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Table of Contents
May 2015
Volume 11 | Issue 42 (Supplement)
Page Nos. 1-230
Online since Wednesday, May 27, 2015
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ORIGINAL ARTICLES
Cytotoxic glucosphingolipid from
Celtis Africana
p. 1
Shagufta Perveen, Areej Mohammad Al-Taweel, Ghada Ahmed Fawzy, Azza Muhammed El-Shafae, Afsar Khan, Peter Proksch
DOI
:10.4103/0973-1296.157662
PMID
:26109753
Background:
Literature survey proved the use of the powdered sun-dried bark and roots of
Celtis africana
for the treatment of cancer in South Africa.
Objective:
The aim of this study was to do further isolation work on the ethyl acetate fraction and to investigate the cytotoxic activities of the various fractions and isolated compound.
Materials and Methods:
Cytotoxicity of petroleum ether, chloroform, ethyl acetate,
n
-butanol fractions and compound 1 were tested on mouse lymphoma cell line L5178Y using the microculture tetrazolium assay.
Results:
One new glucosphingolipid 1 was isolated from the aerial parts of
C. africana
. The structure of the new compound was determined by extensive analysis by one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy and mass spectrometry. The ethyl acetate fraction and compound 1 showed strong cytotoxic activity with an EC
50
value of 8.3 μg/mL and 7.8 μg/mL, respectively, compared with Kahalalide F positive control (6.3 μg/mL).
Conclusion:
This is the first report of the occurrence of a cytotoxic glucosphingolipid in family
Ulmaceae
.
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Cytotoxicity of some edible mushrooms extracts over liver hepatocellular carcinoma cells in conjunction with their antioxidant and antibacterial properties
p. 6
Gökhan Sadi, Bugrahan Emsen, Abdullah Kaya, Aytaç Kocabas, Seval Çinar, Deniz Irtem Kartal
DOI
:10.4103/0973-1296.157665
PMID
:26109775
Background:
Mushrooms have been valued for their nutritive content and as traditional medicines; several important medicinal properties of mushrooms have been recognized worldwide.
Objective:
The purpose of this study was to elucidate the cell growth inhibitory potential of four edible mushrooms;
Coprinus comatus
(O.F. Mull.) Pers. (Agaricaceae),
Tricholoma fracticum
(Britzelm.) Kreisel (Tricholomataceae),
Rhizopogon luteolus
Fr. and Nordholm (Rhizopogonaceae),
Lentinus tigrinus
(Bull.) Fr. (Polyporaceae) on hepatocellular carcinoma (HepG2) cells in conjunction with their antioxidant and antibacterial capacities.
Materials
and
Methods:
Five different extracts of edible mushrooms were obtained using water, methanol, acetone,
n
-hexane and chloroform as solvent systems for cytotoxic, antioxidant and antibacterial properties.
Results:
C. comatus
showed substantial
in vitro
cytotoxic activity against HepG2 cell lines with all extracts especially with chloroform 50% inhibition (IC
50
value of 0.086 mg/ml) and acetone (IC
50
value of 0.420 mg/ml). Chloroform extract of
C. comatus
had maximum amount of β-carotene (25.94 μg/mg), total phenolic content (76.32 μg/mg) and lycopene (12.00 μg/mg), and
n
-hexane extract of
L. tigrinus
had maximum amount of flavonoid (3.67 μg/mg). While chloroform extract of
C. comatus
showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) capturing activity (1.579 mg/ml), the best result for metal chelating activity was obtained from methanolic extract (0.842 mg/ml). Moreover, all tested mushrooms demonstrated antibacterial activity and
n
-hexane extract of
L. tigrinus
and acetone extracts of
T. fracticum
were the most active against tested microorganism.
Conclusion:
These results indicate that different extracts of investigated mushroom have considerable cytotoxic, antioxidant and antibacterial properties and may be utilized as a promising source of therapeutics.
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Pharmacophore modeling,
in silico
screening, molecular docking and molecular dynamics approaches for potential alpha-delta bungarotoxin-4 inhibitors discovery
p. 19
R Barani Kumar, M Xavier Suresh, B Shanmuga Priya
DOI
:10.4103/0973-1296.157670
PMID
:26109766
Background:
The alpha-delta bungartoxin-4 (α-δ-Bgt-4) is a potent neurotoxin produced by highly venomous snake species,
Bungarus caeruleus,
mainly targeting neuronal acetylcholine receptors (nAchRs) and producing adverse biological malfunctions leading to respiratory paralysis and mortality. Objective: In this study, we predicted the three-dimensional structure of α-δ-Bgt-4 using homology modeling and investigated the conformational changes and the key residues responsible for nAchRs inhibiting activity.
Materials and Methods:
From the selected plants, which are traditionally used for snake bites, the active compounds are taken and performed molecular interaction studies and also used for modern techniques like pharmacophore modeling and mapping and absorption, distribution, metabolism, elimination and toxicity analysis which may increase the possibility of success.
Results:
Moreover, 100's of drug-like compounds were retrieved and analyzed through computational virtual screening and allowed for pharmacokinetic profiling, molecular docking and dynamics simulation.
Conclusion:
Finally the top five drug-like compounds having competing level of inhibition toward α-δ-Bgt-4 toxin were suggested based on their interaction with α-δ-Bgt-4 toxin.
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Antimutagenic potential of harpagoside and
Harpagophytum procumbens
against 1-nitropyrene
p. 29
Luigi Manon, Baghdikian Béatrice, Orsičre Thierry, Pompili Jocelyne, Mabrouki Fathi, Ollivier Evelyne, Botta Alain
DOI
:10.4103/0973-1296.157675
PMID
:26109771
Background:
1-nitropyrene (1-NPy) is one of the most abundant nitro-polycyclic aromatic hydrocarbons particularly in diesel exhausts. It is a mutagenic and carcinogenic pollutant very widespread in the environment. So the discovery of antimutagenic agents is essential.
Harpagophytum procumbens
(HP) is traditionally used as anti-inflammatory and analgesic particularly against painful osteoarthritis. Harpagoside (HS), its major iridoid glycoside, is considered as the main active component.
Objective:
The aim of the present study was to evaluate the antimutagenic activity of HS and HP extracts against mutagenic activity of 1-NPy.
Materials and Methods:
The antimutagenic activity was investigated using the
in vitro
cytokinesis-block micronucleus assay in cultured human lymphocytes. Cells were exposed to HS or HP extracts before (pretreatment), during (co-treatment), and after (posttreatment) treatment with 1-NPy.
Results:
Results showed that HS significantly reduced the mutagenicity of 1-NPy in pretreatment and particularly in co-treatment, whereas all HP extracts significantly reduced the genotoxicity in the three protocols.
Conclusion:
These results suggested that HS was strongly involved in antimutagenic activity of HP extracts in co-treatment, but other components in HP extracts participated in this activity in pre- and post-treatment.
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Cytotoxic and apoptotic effects of
Ebenus boissieri
Barbey on human lung cancer cell line A549
p. 37
Esra Arslan Aydemir, Ece Simsek, Nilüfer Imir, Ramazan Süleyman Göktürk, Erdem Yesilada, Kayahan Fiskin
DOI
:10.4103/0973-1296.157679
PMID
:26109772
Background:
Fabaceae family members are known to possess preventive and therapeutic potentials against various types of cancers.
Objective:
The aim of this study was to investigate the cytotoxic and apoptotic effects of hydroalcoholic extracts from the aerial parts and roots of an endemic
Ebenus
species;
Ebenus boissieri
Barbey in human lung cancer cell line.
Materials
and
Methods:
After treatment with hydroalcoholic extracts cytotoxic activities of both extracts were measured by 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay, whereas caspase-3 activity, tumor necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) release were measured by enzyme linked immunosorbent assay.
Results:
According to
in vitro
assay results, the increase in all caspases activity suggested that extracts induce cells to undergo apoptosis. Especially, induction in caspase-3 activity was the most remarkable result of this study. Both aerial part and root extracts induced apoptosis by increasing caspase-3 activity, TNF-α and IFN-γ release. When compared to their relative controls, the concentrations of both TNF-α and IFN-γ in extract-treated groups were significantly and dose dependently exalted.
Conclusion:
Taken together, our results indicate that the hydroalcoholic extracts of
E. boissieri
can be considered as a source of new anti-apoptotic and therefore anti-carcinogenic agent.
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Antiproliferative efficacy of
Tabernaemontana divaricata
against HEP2 cell line and Vero cell line
p. 46
Arvind Kumar, S Selvakumar
DOI
:10.4103/0973-1296.157682
PMID
:26109773
Background:
Laryngeal cancer may also be called cancer of the larynx or laryngeal carcinoma. Conventional plants are a precious source of novel anticancer agents and are still in performance better role in health concern. The study was intended to estimation of the anticancer activity of the chloroformic extract of
Tabernaemontana
divaricata
on the human epidermoid larynx carcinoma cell line (Hep 2).
Materials
and
Methods:
The aerial parts (leaves, stem, and flowers) of
T.
divaricata
were tested for its inhibitory effect in 96 microplate formats against Hep 2 cell line. The anticancer activity of samples on Hep 2 and Vero was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and various enzymatic parameters like catalase, reduced glutathione (GSH), GSH peroxidase, and superoxide anion scavenging activity. Viable cells were determined by the absorbance at 540 nm. Measurements were performed, and the concentration required for a 50% inhibition of viability (IC50) was determined graphically. The effect of the samples on the proliferation of Hep 2 and Vero cells was expressed as the % cell viability.
Results:
The extract on Hep 2 cell line up to 7.8 μg/ml and that IC50 value on Hep 2 cell line was 112 μg whereas 94 μg for Vero cell line. Hence,
T.
divaricata
has lesser significant action on Vero cell line.
Conclusion:
Medicinal plant drug discovery continues to provide new and important leads against various pharmacological targets including cancer. Our results clearly indicate the anticancer property of the medicinal plant
T.
divaricata
against the human laryngeal carcinoma cell lines (Hep 2 cell line).
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Assay method for quality control and stability studies of a new anti-diabetic and anti-dyslipidemic flavone (S002-853)*
p. 53
Arshi Naqvi, Anuja Pandey, Varsha Gupta, Richa Malasoni, Akansha Srivastava, Rishi Ranjan Pandey, M Satyanarayana, Ram Pratap, Anil Kumar Dwivedi
DOI
:10.4103/0973-1296.157689
PMID
:26109774
Background:
Flavonoid-rich extract of the plant is long known for its anti-diabetic activities in traditional medicine. S002-853, a new flavone derivative synthesized by Central Drug Research Institute (CDRI) has been used for the present study.
Objectives:
The present study aimed at development of an assay method for quality control (QC) and stability studies of a new anti-diabetic and anti-dyslipidemic agent CDRI compound S002-853.
Materials and Methods:
A validated high-performance liquid chromatography analysis method for S002-853 was developed for in process QC and stability studies. The separation was achieved on a RP-C18 (25 cm × 0.4 cm, 5 μm, Phenomenex) at 240 nm with flow rate of 1.0 ml/min. This method was applied successfully in establishing forced degradation and drug-excipient testing protocols as per International Conference on Harmonization guidelines.
Results:
The result of estimation and stress testing studies indicated a high degree of selectivity of this method. S002-853 was most stable at pH 7 and under photolytic conditions. The temperature degradation pattern of S002-853 was found to follow the zero order degradation.
Conclusion:
The method described is easy and simple hence can be easily reproduced. This method can be very useful for bulk manufacture QC, and drug development process.
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Use of bergenin as an analytical marker for standardization of the polyherbal formulation containing
Saxifraga ligulata
p. 60
Hulikal Basavarajaiah Pushpalatha, Kumar Pramod, Rajendran Devanathan, Ramachandran Sundaram
DOI
:10.4103/0973-1296.157690
PMID
:26109776
Background:
Bergenin is the principle constituent of the well-known medicinal plant
Saxifraga ligulata
. Bergenin has anti-inflammatory, antipyretic, antiviral, immunostimulant, antihyperglycemic, and antioxidant properties. In this study, the presence of bergenin in
Saxifraga ligulata
and the formulations was identified using high performance thin layer chromatograpy fingerprinting technique.
Objective:
To develop a novel quantitative method for the estimation of bergenin using high performance liquid chromatography.
Materials and Methods:
The compound was separated, characterised and quantified using authentic reference standard. The method was validated per ICH guidelines for the parameters of accuracy, precision, linearity, limit of detection, limit of quantification and robustness.
Results:
The method was found to be accurate, linear (r2 = 0.998) and precise (%RSD <2%). The limits of detection (0.001%) and quantification (0.002%) were found to be suitable for detection and quantification of bergenin in commercial formulations.
Conclusion:
The developed methods are suitable for the quality control applications of Saxifraga ligulata containing formulations.
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A novel high-performance liquid chromatography-electron spray ionization-mass spectrometry method for simultaneous determination of guggulsterones, piperine and gallic acid in
Triphala guggulu
p. 66
Ganesh Muguli, PR Rao Vadaparthi, B Ramesh, Vishakante Gowda, Rangesh Paramesh, Atul N Jadhav, K Suresh Babu
DOI
:10.4103/0973-1296.157696
PMID
:26109777
"
Triphalaguggulu
0" is an important Ayurvedic formulation comprising of
Guggulu
, that is,
Commiphora
wightii (Arn.) Bhandari as a base wherein powdered fruits of
triphala
, that is,
Phyllanthus emblica
L.,
Terminalia bellirica
(Gaertn.) Roxb and
Terminalia chebula
Retz, along with powdered fruit of
Piper longum
L. are compounded. This polyherbal preparation has been strongly recommended in chronic inflammation, piles, and fistula. However, due to the complexity of compound formulation standardization of commercial products is challenging. In the present communication marker-based standardization of "
Triphalaguggulu
" preparation using gallic acid (for
triphala
), piperine (for
P. longum
L.) and guggulsterones (for guggulu) is reported. These compounds of diverse chemistry were successfully separated on a Waters HR-C18 column by isocratic elution with methanol and water (80:20 v/v) as mobile phase at the flow rate of 1.0 mL/min coupled with photodiode array detector. These optimal chromatographic conditions were used for simultaneous quantification of gallic acid, guggulsterones (E and Z) and piperine in commercial samples by high-performance liquid chromatography-electron spray ionization-mass spectrometry and method was validated as per ICH guidelines.
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Anti-lung cancer potential of pure esteric-glycoside condurangogenin A against nonsmall-cell lung cancer cells
in vitro
via p21/p53 mediated cell cycle modulation and DNA damage-induced apoptosis
p. 73
Sourav Sikdar, Avinaba Mukherjee, Anisur Rahman Khuda-Bukhsh
DOI
:10.4103/0973-1296.157698
PMID
:26109778
Background:
Marsdenia condurango
(condurango) is a tropical woody vine native to South America. Our earlier study was limited to evaluation of anti-cancer potentials of crude condurango extract and its glycoside-rich components
in vitro
on lung cancer.
Objective:
This study aims at evaluating the effect of the single isolated active ingredient condurangogenin A (ConA; C
32
H
42
O
7
) on A549, H522 and H460-nonsmall-cell lung cancer cells.
Materials
and
Methods:
ConA was isolated by column chromatography and analyzed by mass spectroscopy, Fourier transform infrared spectroscopy and proton-nuclear magnetic resonance. diphenyltetrazolium bromide assays were conducted on three cell-types using 6%-alcohol as control. Critical studies on cellular morphology, cell-cycle regulation, reactive oxygen species, mitochondrial membrane potential, and DNA-damage were made, and expressions of related signaling markers studied.
Results:
As IC
50
doses of ConA proved to be too high and toxic to both A549 and H522 cells, all experimental studies were carried out on H460 cells with the IC
50
dose (32 μg/ml − 24 h). Cellular morphology revealed typical apoptotic features after ConA treatment. At early treatment hours (2 h-12 h), maximum cells were arrested at G0/G1 phase that could be correlated with reduced level of cyclin D1-CDK with p21 up-regulation. At 18 h − 24 h, sub G0/G1 cell population was increased gradually, as revealed from cytochrome-c release and caspase-3 activation, further confirming the apoptosis-inducing ability of ConA at later phases. Gradual increase of TUNEL-positive cells with significant modulation of mitochondria-dependent apoptotic markers at longer time-points would establish apoptosis-induction property of ConA, indicating its potential as a strong candidate for anti-cancer drug formulation.
Conclusion:
Further studies are warranted against other types of cancer cells and animal models before its possible human use.
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Protective effect of young green barley leaf (
Hordeum vulgare
L.) on restraint stress-induced decrease in hippocampal brain-derived neurotrophic factor in mice
p. 86
Katsunori Yamaura, Riho Tanaka, Yuanyuan Bi, Hideki Fukata, Nobuo Oishi, Hiromi Sato, Chisato Mori, Koichi Ueno
DOI
:10.4103/0973-1296.157702
PMID
:26109779
Background:
Many health experts support the hypothesis that stressful lifestyles are the leading cause of illness, like depression. Therefore, from the standpoint of preventive medicine, it is important to reduce stress. Young green barley leaves are a good natural source of vitamins and minerals, and their juice is widely consumed as a functional food for health reasons in Japan. This study investigated the protective effect of young green barley leaves for stress control.
Materials and
Methods:
ICR outbred mice were exposed to 3-h sessions of restraint stress. Young green barley leaves (400 and 1,000 mg/kg) were administered orally 1 h before the sessions for 5 days. To analyze voluntary behavior, wheel-running activity was monitored during the dark period. Brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) expression in the whole hippocampus was measured by real-time quantitative polymerase chain reaction.
Results:
Restraint stress resulted in a significant decrease in voluntary wheel-running behavior, but this decrease was ameliorated by the administration of young green barley leaves. The leaves also enhanced the decreased levels of BDNF mRNA induced by restraint stress; in particular, a significant protective effect was shown in the exon IV variant as compared to vehicle control mice.
Conclusion:
The findings suggest that young green barley leaves have potent anti-stress properties, as evidenced by preventing decreases in the levels of voluntary wheel-running activity and hippocampal BDNF mRNA in response to restraint stress. Our findings support the possibility that supplementation with young green barley leaves might be beneficial for preventing stress-related psychiatric disorders like depression.
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Toxicological and biochemical studies on
Schinus terebinthifolius
concerning its curative and hepatoprotective effects against carbon tetrachloride-induced liver injury
p. 93
Rania H Abdou, Sherif Y Saleh, Waleed F Khalil
DOI
:10.4103/0973-1296.157705
PMID
:26109780
Background:
Recently, many efforts have been made to discover new products of natural origin which can limit the xenobiotic-induced hepatic injury. Carbon tetrachloride (CCl
4
) is a highly toxic chemical that is widely used to study hepatotoxicity in animal models.
Objective:
The present study was conducted to investigate the curative and protective effects of
Schinus terbenthifolius
ethanolic extract against CCl
4
-induced acute hepatotoxicity in rats.
Materials
and
Methods:
S. terbenthifolius
extract was orally administered in a dose of 350 mg dried extract/kg b.wt. before and after intoxication with CCl
4
for curative and protective experiments, respectively. A group of hepatotoxicity indicative enzymes, oxidant-antioxidant capacity, DNA oxidation, and apoptosis markers were measured.
Results:
CCl
4
increased liver enzyme leakage, oxidative stress, hepatic apoptosis, DNA oxidation, and inflammatory markers. Administration of
S. terebinthifolius
, either before or after CCl
4
intoxication, significantly decreased elevated serum liver enzymes and reinstated the antioxidant capacity. Interestingly,
S. terebinthifolius
extract inhibited hepatocyte apoptosis as revealed by approximately 20 times down-regulation in caspase-3 expression when compared to CCl
4
untreated group. On the other hand, there was neither protective nor curative effect of
S. terebinthifolius
against DNA damage caused by CCl
4
.
Conclusion:
The present study suggests that
S. terebinthifolius
extract could be a substantially promising hepatoprotective agent against CCl
4
toxic effects and may be against other hepatotoxic chemical or drugs.
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Flavonoid profile, antioxidant and cytotoxic activity of different extracts from Algerian
Rhamnus alaternus
L. bark
p. 102
Soulef Boussahel, Antonio Speciale, Saliha Dahamna, Yacine Amar, Irene Bonaccorsi, Francesco Cacciola, Francesco Cimino, Paola Donato, Guido Ferlazzo, Daoud Harzallah, Mariateresa Cristani
DOI
:10.4103/0973-1296.157707
PMID
:26109754
Background:
Rhamnus alaternus (Rhamnaceae) L. has been traditionally used for treatment of many diseases.
Objective:
In this study, we determined the antioxidant/free radical scavenger properties, the flavonoid profile and the cytotoxicity of aqueous and methanolic extracts obtained by maceration from Algerian R. alaternus bark, like also of aqueous extract prepared by decoction according to the traditional method. This to estimate the usefulness of the drug traditional preparation and compare it with those made in the laboratory.
Materials and Methods:
The antioxidant activity of the extracts was evaluated using five different redox-based assays, all involving one redox reaction with the oxidant. High-performance liquid chromatography/diode array detection/electrospray ionization mass spectrometry analysis was used to identify and quantify the fl avonoids content. Cytotoxicity on human monocytic leukemia cells (U937) was also carried out.
Results:
All the extracts tested showed a good antioxidant/free radical scavenger activity and a similar flavonoid fingerprint. However, the methanolic one presented the best antioxidant activity that can be due to the highest flavonoid amount and significantly reduced the proliferation of leukemia cells. The results confirm that the extract prepared by decoction contains efficient antioxidant compounds and this justifies in part the therapeutic and preventive usefulness. Moreover, the methanolic extract exerted excellent cytotoxicity on U937 that could be attributed to kaempferol and rhamnocitrin glycosides.
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Inhibitory potential of some Romanian medicinal plants against enzymes linked to neurodegenerative diseases and their antioxidant activity
p. 110
Gabriela Paun, Elena Neagu, Camelia Albu, Gabriel Lucian Radu
DOI
:10.4103/0973-1296.157709
PMID
:26109755
Context:
Eryngium planum, Geum urbanum
and
Cnicus benedictus
plants are an endemic botanical from the Romanian used in folk medicine.
Objective:
The extracts from three Romanian medicinal plants were investigated for their possible neuroprotective potential.
Materials and Methods:
Within this study,
in vitro
neuroprotective activity of the extracts of
E. planum, G. urbanum,
and
C. benedictus
plants were investigated via inhibition of acetylcholinesterase (AChE) and tyrosinase (TYR). Total content of phenolics, flavonoids, and proanthocyanidins, high-performance liquid chromatography profile of the main phenolic compounds and antioxidant activity were also determined.
Results:
Among the tested extracts, the best inhibition of AChE (88.76 ± 5.2%) and TYR (88.5 ± 5.2%) was caused by
C. benedictus
ethanol (EtOH) extract. The
G. urbanum
extracts exerted remarkable scavenging effect against 2,2-diphenyl-1-picrylhydrazyl (IC
50
, 7.8 ± 0.5 μg/mL aqueous extract, and IC
50
, 1.3 ± 0.1 μg/mL EtOH extract, respectively) and reducing power, whereas the EtOH extract of
C. benedictus
showed high scavenging activity (IC
50
, 0.609 ± 0.04 mg/mL), also.
Conclusion:
According to our knowledge, this is the first study that demonstrates
in vitro
neuroprotective effects of
E. planum, G. urbanum
and
C. benedictus
.
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Experimental treatment of breast cancer-bearing BALB/c mice by artemisinin and transferrin-loaded magnetic nanoliposomes
p. 117
Amir Gharib, Zohreh Faezizadeh, Seyed Ali Reza Mesbah-Namin, Ramin Saravani
DOI
:10.4103/0973-1296.157710
PMID
:26109756
Background:
The combination of artemisinin and transferrin exhibits versatile anticancer activities. In previous, we successfully prepared artemisinin and transferrin-loaded magnetic nanoliposomes and evaluated their anti-proliferative activity against MCF-7 and MDA-MB-231 cell lines
in vitro
. In this study, we investigate the
in vivo
anti-breast cancer activity of artemisinin and transferrin-loaded magnetic nanoliposome against breast transplanted tumors in BALB/c mice model.
Materials
and
Methods:
Artemisinin and transferrin-loaded magnetic nanoliposomes were prepared and characterized for some physiochemical properties. Pieces of tumor tissue from the breast cancer-bearing BALB/c mice were transplanted subcutaneously to the syngeneic female BALB/c mice. In the presence of the external magnet that placed at the breast tumor site, the tissue distribution and tumor-suppressing effects of prepared nanoliposomes on tumor growth was evaluated.
Results:
The prepared nanoliposomes have fine spherical shape, rough surface, nano-sized diameter and magnetic properties. At 2 h after treatment, the intravenous administration of artemisinin and transferrin-loaded magnetic nanoliposomes followed using the magnetic field approximately produced 10- and 5.5-fold higher levels of artemisinin and transferrin in the tumors, respectively, compared with free artemisinin and transferrin. Moreover, in the presence of an external magnetic field, the prepared nanoliposomes could significantly induce apoptosis in the mice breast cancer cells as well as could reduce tumor volume in tumorized mice at 15 days after treatment.
Conclusion:
The data suggested that the artemisinin and transferrin-loaded magnetic nanoliposomes would be a good choice for the breast tumor-targeted therapy, due to its high targeting efficiency.
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In silico
analysis and molecular docking studies of potential angiotensin-converting enzyme inhibitor using quercetin glycosides
p. 123
Syed Aun Muhammad, Nighat Fatima
DOI
:10.4103/0973-1296.157712
PMID
:26109757
The purpose of this study was to analyze the inhibitory action of quercetin glycosides by computational docking studies. For this, natural metabolite quercetin glycosides isolated from buckwheat and onions were used as ligand for molecular interaction. The crystallographic structure of molecular target angiotensin-converting enzyme (ACE) (peptidyl-dipeptidase A) was obtained from PDB database (PDB ID: 1O86). Enalapril, a well-known brand of ACE inhibitor was taken as the standard for comparative analysis. Computational docking analysis was performed using PyRx, AutoDock Vina option based on scoring functions. The quercetin showed optimum binding affinity with a molecular target (angiotensin-converting-enzyme) with the binding energy of −8.5 kcal/mol as compared to the standard (−7.0 kcal/mol). These results indicated that quercetin glycosides could be one of the potential ligands to treat hypertension, myocardial infarction, and congestive heart failure.
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Development and extraction optimization of baicalein and pinostrobin from
Scutellaria violacea
through response surface methodology
p. 127
Shankar Subramaniam, Ravikumar Raju, Anbumathi Palanisamy, Aravind Sivasubramanian
DOI
:10.4103/0973-1296.157714
PMID
:26109758
Objective:
To develop a process that involves optimization of the amount of baicalein and pinostrobin from the hydro-methanolic extract of the leaves of
Scutellaria violacea
by response surface methodology (RSM).
Materials and Methods:
The combinatorial influence of various extraction parameters on the extraction yield was investigated by adopting Box-Behnken experimental design. Preliminary experiments carried out based on the traditional one variable at a time optimization revealed four such operational parameters to play a crucial role by influencing the yield. These four process parameters at three levels were considered to obtain the Box-Behnken experimental design.
Results:
RSM based model fitted to the resulting experimental data suggested that 52.3% methanol/water, 12.46:1 solvent-solid ratio, 285 rpm agitation and 6.07 h of extraction time are the optimal conditions which yielded a maximized amount of baicalein and pinostrobin of 2.9 and 4.05 mg/g DM. Analysis of variance revealed a high correlation coefficient (
R
2
= 0.999 for baicalein and 0.994 for pinostrobin), signifying a good fit between the regression model (second order) and the experimental observations.
Conclusion:
The present study signifies that both the metabolites have been extracted from
S. violacea
for the first time. Further, this study developed an optimized extraction procedure to obtain maximum yield of the metabolites, which is unique and better than conventional extraction methodology. The operational parameters under optimized conditions accounts for the lowest cost in extraction process thus, providing an efficient, rapid and cost-effective method for isolation and scale up of these commercially vital flavonoids.
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Methanol extract of wheatgrass induces G1 cell cycle arrest in a p53-dependent manner and down regulates the expression of cyclin D1 in human laryngeal cancer cells-an
in vitro
and
in silico
approach
p. 139
Garima Shakya, Sangeetha Balasubramanian, Rukkumani Rajagopalan
DOI
:10.4103/0973-1296.157715
PMID
:26109759
Background:
Deregulation of cell cycle has been implicated in the malignancy of cancer. Since many years investigation on the traditional herbs has been the focus to develop novel and effective drug for cancer remedies. Wheatgrass is a medicinal plant, used in folk medicine to cure various diseases. The present study was undertaken to gain insights into antiproliferative effect of methanol extract of wheatgrass.
Materials
and
Methods:
Cell viability was assessed via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Lactate Dehydrogenase assays. Cell cycle was analyzed by flow cytometry. Western blot was performed to determine the p53 and cyclin D1 levels.
In silico
docking interaction of the 14 active components (identified by high-performance liquid chromatography/gas chromatography-mass spectroscopy) of the methanol extract was tested with cyclin D1 (Protein Data Bank ID: 2W96) and compared with the reference cyclin D1/Cdk4 inhibitor.
Results:
Methanol extract of wheatgrass effectively reduced the cell viability. The cell cycle analysis showed that the extract treatment caused G
1
arrest. The level of cyclin D1 was decreased, whereas p53 level was increased. Molecular docking studies revealed interaction of seven active compounds of the extract with the vital residues (Lys112/Glu141) of cyclin D1.
Conclusion:
These findings indicate that the methanol extract of wheatgrass inhibits human laryngeal cancer cell proliferation via cell cycle G
1
arrest and p53 induction. The seven active compounds of the extract were also found to be directly involved in the inhibition of cyclin D1/Cdk4 binding, thus inhibiting the cell proliferation.
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The dose dependent
in vitro
responses of MCF-7 and MDA-MB-231 cell lines to extracts of
Vatica diospyroides
symington type SS fruit include effects on mode of cell death
p. 148
Theera Srisawat, Yaowapa Sukpondma, Potchanapond Graidist, Siriphon Chimplee, Kanyanatt Kanokwiroon
DOI
:10.4103/0973-1296.157718
PMID
:26109760
Background:
Vatica diospyroides
type LS is a known source of valuable compounds for cancer treatment, however, in contrast little is known about therapeutic efficacy of type SS.
Objective:
This study focused on
in vitro
cytotoxicity of these fruit extracts, and the cell death mode they induce in breast cancer cells.
Materials and Methods:
Acetone extracts of fruit were tested for cytotoxicity against MCF-7 and MDA-MB-231 cell lines. The apoptosis and necrosis of these cells were quantified by fluorescence activated cell sorter (FACS) and western blot analyses.
Results:
After 72 h of treatment, the 50% growth inhibition concentrations (IC
50
) levels were 16.21 ± 0.13 µg/mL against MCF-7 and 30.0 ± 4.30 µg/mL against MDA-MB-231, indicating high and moderate cytotoxicity, respectively. From the FACS results, we estimate that the cotyledon extract at half IC
50
produced 11.7% dead MCF-7 cells via apoptosis, whereas another concentrations both apoptosis and necrosis modes co-existed in a dose-dependent manner. In MDA-MB-231 cell line, only the apoptosis was induced by the pericarp extract in a dose-dependent manner. With the extracts at half IC
50
concentration, in both cells, the expression of p21 decreased while that of Bax increased within 12-48 h of dosing, confirming apoptosis induced by time-dependent responses. Apoptosis dependent on p53 was found in MCF-7, whereas the mutant p53 of MDA-MB-231 cells was expressed.
Conclusion:
The results indicate that fruit extracts of
V. diospyroides
have cytotoxic effects against MCF-7 and MDA-MB-231 cells via apoptosis pathway in a dose-dependent manner. This suggests that the extracts could provide active ingredients for the development, targeting breast cancer therapy.
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Cytotoxic activity of
Macrosolen parasiticus
(L.) Danser on the growth of breast cancer cell line (MCF-7)
p. 156
Vijay Kumar Sodde, Richard Lobo, Nimmy Kumar, Rajalekshmi Maheshwari, CS Shreedhara
DOI
:10.4103/0973-1296.157719
PMID
:26109761
Background:
Macrosolen parasiticus
(L.) Danser belonging to
Loranthaceaea
(mistletoe family) is a parasitic plant that grows on different host plants such as mango, jack fruit, peepal, neem tree, etc., This study was aimed to investigate the anti-cancer activity of methanolic and aqueous extract of stem of
M. parasiticus
.
Objectives:
To investigate the
in vitro
cytotoxic potential of the methanolic and aqueous extracts from stems of
M. parasiticus
against MCF-7 breast cancer cells by brine shrimp lethality (BSL) bioassay, MTT assay and sulforhodamine B (SRB) assay.
Materials and Methods:
The extracts were tested in human breast cancer cell lines
in vitro
for percentage cytotoxicity, apoptosis by acridine orange/ethidium bromide staining, LD
50
and IC
50
values after treatment with
M. parasiticus
extracts.
Results:
In BSL bioassay, aqueous extract showed more significant (
P
< 0.01) cytotoxicity with LD
50
82.79 ± 2.67 μg/mL as compared to methanolic extract with LD
50
125 ± 3.04 μg/mL. The methanolic extract of
M. parasiticus
showed IC
50
97.33 ± 3.75 μg/mL (MTT) (
P
< 0.05) and 94.58 ± 3.84 μg/mL (SRB) (
P
< 0.01) assays against MCF-7. The aqueous extract of
M. parasiticus
demonstrated higher activity with IC
50
59.33 ± 3.3 μg/mL (MTT) (
P
< 0.01) and 51.9 ± 1.87 μg/mL (SRB) (
P
< 0.01) assays, after 48 h of exposure and thus showed significant dose-dependent cytotoxic activity.
Conclusion:
The finding demonstrated that both extracts of
M. parasiticus
showed significant cytotoxic activity, however aqueous extract demonstrated higher activity against MCF-7 breast cancer cells.
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Effects of the extract and glycoalkaloids of
Solanum lycocarpum
St. Hill on
Giardia lamblia
trophozoites
p. 161
Gilmarcio Z Martins, Raquel R. D. Moreira, Cleopatra S Planeta, Adélia E Almeida, Jairo K Bastos, Lígia Salgueiro, Carlos Cavaleiro, Maria do Céu Sousa
DOI
:10.4103/0973-1296.157721
PMID
:26109762
Background:
Solanum lycocarpum
has great importance for food and medicinal traditional use. Recently, it was also evidenced that extracts of
S. lycocarpum
St. Hill (Solanaceae) and its glycoalkaloids, solamargine (Sg) and solasonine (Sn), are active against flagellated protozoa.
Objective:
The aim was to assess the effects of the extract of
S. lycocarpum
and its glycoalkaloids, Sn, and Sg, on
Giardia lamblia
trophozoites.
Materials
and
Methods:
A crude extract (96% ethanol) (EB) of fruits of
S. lycocarpum
was prepared and fractionated by partition with 40% ethanol and
n
-hexane: Ethyl acetate. Glycoalkaloids, Sn, and Sg were recognized in the ethanol fraction (EF) and further isolated by column chromatography. EB, EF, the isolated Sn and Sg and a mixture (1:1) of both glycoalkaloids were tested on cultures of
G. lamblia
trophozoites and macrophages.
Results:
EB, EF and glycoalkaloids of
S. lycocarpum
showed activity against
Giardia
(95.0 < Inhibitory concentration 50 [IC
50
] ≤120.3 μg/mL). The mixture of glycoalkaloids (1:1) was more active (IC
50
= 13.23 μg/mL) than each one individually, suggesting a synergic effect. Moreover, the mixture is nontoxic to macrophage cells.
Conclusion:
Results are optimistic concerning the anti-
Giardia
potential of the mixture Sn + Sg. Further studies,
in vitro
and
in vivo
, will be required to consolidate the usefulness of the mixture of Sn + Sg in view of a new therapeutic strategy for giardiasis.
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Development of response surface methodology for optimization of extraction parameters and quantitative estimation of embelin from
Embelia ribes
Burm by high performance liquid chromatography
p. 166
Md. Shamsir Alam, Zoheir A Damanhouri, Aftab Ahmad, Lubna Abidin, Mohd Amir, Mohd Aqil, Shah Alam Khan, Mohd Mujeeb
DOI
:10.4103/0973-1296.157722
PMID
:26109763
Background:
Embelia ribes
Burm is widely used medicinal plant for the treatment of different types of disorders in the Indian traditional systems of medicine.
Objective:
The present work was aimed to optimize the extraction parameters of embelin from
E. ribes
fruits and also to quantify embelin content in different extracts of the plant.
Materials and Methods:
Optimization of extraction parameters such as solvent: drug ratio, temperature and time were carried out by response surface methodology (RSM). Quantitative estimation of embelin in different extracts of
E. ribes
fruits was done through high performance liquid chromatography.
Results:
The optimal conditions determined for extraction of embelin through RSM were; extraction time (27.50 min), extraction temperature 45°C and solvent: drug ratio (8:1). Under the optimized conditions, the embelin yield (32.71%) was equitable to the expected yield (31.07%,
P
> 0.05). These results showed that the developed model is satisfactory and suitable for the extraction process of embelin. The analysis of variance showed a high goodness of model fit and the accomplishment of the RSM method for improving embelin extraction from the fruits of
E. ribes
.
Conclusion:
It is concluded that this may be a useful method for the extraction and quantitative estimation of embelin from the fruits of
E. ribes
.
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Hepatorenal protective effect of Antistax
®
against chemically-induced toxicity
p. 173
Atallah F Ahmed, Hanan M Al-Yousef, Jawaher H Al-Qahtani, Mansour S Al-Said, AbdelKader E Ashour, Mohammed Al-Sohaibani, Syed Rafatullah
DOI
:10.4103/0973-1296.157726
PMID
:26109764
Background:
Antioxidant natural products and chemoprevention are considered nowadays as an effective approach against health various disorders and diseases induced by oxidative stress or free radicals.
Objective:
The aim of this study was to assess the hepato- and nephroprotective activity of a standardized red vine leaf aqueous extract AS195 (Antistax
®
).
Methods:
The protective activity of AS195 (100 mg/kg) was investigated on carbon tetrachloride (CCl
4
)-intoxicated rats in comparison with silymarin. The flavonoid/proanthocyanidin nature of AS195 was identified by phytochemical and nuclear magnetic resonance (NMR) analyses, while its total phenol/proanthocyanidin/flavonoid content and antioxidant activity were determined by Folin-Ciocalteau, vanillin-sulfuric acid, AlCl
3
, and 2,2-diphenyl-2-picrylhydrazyl radical scavenging assays, respectively.
Results:
Relative to the control CCl
4
-intoxicated group, pretreatment with AS195 could significantly suppressed the elevated serum levels of alanine aminotransferase, alkaline phosphatase, γ-glutamyl transferase, total cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, triglycerides, bilirubin, creatinine, uric acid, and calcium, whereas it significantly increased the diminished serum levels of high-density lipoprotein cholesterol, albumin and total protein. Moreover, AS195 significantly decreased malondialdehyde formation in the tissues of liver and kidney, whereas it significantly elevated and nonprotein sulfhydryl groups, compared with the intoxicated control. The improvement in biochemical parameters by AS195 was obviously observed and further confirmed by restoration of normal histological features in the two organs.
Conclusions:
The results of the present study revealed the capacity of AS195 to enhance the recovery from xenobiotic-induced hepatorenal toxicity initiated by free radicals.
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Neuroprotective effects of
Nigella sativa
extracts during germination on central nervous system
p. 182
Mohammad Hayatul Islam, Iffat Zareen Ahmad, Mohammad Tariq Salman
DOI
:10.4103/0973-1296.157729
PMID
:26109765
Background:
Nigella sativa
Linn. which has many acclaimed medicinal properties is an indigenous herbaceous plant and belongs to the
Ranunculaceae
family, which grows in countries bordering the Mediterranean Sea, Pakistan and India.
Objective:
This study was designed to investigate the effects of
N. sativa
seed extracts of different germination phases on the central nervous system (CNS) responses in experimental animals.
Materials and Methods:
Anxiolytic, locomotor activity of extracts (1 g/kg of body weight) was evaluated in both stressed and unstressed animal models and antiepileptic effect was evaluated by maximal electroshock seizure model keeping diazepam (20 mg/kg) as a positive control. Antidepressant effect was evaluated by forced swim test and tail suspension test keeping imipramine (15 mg/kg) as a positive control.
Results:
All tested extracts of
N. sativa
during different phases of germination (especially 5
th
day germination phase) showed significant (
P
< 0.001) anxiolytic effect in comparison to control. Diazepam reduced locomotor activity in control (unstressed) rats but did not show affect in stressed rats while
N. sativa
extracts from germination phases significantly (
P
< 0.001) reduced locomotor activity in unstressed as well as stressed animals. All the extracts of
N. sativa
from different germination phases exhibited significant (
P
< 0.001) reduction in various phases of epileptic seizure on comparison with the reference standard (diazepam). During antidepressant test,
N. sativa
extracts exhibited a slight reduction in the immobility of rats.
Conclusion:
During germination, especially in 5
th
day germination extract,
N. sativa
showed significant CNS depressant activity as compared to whole seeds that possibly may be due higher content of secondary metabolites produced during germination.
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The potential of selected Australian medicinal plants with anti-Proteus activity for the treatment and prevention of rheumatoid arthritis
p. 190
IE Cock, V Winnett, J Sirdaarta, B Matthews
DOI
:10.4103/0973-1296.157734
PMID
:26109767
Background:
A wide variety of herbal medicines are used in indigenous Australian traditional medicinal systems to treat rheumatoid arthritis (RA) and inflammation. The current study was undertaken to test the ability of a panel of Australian plants with a history of the ethnobotanical usage in the treatment of inflammation for the ability to block the microbial trigger of RA.
Materials and Methods:
One hundred and six extracts from 40 plant species were investigated for the ability to inhibit the growth of the bacterial trigger of RA (Proteus mirabilis). The extracts were tested for toxicity in the Artemia nauplii bioassay. The most potent inhibitor of P. mirabilis growth was further analyzed by reversed-phase high performance liquid chromatography (RP-HPLC) coupled to high accuracy time-of-flight (TOF) mass spectroscopy.
Results:
Sixty-five of the 106 extracts tested (61.3%) inhibited the growth of P. The
Aleurites moluccanus, Datura leichardtii, Eucalyptus major, Leptospermum bracteata, L. juniperium, Macadamia integriflora
nut,
Melaleuca alternifolia, Melaleuca quinquenervia, Petalostigma pubescens, P. triloculorae, P. augustifolium, Scaevola spinescens, Syzygium australe, and Tasmannia lanceolata
extracts were determined to be the most effective inhibitors of
P. mirabilis
growth, with minimum inhibitory concentration (MIC) values generally significantly below 1000 μg/ml.
T.
lanceolata
fruit extracts were the most effective
P.
mirabilis
growth inhibitors, with a MIC values of 11 and 126 μg/ml for the methanolic and aqueous extracts, respectively. Subsequent analysis of the
T.
lanceolata
fruit extracts by RP-HPLC coupled to high-resolution TOF mass spectroscopy failed to detect resveratrol in either
T. lanceolata
fruit extract. However, the resveratrol glycoside piceid and 2 combretastatin stilbenes (A-1 and A-4) were detected in both T. lanceolata fruit extracts. With the exception of the Eucalyptus and Syzygium extracts, all extracts exhibiting Proteus inhibitory activity were also shown to be nontoxic, or of low toxicity in the Artemia nauplii bioassay.
Conclusions:
The low toxicity of these extracts and their inhibitory bioactivity against Proteus spp. indicate their potential in blocking the onset of rheumatoid arthritis.
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Protective role of Kalpaamruthaa in type II diabetes mellitus-induced cardiovascular disease through the modulation of protease-activated receptor-1
p. 209
Raja Latha, Palanivelu Shanthi, Panchanadham Sachdanandam
DOI
:10.4103/0973-1296.157739
PMID
:26109768
Background:
Kalpaamruthaa (KA) is a formulatory herbal preparation has beneficial antioxidant, anti-apoptotic and anti-inflammatory properties against cardiovascular damage (CVD).
Objective:
The present study was undertaken to investigate the protective role of KA in type II diabetes mellitus-induced CVD through the modulation of protease-activated receptor-1 (PAR1).
Materials and Methods:
CVD was developed in 8 weeks after type II diabetes mellitus induction with high fat diet (2 weeks) and low dose of streptozotocin (2 × 35 mg/kg b.w. i.p. in 24 h interval). CVD-induced rats treated with KA (200 mg/kg b.w. in 0.5 ml of olive oil) orally for 4 weeks.
Results:
KA increased the activities of enzymatic antioxidants and the levels of non-enzymatic antioxidants in pancreas of CVD-induced rats. KA effectively reduced the lipid peroxides and carbonyl content in the pancreas of CVD-induced rats. KA reduced cellular damage by ameliorating the activities of marker enzymes in plasma, heart and liver. The protective nature of KA was further evidenced by histological observation in pancreas. Further, KA reduced CVD by decreasing the expression of PAR1 in heart.
Conclusion:
This study exhibits the defending role of KA in type II diabetes mellitus-induced CVD through altering PAR1.
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Pharmacological and phytochemical evaluation of Ocimum sanctum root extracts for its antiinflammatory, analgesic and antipyretic activities
p. 217
Anant Kumar, Karishma Agarwal, Anil Kumar Maurya, Karuna Shanker, Umme Bushra, Sudeep Tandon, Dnyaneshwar U Bawankule
DOI
:10.4103/0973-1296.157743
PMID
:26109769
Background:
Long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) increases risk of having a range of gastrointestinal problems. Therefore, new anti-inflammatory, analgesic, antipyretic drugs having lesser side effects are being searched all over the world as alternatives to NSAIDs.
Aims:
To evaluate the anti-inflammatory, analgesic and antipyretic profile of
Ocimum sanctum
root extracts.
Materials and Methods:
Anti-inflammatory profile of hexane (STH), chloroform (STC), ethyl acetate (STE), butanol (STB) and water (STW) extracts of OS was carried out by using carrageenan induced paw edema. STE a most active extract was further validated in dose dependent manner for anti-inflammatory, analgesic and antipyretic activity as well as oral toxicity profile in small laboratory animals. Identification of bioactives flux and chemical signature of most active fraction STE was developed by using the high-performance liquid chromatography fingerprinting.
Results:
An ethyl acetate fraction (STE) exhibit most potent anti-inflammatory activity followed by STB, STW, STC and STH. Dose response study of STE showed anti-inflammatory, analgesic and anti-pyretic potential in dose-dependent manner without any toxic effect at dose 2000 mg/kg. Chemical fingerprint revealed the presence of flavanoids.
Conclusions:
The present research revealed that STE possess anti-inflammatory, analgesic and anti-pyretic properties. However, future research is advocated to evaluate the pharmacological properties of isolated bioactive compounds.
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Phytoecdysteroids of the East Asian
Caryophyllaceae
p. 225
Elena Novozhilova, Viacheslav Rybin, Petr Gorovoy, Irina Gavrilenko, Roman Doudkin
DOI
:10.4103/0973-1296.157746
PMID
:26109770
Background:
Occurrence of integristerone A (1), 20-hydroxyecdysone (2), ecdysone (3), 2-deoxy-20-hydroxyecdysone (4) has been analyzed in 64 species of the East Asian
Caryophyllaceae.
Materials
and
Methods:
Ecdysteroid content was determinate by high-performance liquid chromatography (HPLC). HPLC with a high-resolution mass spectrometry was performed on Shimadzu LCMS-IT-TOF (Japan) system equipped with a LC-20A Prominence liquid chromatograph, a photodiode array detector SPD-M20A and ion-trap/time-of-flight mass spectrometer.
Results:
New sources of phytoecdysteroids:
Melandrium
sachalinense
and
Melandrium
firmum
have been revealed. It is the 1
st
time that two has been identified in
M.
sachalinense
and
M.
firmum
; 1 in the species:
Lychnis
fulgens,
Silene
repens,
Silene
foliosa,
Silene
stenophylla,
Silene
jenisseensis
and
M.
sachalinense
; 3 in
Lychnis
cognata
; 4 in
L.
fulgens,
S.
stenophylla
and
S.
jenisseensis
(the tribe
Lychnideae
, the subfamily
Caryophylloideae
). Ecdysteroid-negative taxa are
Spergularia
rubra
of the tribe
Sperguleae
; species of the genera
Minuartia,
Honckenya,
Eremogone,
Arenaria,
Moehringia,
Pseudostellaria,
Fimbripetalum,
Stellaria
and
Cerastium
of the tribe
Alsineae
;
Scleranthus
annuus
of the tribe
Sclerantheae
, as well as the East Asian representatives of the genera
Gypsophila,
Psammophiliela
,
Dianthus
and
Saponaria
of the tribe
Diantheae
;
Oberna
and
Agrostemma
of the tribe
Lychnideae
.
Conclusion:
This investigation shows the most promising sources of ecdysteriods are species of genera
Silene
and
Lychnis
.
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