohamed A El Raey
January 2016, 12(45):47-51
DOI:10.4103/0973-1296.176110  PMID:27041858
Background: Salsola imbricata Forssk. is a shrub widely growing in Egypt, used as a camel food, traditionally, used as anti-inflammatory agent. Literature survey showed no report about the anti-inflammatory activity of S. imbricata. Aim of the Study: This work was designed to study the phenolic constituents and to provide evidence for the traditional use of S. imbricata as an anti-inflammatory agent. Materials and Methods: The in vitro anti-inflammatory activity of the total aqueous methanol extract and some isolated compounds were investigated in RAW 264.7 macrophage cells using nitric oxide assay. All chemical structures were identified on the basis of electrospray ionization-mass spectrometry, one- and two-dimension nuclear magnetic resonance. Results: Nine phenolic compounds, among them two new natural products; isorhamnetin-3-O-β-D-glucuronyl (1'''→4'') glucuronide (1) and its dimethyl ester; isorhamnetin-3-O-β-D-di glucuronate dimethyl ester (2), two isorhamnetin glycosides: Isorhamnetin-3-O-β-D-galactopyranoside (3), isorhamnetin-3-O-β-D-glucopyranoside (4), and isorhamnetin (5). In addition, an alkaloidal phenolic; trans N-feruloyl tyramine (6), three phenolic acids: Isovanillic acid (7), ferulic acid (8), and p-hydroxy benzoic acid (9) were isolated from salsola imbricata leaves. All compounds were isolated and identified for the first time from this plant except compound (6). The extract and the tested compounds showed distintict anti-inflammatory activities with no toxicity on RAW 264.7 macrophage cells. Conclusion: The extract and the tested compounds showed distintict anti-inflammatory activities with no toxicity on RAW 264.7 macrophage cells.
  - 1,410 20
Cytotoxic properties and complete nuclear magnetic resonance assignment of isolated xanthones from the root of Garcinia cowa Roxb.
Fatma Sri Wahyuni, Khozirah Shaari, Johnson Stanslas, Nordin HJ Lajis, Hamidi Dachriyanus
January 2016, 12(45):52-56
DOI:10.4103/0973-1296.176115  PMID:27041859
Objective: To isolate compounds from the roots of Garcinia cowa and to evaluated their cytotoxic activity against breast (MCF-7), prostate (DU-145), and lung (H-460) cell lines. Materials and Methods: The ground air-dried root was sequentially macerated with hexane, dichloromethane (DCM), ethyl acetate (EtOAc), and methanol. The DCM soluble extract was fractionated by vacuum liquid chromatography, column chromatography, and radial chromatography over silica gel with hexane, EtOAc and methanol as eluent in progressively increasing polarity manner; to yield three compounds. Their structures were elucidated based on their spectroscopic data and their comparison with those of the literature. The cytotoxicity of isolated compounds was carried out against human cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. The extract was added at various concentrations (0.1, 1, 10 and 100 mg/ml). The level of cytotoxicity was determined by calculating the level of IC50 that was based on the percentage of the cell death following the 24 h incubation with the extract. Results: Phytochemical study on the roots of G. cowa yielded rubraxanthone (3), cowanine (4) and 1,5-dihydroxyxanthone (5). Compound 4 with an IC50 value of 4.1 ± 1.0 μM, 5.4 ± 2.3 μM and 11.3 ± 10.0 μM against MCF-7, H-460, and DU-145, respectively while compound 3 was found to be in active. Conclusion: The results indicate that G. cowa roots could be important sources of natural cytotoxic compounds.
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New synthetic pyrazine carboxamide derivatives as potential elicitors in production of secondary metabolite in In vitro cultures
Lenka Tůmová, Jiří Tůma, Martin Doležal, Zuzana Dučaiová, Jan Kubeš
January 2016, 12(45):57-62
DOI:10.4103/0973-1296.176117  PMID:27041860
Background: Silymarin, an active polyphenolic fraction of Silybum marianum, and high flavonoid content of Fagopyrum possess various interesting biological activities. The substituted pyrazine-2-carboxamides were previously used as effective elicitors of studied secondary metabolites. Objective: To study the effect of new synthetic pyrazine carboxamide derivatives, N-(4-chlorobenzyl)-5-tert-butylpyrazine-2-carboxamide (1) and 3-(3-((trifluoromethyl) benzyl) amino) pyrazine-2-carboxamide (2), on flavonolignan and flavonoid production in S. marianum and Fagopyrumes culentum in vitro cultures. Materials and Methods: Callus and suspension cultures were cultured on MS medium containing α-naphtaleneacetic acid or 2,4-D. Three elicitor concentrations for different exposure times were tested. Dried and powdered samples of callus and suspension cultures were extracted with methanol and analyzed by DAD-HPLC. Results: Compound 1 showed as a good elicitor of taxifolin production. The effect on silymarin complex was less visible with a maximum between 24 and 48 h after 3.292 ×10−4 mol/L concentration. The detailed analysis showed that silychristin was the most abundant. Compound 2 was effective in rutin production only in callus culture with maximum 24 h and 168 h after application of 3.3756 ×10−3 mol/L concentration and 48 and 72 h after 3.3756 ×10−4 mol/L concentration. Conclusion: From the results of the performed experiments, it can be concluded that compound 1 shows to be suitable elicitor for enhanced production of taxifolin and silychristin in S. marianum, mainly when 3.292 ×10−4 mol/L concentration was used, and compound 2 is suitable for increase rutin production in callus cultures and less appropriate for suspension cultures of F. esculentum.
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Alteration in memory and electroencephalogram waves with sub-acute noise stress in albino rats and safeguarded by Scoparia dulcis
Sundareswaran Loganathan, Sheeladevi Rathinasamy
January 2016, 12(45):7-13
DOI:10.4103/0973-1296.176119  PMID:27041862
Background: Noise stress has different effects on memory and novelty and the link between them with an electroencephalogram (EEG) has not yet been reported. Objective: To find the effect of sub-acute noise stress on the memory and novelty along with EEG and neurotransmitter changes. Materials and Methods: Eight-arm maze (EAM) and Y-maze to analyze the memory and novelty by novel object test. Four groups of rats were used: Control, control treated with Scoparia dulcis extract, noise exposed, and noise exposed which received Scoparia extract. Results: The results showed no marked difference observed between control and control treated with Scoparia extract on EAM, Y-maze, novel object test, and EEG in both prefrontal and occipital region, however, noise stress exposed rats showed significant increase in the reference memory and working memory error in EAM and latency delay, triad errors in Y-maze, and prefrontal and occipital EEG frequency rate with the corresponding increase in plasma corticosterone and epinephrine, and significant reduction in the novelty test, and significant reduction in the novelty test, amplitude of prefrontal, occipital EEG, and acetylcholine. Conclusion: These noise stress induced changes in EAM, Y-maze, novel object test, and neurotransmitters were significantly prevented when treated with Scoparia extract and these changes may be due to the normalizing action of Scoparia extract on the brain, which altered due to noise stress.
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Hybrid analysis (barcode-high resolution melting) for authentication of Thai herbal products, Andrographis paniculata (Burm.f.) Wall.ex Nees
Maslin Osathanunkul, Chatmongkon Suwannapoom, Nuttaluck Khamyong, Danupol Pintakum, Santisuk Na Lamphun, Kanokporn Triwitayakorn, Kitisak Osathanunkul, Panagiotis Madesis
January 2016, 12(45):71-75
DOI:10.4103/0973-1296.176112  PMID:27041863
Background: Andrographis paniculata Nees is a medicinal plant with multiple pharmacological properties. It has been used over many centuries as a household remedy. A. paniculata products sold on the markets are in processed forms so it is difficult to authenticate. Therefore buying the herbal products poses a high-risk of acquiring counterfeited, substituted and/or adulterated products. Due to these issues, a reliable method to authenticate products is needed. Materials and Methods: High resolution melting analysis coupled with DNA barcoding (Bar-HRM) was applied to detect adulteration in commercial herbal products. The rbc L barcode was selected to use in primers design for HRM analysis to produce standard melting profile of A. paniculata species. DNA of the tested commercial products was isolated and their melting profiles were then generated and compared with the standard A. paniculata. Results: The melting profiles of the rbc L amplicons of the three closely related herbal species (A. paniculata, Acanthus ebracteatus and Rhinacanthus nasutus) are clearly separated so that they can be distinguished by the developed method. The method was then used to authenticate commercial herbal products. HRM curves of all 10 samples tested are similar to A. paniculata which indicated that all tested products were contained the correct species as labeled. Conclusion: The method described in this study has been proved to be useful in aiding identification and/or authenticating A. paniculata. This Bar-HRM analysis has allowed us easily to determine the A. paniculata species in herbal products on the markets even they are in processed forms.
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Optimization of process parameters and kinetic model of enzymatic extraction of polyphenols from Lonicerae flos
Fansheng Kong, Shujuan Yu, Yongguang Bi, Xiaojun Huang, Mengqian Huang
January-March 2016, 12(45):70-74
DOI:10.4103/0973-1296.176055  PMID:27018039
Objective: To optimize and verify the cellulase extraction of polyphenols from honeysuckle and provide a reference for enzymatic extracting polyphenols from honeysuckle. Materials and Methods: The uniform design was used According to Fick's first law and kinetic model, fitting analysis of the dynamic process of enzymatic extracting polyphenols was conducted. Results: The optimum enzymatic extraction parameters for polyphenols from honeysuckle are found to be 80% (v/v) of alcohol, 35:1 (mL/g) of liquid-solid ratio, 80°C of extraction temperature, 8.5 of pH, 6.0 mg of enzyme levels, and 130 min of extraction time. Under the optimal conditions, the extraction rate of polyphenols was 3.03%. The kinetic experiments indicated kinetic equation C / CC0 had a good linear relationship with t even under the conditions of different levels of enzyme and temperature, which means fitting curve tallies well with the experimental values. Conclusion: The results of quantification showed that the results provide a reference for enzymatic extracting polyphenols from honeysuckle.
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Molecular docking analysis of selected Clinacanthus nutans constituents as xanthine oxidase, nitric oxide synthase, human neutrophil elastase, matrix metalloproteinase 2, matrix metalloproteinase 9 and squalene synthase inhibitors
Radhakrishnan Narayanaswamy, Azizul Isha, Lam Kok Wai, Intan Safinar Ismail
January 2016, 12(45):21-26
DOI:10.4103/0973-1296.176111  PMID:27041853
Background: Clinacanthus nutans (Burm. f.) Lindau has gained popularity among Malaysians as a traditional plant for anti-inflammatory activity. Objective: This prompted us to carry out the present study on a selected 11 constituents of C. nutans which are clinacoside A–C, cycloclinacoside A1, shaftoside, vitexin, orientin, isovitexin, isoorientin, lupeol and β-sitosterol. Materials and Methods: Selected 11 constituents of C. nutans were evaluated on the docking behavior of xanthine oxidase (XO), nitric oxide synthase (NOS), human neutrophil elastase (HNE), matrix metalloproteinase (MMP 2 and 9), and squalene synthase (SQS) using Discovery Studio Version 3.1. Also, molecular physicochemical, bioactivity, absorption, distribution, metabolism, excretion, and toxicity (ADMET), and toxicity prediction by computer assisted technology analyzes were also carried out. Results: The molecular physicochemical analysis revealed that four ligands, namely clinacoside A–C and cycloclinacoside A1 showed nil violations and complied with Lipinski's rule of five. As for the analysis of bioactivity, all the 11 selected constituents of C. nutans exhibited active score (>0) toward enzyme inhibitors descriptor. ADMET analysis showed that the ligands except orientin and isoorientin were predicted to have Cytochrome P4502D6 inhibition effect. Docking studies and binding free energy calculations revealed that clinacoside B exhibited the least binding energy for the target enzymes except for XO and SQS. Isovitexin and isoorientin showed the potentials in the docking and binding with all of the six targeted enzymes, whereas vitexin and orientin docked and bound with only NOS and HNE. Conclusion: This present study has paved a new insight in understanding these 11 C. nutans ligands as potential inhibitors against XO, NOS, HNE, MMP 2, MMP 9, and SQS.
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Potential antidepressant constituents of Nigella sativa seeds
Ehab S Elkhayat, Mohammad S Alorainy, Ibrahim M El-Ashmawy, Shawkat Fat'hi
January 2016, 12(45):27-31
DOI:10.4103/0973-1296.176118  PMID:27041854
Background: Nigella sativa Linn. is well known seed in the Middle East, Asia, and the Far East as a natural remedy for many ailments and as a flavoring agent proclaimed medicinal usage dating back to the ancient Egyptians, Greeks, and Romans. An authentic saying of the Prophet Muhammad (Peace Be Upon Him) about black seed is also quoted in Al-Bukhari. Objective: This study was carried out to evaluate the antidepressant effect and isolate the potential antidepressant constituents of the polar extract of N. sativa seeds. Materials and Methods: The antidepressant effect was evaluated through the immobility duration in tail suspension and forced swim tests (FSTs). Albino mice were orally treated with N. sativa polar extract and its RP-18 column chromatography fractions (50 and 100 mg/kg,). Results: The polar extract and two of its sub-fractions were significantly able to decrease the immobility time of mice when subjected to both tail suspension and FSTs, the effects are comparable to standard drug (Sertraline, 5 mg/kg). However, these treatments did not affect the number of crossings and rearing in the open field test. Phytochemical investigation of the two active fractions led to the isolation of quercetin-3-O-α-L-rhamnopyranoside 1, quercetin-7-O-β-D-gluco- pyranoside 2, tauroside E 3, and sapindoside B as the potential antidepressant constituents.
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Determination and pharmacokinetic comparisons of atractylodin after oral administration of crude and processed Atractylodis rhizoma
Chang Xiao-Wen, Xu Chen-Xi, Liu Yu-Qiang, Qian Cai
January-March 2016, 12(45):80-83
DOI:10.4103/0973-1296.176062  PMID:27019565
Background: In traditional Chinese medicine, Atractylodis rhizoma is the dried rhizome of Atractylodes lancea (Thunb.) DC. or Atractylodes chinensis (DC.) Koidz. After being processed, the dryness of A. rhizoma decreased, and the function of tonifying spleen increased. Therefore, the processed A. rhizoma is the best choice of clinical application. As the main active components, polyethylene alkynes exhibits various desirable pharmacological effects including anti-inflammatory, anti-bacterial and anti-arrhythmia activity. However, there is no report on the pharmacokinetic comparisons of atractylodin, one of polyethylene alkynes, in bio-samples after oral administration of crude and processed A. rhizoma until now. The in vivo study of active components of A. rhizoma would be necessary and helpful for clinical application and clarification of processing principle. Objective: To compare the pharmacokinetic parameters of atractylodin after oral administration of crude and processed A. rhizoma, and clarify the processing principle of A. rhizoma. Materials and Methods: Plasma concentrations of atractylodin in rats were determined by reversed-phase high-performance liquid chromatogram and the main pharmacokinetic parameters were estimated with Drug and Statistics 2.0 Software Package (Chinese Pharmacological Society, Shanghai, China). Results: The AUC 0−t, AUC 0→∞ , T max, and C max of processed A. rhizoma were increased significantly (P < 0.05) compared with that in crude A. rhizoma after using Student's t-test. Conclusions: Processing A. rhizoma with wheat bran by stir-frying can promote and accelerate the absorption of atractylodin.
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H2O2 improves quality of Radix scutellariae through anti-oxidant effect
Song Qi, Cao Wu-lin, Jiang Hua, Zhang Ai-hua, Meng Xiang-cai
January-March 2016, 12(45):84-90
DOI:10.4103/0973-1296.176063  PMID:27019566
Introduction: The correlation between the quality and geographical origin of herbal medicine was traced back to Tang Dynasty in China, more than 1200 years, and the effects of ecological environments on the secondary metabolites such as flavonoids have been confirmed. However, little is known about how the adversity impacts on the quality. Reactive oxygen species (ROS) may be medium between the ecological environment and the secondary metabolism. Materials and Methods: The fresh roots of Scutellaria baicalensis Georgi were treated with 0.002 μmol/L, 0.2 μmol/L, and 20 μmol/L H 2 O 2, respectively. A stress model was established to elucidate the change of secondary metabolism, anti-oxidant enzyme system, and enzymes relating to flavonoids. Results: The activities of superoxide dismutase, catalase and peroxidase decreased. Too much H 2 O 2, firstly, boosted transformation of flavonoids glycoside into aglucon with the most remarkable activities through UDP-glucuronate baicalein 7-O-glucuronosyltransferase (UBGAT), and β-glucuronidase (GUS), then regulated the gene expression of phenylalanine ammonialyase, GUS, and UBGAT, and increased the contents of flavones, motivated the flavonoid glycoside converting into aglucon. With this action, the flavones displaced the anti-oxidant enzymes. The higher the dosage, the more baicalein and wogonin increased, the later they took action. Conclusion: The plant secondary metabolites to keep ROS constant are identical to the effective materials in clinic. They are closely linked. H 2 O 2 can improve flavones, especially the aglucon, and further increased the quality of herbal medicine, which possesses very important value in medical practice.
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Optimization of ultrasound-assisted extraction of phenolic compounds from Myrcia amazonica DC. (Myrtaceae) leaves
Mariana Cristina de Morais Rodrigues, Leonardo Luiz Borges, Frederico Severino Martins, Rosa Helena V Mourão, Edemilson Cardoso da Conceição
January-March 2016, 12(45):9-12
DOI:10.4103/0973-1296.175997  PMID:27019555
Background: Myrcia amazonica. DC is a species predominantly found in northern Brazil, and belongs to the Myrtaceae family, which possess various species used in folk medicine to treat gastrointestinal disorders, infectious diseases, and hemorrhagic conditions and are known for their essential oil contents. Materials and Methods: This study aimed applied the Box-Behnken design combined with response surface methodology to optimize ultrasound-assisted extraction of total polyphenols, total tannins (TT), and total flavonoids (TF) from M. amazonica DC. Results: The results indicated that the best conditions to obtain highest yields of TT were in lower levels of alcohol degree (65%), time (15 min), and also solid: Liquid ratio (solid to liquid ratio; 20 mg: 5 mL). The TF could be extracted with high amounts with higher extraction times (45 min), lower values of solid: Liquid ratio (20 mg: mL), and intermediate alcohol degree level. Conclusion: The exploitation of the natural plant resources present very important impact for the economic development, and also the valorization of great Brazilian biodiversity. The knowledge obtained from this work should be useful to further exploit and apply this raw material.
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7-hydroxycalamenene effects on secreted aspartic proteases activity and biofilm formation of Candida spp.
Mariana M. B. Azevedo, Catia A Almeida, Francisco C. M. Chaves, Igor A Rodrigues, Humberto R Bizzo, Celuta S Alviano, Daniela S Alviano
January-March 2016, 12(45):36-40
DOI:10.4103/0973-1296.176022  PMID:27019560
Background: The 7-hydroxycalamenenene-rich essential oil (EO) obtained from the leaves of Croton cajucara (red morphotype) have been described as active against bacteria, protozoa, and fungi species. In this work, we aimed to evaluate the effectiveness of 7-hydroxycalamenenene against Candida albicans and nonalbicans species. Materials and Methods: C. cajucara EO was obtained by hydrodistillation and its major compound, 7-hydroxycalamenene, was purified using preparative column chromatography. The anti-candidal activity was investigated by minimum inhibitory concentration (MIC) and secreted aspartic proteases (SAP) and biofilm inhibition assays. Results: 7-hydroxycalamenene (98% purity) displayed anti-candidal activity against all Candida species tested. Higher activity was observed against Candida dubliniensis, Candida parapsilosis and Candida albicans, showing MIC values ranging from 39.06 μg/ml to 78.12 μg/ml. The purified 7-hydroxycalamenene was able to inhibit 58% of C. albicans ATCC 36801 SAP activity at MIC concentration (pH 7.0). However, 7-hydroxycalamenene demonstrated poor inhibitory activity on C. albicans ATCC 10231 biofilm formation even at the highest concentration tested (2500 μg/ml). Conclusion: The bioactive potential of 7-hydroxycalamenene against planktonic Candida spp. further supports its use for the development of antimicrobials with anti-candidal activity.
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An efficient high-performance liquid chromatography combined with electrospray ionization tandem mass spectrometry method to elaborate the changes of components between the raw and processed radix Aconitum kusnezoffii
Beibei Wang, Jiaojiao Ji, Shuang Zhao, Jie Dong, Peng Tan, Shengsang Na, Yonggang Liu
January-March 2016, 12(45):4-8
DOI:10.4103/0973-1296.175989  PMID:27019554
Background: Crude radix Aconitum kusnezoffii (RAK) has great toxicity. Traditional Chinese medicine practice proved that processing may decrease its toxicity. In our previous study, we had established a new method of RAK processing (Paozhi). However, the mechanism is yet not perfect. Objective: To explore the related mechanism of processing through comparing the chemical contents. Materials and Methods: A new processing method of RAK named stoving (Hong Zhi) was used. In particular, RAK was stored at 110°C for 8 h, and then high performance liquid chromatography combined with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS n ) was developed for the detection of the alkaloids of the crude and processed RAK decoction pieces. Results: Thirty components of the crude RAK were discovered, among which, 23 alkaloids were identified. Meanwhile, 23 ingredients were detected in the processed RAK decoction pieces, among which, 20 alkaloids were determined yet. By comparison, eight alkaloids were found in both crude and processed RAK decoction pieces, 15 alkaloids were not found in the crude RAK, however, 10 new constituents yield after processing, which are 10-OH-hypaconine, 10-OH-mesaconine, isomer of bullatine A, 14-benzoyl-10-OH-mesaconine, 14-benzoyl-10-OH-aconine, 14-benzoyl-10-OH-hypaconine, dehydrated aconitine, 14-benzoylaconine, chuanfumine, dehydrated mesaconitine. Conclusion: The present study showed that significant change of alkaloids was detected in RAK before and after processing. Among them, the highly toxic diester alkaloids decreased and the less toxic monoester alkaloids increased. Moreover, the concentration changes significantly. HPLC-ESI-MS n are Efficient to elaborate the mechanism of reduction of toxicity and enhancement efficacy after processing.
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Influence of chitosan nanoparticles as the absorption enhancers on salvianolic acid B In vitro and In vivo evaluation
Xin Jin, Shi-bing Zhang, Shi-meng Li, Ke Liang, Zeng-yong Jia
January-March 2016, 12(45):57-63
DOI:10.4103/0973-1296.176047  PMID:27019562
Background: Salvianolic acid B (SalB) represents the most abundant and bio-active phenolic constituent among the water-soluble compounds of Salvia miltiorrhiza. But the therapeutic potential of SalB has been significantly restricted by its poor absorption. Methods: In this study, chitosans (CS) and CS nanoparticles (NPs) with different molecular weights (MWs), which have influence on the absorption of SalB, was also investigated. Results: As a preliminary study, water-soluble CS with various MWs (3, 30, 50, and 100 kDa) was chosen. We investigated the MW-dependent Caco-2 cell layer transport phenomena in vitro of CS and NPs at concentrations (4 μg/ml, w/v). SalB, in presence CS or NPs has no significant toxic effect on Caco-2 cell. As the MW increases, the absorption enhancing effect of CS increases. However, as the MW decreases, the absorption enhancing effect of NPs increases. The AUC 0-∞ of the SalB-100 kDa CS was 4.25 times greater than that of free SalB. And the AUC 0-∞ of the SalB-3 kDa NPs was 16.03 times greater than that of free SalB. Conclusion: CS and NPs with different MWs as the absorption enhancers can promote the absorption of SalB. And the effect on NPs is better than CS.
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Luffa echinata Roxb. induced apoptosis in human colon cancer cell (SW-480) in the caspase-dependent manner and through a mitochondrial apoptosis pathway
Li-Hua Shang, Yan Yu, De-Hai Che, Bo Pan, Shi Jin, Xiao-Long Zou
January-March 2016, 12(45):25-30
DOI:10.4103/0973-1296.176017  PMID:27019558
Background: Luffa echinata Roxb. (LER) (Cucurbitaceae) showed tremendous medicinal importance and are being used for the treatment of different ailments. Objective: In this study, the antiproliferative properties and cell death mechanism induced by the extract of the fruits of LER were investigated. Materials and Methods: MTT and LDH assay were used to test the antiproliferative and cytotoxicity of LER extract, respectively. The intracellular ROS were measured by a fluorometric assay. The expression of several apoptotic-related proteins in SW-480 cells treated by LER was evaluated by Western blot analysis. Results: The methanolic extract of LER fruits inhibited the proliferation of human colon cancer cells (SW-480) in both dose- and time-dependent manners. The LER-treated cells showed obvious characteristics of cell apoptosis, including cell shrinkage, destruction of the monolayer, and condensed chromatin. In addition, treatments of various concentrations of LER extracts caused the release of lactate dehydrogenase as a dose-dependent manner via stimulation of the intracellular metabolic system. LER induced apoptosis, DNA fragmentation, and cellular ROS accumulation in SW-480 cells. Treatment of LER on SW-480 cells promoted the expression of caspases, Bax, Bad, and p53 proteins and decreased the levels of Bcl-2 and Bcl-XL. Conclusions: These results indicated that treatment with LER-induced cell death in mitochondrial apoptosis pathway by regulating pro-apoptotic proteins via the up regulation of the p53 protein. These findings highlight the potentials of LER in the treatment of human colon cancer.
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A new biphenyl neolignan from leaves of Patrinia villosa (Thunb.) Juss.
Yan Xin-Jia, Liu Wei, Zhao Ying, Chen Ning, Xu Ying, Wu Jian, Wang Tan, Li Yue, Xiang Zheng
January-March 2016, 12(45):1-3
DOI:10.4103/0973-1296.175988  PMID:27019553
Results: One new stereoisomer of biphenylneolignan with four known compounds was isolated from the leaves of Patrinia villosa Juss. Methods: The structure of the new compound was elucidated as 2,6,2',6'-tetramethoxy-4,4'-bis (1,2-trans-2,3-epoxy-1-hydroxypropyl) biphenyl (1) on the basis of spectroscopic analysis and comparison with literature data. The four known compounds were identified as 2,6,2',6'-tetramethoxy-4,4'-bis(1,2-cis-2,3-epoxy-1-hydroxypropyl)biphenyl (2), 1H-indole-3-carbaldehyde (3), luteolin (4) and quercetin(5) by comparison of their spectral data with the reported data, respectively. Conclusions: Compound 1 is a new biphenylneolignan, compound 2 and 3 were isolated for the first time from the plant.
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Determination of ruscogenin in ophiopogonis radix by high-performance liquid chromatography-evaporative light scattering detector coupled with hierarchical clustering analysis
Chun-Hua Liu, Ming Li, Ya-Qian Feng, Yuan-Jia Hu, Bo-Yang Yu, Jin Qi
January-March 2016, 12(45):13-20
DOI:10.4103/0973-1296.176008  PMID:27019556
Background: Ophiopogonis Radix is a famous traditional Chinese medicine. It is necessary to establish a suitable quality control methods of Ophiopogonis Radix. Objective: To investigate the quality control methods of Ophiopogonis Radix by high-performance liquid chromatography (HPLC) coupled with evaporative light scattering detector (ELSD). Materials and Methods: A rapid and simple method, HPLC coupled with ELSD, was applied to determinate ruscogenin in 35 batches of Ophiopogenis Radix samples. Orthogonal tests and single factor explorations were used to optimize the extraction condition of ruscogenin. The content of ruscogenin in different origin was further analyzed by hierarchical clustering analysis (HCA). Results: The ruscogenin was successfully determined by HPLC-ELSD with a two-phase solvent system composed of methanol-water (88:12) at a flow rate 1.0 ml/min, column temperature maintained at 25°C, detector draft tube temperature at 42.2°C, nebulizer gas flow rate at 1.4 L/min, and the gain at 8. The result showed the good linearity of ruscogenin in the range of 40.20-804.00 μg/ml (R2 = 0.9996). Average of recovery was 101.3% (relative standard deviation = 1.59%). A significant difference of ruscogenin content was shown among 35 batches of Ophiopogenis Radix from different origin, varied from 0.0035% to 0.0240%. HCA based on the content of ruscogenin indicated that Ophiopogonis Radix in different origin was mainly divided into two clusters. Conclusion: This simple, rapid, low-cost, and reliable HPLC-ELSD method could be suitable for measurement of ruscogenin content rations and quality control of Ophiopogonis Radix.
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