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Table of Contents
January-March 2014
Volume 10 | Issue 37
Page Nos. 1-88
Online since Friday, February 7, 2014
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ORIGINAL ARTICLES
Crude ethyl acetate extract of marine microalga,
Chaetoceros calcitrans
, induces Apoptosis in MDA-MB-231 breast cancer cells
p. 1
Su Hua Goh, Noorjahan Banu Mohamed Alitheen, Fatimah Md Yusoff, Swee Keong Yap, Su Peng Loh
DOI
:10.4103/0973-1296.126650
PMID
:24696543
Background:
Marine brown diatom
Chaetoceros
calcitrans
and green microalga
Nannochloropsis
oculata
are beneficial materials for various applications in the food, nutraceutical, pharmaceutical and cosmeceutical industries.
Objective:
This study investigated cytotoxicity of different crude solvent extracts from
C.
calcitrans
and
N.
oculata
against various cancer cell lines.
Materials
and
Methods:
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was carried out to screen the cytotoxic effects of hexane (Hex), dichloromethane (DCM), ethyl acetate, and methanol extract from
C.
calcitrans
and
N.
oculata
toward various cancer cell lines. Flow cytometry cell cycle was used to determine the cell cycle arrest while the mode of cell death was investigated through acridine orange/propidium iodide (AOPI) staining, Annexin V-Fluorescein Isothiocyanate (FITC) and Terminal deoxynucleotidyl transferase-mediated d-UTP Nick End Labeling (TUNEL) assays. Expression profile of apoptotic and proliferative-related genes was then determined using the multiplex gene expression profiler (GeXP).
Results:
Crude ethyl acetate (CEA) extract of
C.
calcitrans
inhibited growth of MDA-MB-231 cells, with IC
50
of 60 μg/mL after 72 h of treatment. Further studies were conducted to determine the mode of cell death at various concentrations of this extract: 30, 60 and 120 μg/mL. The mode of cell death was mainly apoptosis as shown through apoptosis determination test. The expression data from GeXP showed that caspase-4 was upregulated while B-cell leukemia/lymphoma 2(Bcl-2) was down regulated. Thus, caspase-4 induction endoplasmic reticulum death pathway is believed to be one of the mechanisms underlying the induction of apoptosis while Bcl-2 induced S and G2/M cell cycle phase arrest in MDA-MB-231 cells
.
Conclusion:
CEA extract of
C.
calcitrans
showed the highest cytotoxicity on MDA-MB-231 via apoptosis.
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Effect of various absorption enhancers based on tight junctions on the intestinal absorption of forsythoside A in Shuang-Huang-Lian, application to its antivirus activity
p. 9
Wei Zhou, Xuan Xuan Zhu, Ai Ling Yin, Bao Chang Cai, Hai Dan Wang, Liuqing Di, Jin Jun Shan
DOI
:10.4103/0973-1296.126651
PMID
:24695554
Background:
Forsythoside A (FTA), one of the main active ingredients in Shuang-Huang-Lian (SHL), possesses strong antibacterial, antioxidant and antiviral effects, and its pharmacological effects was higher than that of other ingredients, but the absolute bioavailability orally was approximately 0.72%, which was significantly low, influencing clinical efficacies of its oral preparations seriously.
Materials and Methods:
In vitro
Caco-2 cell and
in vivo
pharmacokinetics study were simultaneously performed to investigate the effects of absorption enhancers based on tight junctions: sodium caprate and water-soluble chitosan on the intestinal absorption of FTA, and the eventual mucosal epithelial damage resulted from absorption enhancers was evaluated by MTT test and morphology observation, respectively. The pharmacological effects such as antivirus activity improvement by absorption enhancers were verified by MDCK damage inhibition rate after influenza virus propagation.
Results:
The observations from
in vitro
Caco-2 cell showed that the absorption of FTA in SHL could be improved by absorption enhancers. Meanwhile, the absorption enhancing effect of water-soluble chitosan may be almost saturable up to 0.0032% (
w
/
v
), and sodium caprate at concentrations up to 0.64 mg/mL was safe, but water-soluble chitosan at different concentrations was all safe for these cells. In pharmacokinetics study, water-soluble chitosan at dosage of 50 mg/kg improved the bioavailability of FTA in SHL to the greatest extent, and was safe for gastrointestine from morphological observation. Besides, treatment with SHL with water-soluble chitosan at dosage of 50 mg/kg prevented MDCK damage after influenza virus propagation better significantly than that of control.
Conclusion:
Water-soluble chitosan at dosage of 50 mg/kg might be safe and effective absorption enhancer for improving the bioavailability of FTA and the antivirus activity
in vitro
in SHL.
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Compounds isolated from
Ageratum houstonianum
inhibit the activity of matrix metalloproteinases (MMP-2 and MMP-9): An oncoinformatics study
p. 18
Anupriya Verma, Syed Mohd Danish Rizvi, Sibhghatulla Shaikh, Mohd Afaque Ansari, Shazi Shakil, Fauzia Ghazal, Mohd Haris Siddiqui, Mohd Haneef, Ajijur Rehman
DOI
:10.4103/0973-1296.126653
PMID
:24695379
Background:
In osteosarcoma tissue, both MMP-2 and MMP-9 are over expressed compared to their expression in non-affected stromal tissue. Hence, gelatinases are attractive targets for anti-osteosarcoma drugs.
Objective:
To study the inhibitory activity of compounds isolated from Ageratum houstonianum against MMP-2 and MMP-9 by in-silico approach.
Material and Methods:
We performed docking study using 'Autodock 4.2' between 1,2-benzenedicarboxylic acid-bis (2-ethylhexyl) ester; squalene; 3,5-bis (1,1-dimethylethyl) phenol; pentamethyl tetrahydro-5H-chromene; (1, 4-cyclohexylphenyl) ethanone and 6-vinyl-7-methoxy-2,2-dimethylchromene with MMP-2 and MMP-9.
Results:
Among all six compounds isolated from Ageratum houstonianum, (1, 4-cyclohexylphenyl) ethanone showed the maximum potential as a putative inhibitor of both MMP-2 and MMP-9 enzymes with reference to ΔG (−7.95 and −8.2 kcal/mol, respectively) and Ki (1.48 and 0.98 μM, respectively) values. Total intermolecular energy of docking for (1, 4-cyclohexylphenyl) ethanone-MMP catalytic domain-interaction was found to be −8.55 kcal/mol for MMP-2 and −9.21 kcal/mol for MMP-9.
Conclusion:
This study explores molecular interactions between human MMPs (MMP-2 and MMP-9) and six natural compounds. This study predicts that (1,4-cyclohexylphenyl) ethanone is a more efficient inhibitor of human MMP-2 and MMP-9 enzymes compared to the other natural compounds used in this study with reference to Ki and ΔG values.
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Heartwood extract of
Acacia catechu
induces apoptosis in human breast carcinoma by altering bax/bcl-2 ratio
p. 27
Nikhil Baban Ghate, Bibhabasu Hazra, Rhitajit Sarkar, Nripendranath Mandal
DOI
:10.4103/0973-1296.126654
PMID
:24695415
Background:
The heartwood extract of
A. catechu,
called pale catechu or "Katha" in Hindi has been widely used in traditional Indian medicinal system. Although various pharmacological properties of this plant had been reported previously, only a few were concerned with the anticancer activity of this plant.
Objective:
The objective was to assess the
in vitro
anticancer and apoptosis inducing effect of 70% methanolic extract of "Katha" (ACME) on human breast adenocarcinoma cell line (MCF-7).
Materials and Methods:
MCF-7 cell line was treated with increasing concentrations of ACME and cell viability was calculated. Flow cytometric methods were used to confirm the apoptosis promoting role of ACME. Morphological changes were then analysed using confocal microscopy. Western blotting was then performed to investigate the expression of apoptogenic proteins and to analyse the activation of caspases.
Results:
ACME showed significant cytotoxicity to MCF-7 cells with an IC
50
value of 288.85 ± 25.79 μg/ml. Flow cytometric analysis and morphological studies confirmed that ACME is able to induce apoptosis in MCF-7 cells. Furthermore, immunoblot results suggested the pathway of apoptosis induction by increasing Bax/Bcl-2 ratio which results in the activation of caspase-cascade and ultimately leads to the cleavage of Poly adeno ribose polymerase (PARP).
Conclusion:
These results provide the evidence that ACME is able to inhibit the proliferation of MCF-7 cells by inducing apoptosis through intrinsic pathway.
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High-performance liquid chromatography determination and pharmacokinetics of coumarin compounds after oral administration of Samul-Tang to rats
p. 34
Youn-Hwan Hwang, Won-Kyung Cho, Doorye Jang, Jeong-Ho Ha, Jin Yeul Ma
DOI
:10.4103/0973-1296.126656
PMID
:24696544
Background:
Samul-tang has been traditionally used for the treatment of cardiovascular, gynecologic, cutaneous, and chronic inflammation disorders. Although coumarin compounds do have various pharmacological activities and the same may be present in Samul-tang, however there is little information about it.
Objective:
A simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for the determination of nodakenin, nodakenetin, decursin, decursinol, and decursinol angelate in rat plasma. To obtain a better understanding for pharmacological properties of Samul-tang, pharmacokinetic study of coumarin compounds was performed after oral administration of Samul-tang in rats.
Materials and Methods:
Chromatographic separation of the analytes was successfully achieved on a Phenomenex Luna C
18
column (4.6 mm×250 mm, 5 μm) using a mobile phase composed of acetonitrile water with a gradient elution at a flow rate of 1 mL/min. Noncompartmental analysis was performed.
Results:
Calibration curves for all analytes had good linearity (r
2
<0.999) in a wide linear range. The lower limit of quantification (LLOQ) ranged from 0.05 to 0.1 μg/mL. The variation of intra- and interday assay was less than 15%. Nodakenin, nodakenetin, and decursinol were determined in rat plasma after oral administration of Samul-tang.
Conclusion:
This developed and validated HPLC method was successfully applied to the pharmacokinetic study of three coumarin compounds in rats, given as a single oral administration of Samul-tang. These pharmacokinetic data of the nodakenin, nodakenetin, and decursinol could offer a new point of view to evaluate the pharmacological effects of Samul-tang.
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Aliphatic and eudesmalolide esters extracted from the roots of
Inula racemosa
Hook
p. 40
Maria Khan, SR Mir, Abuzar Ali, Mohammad Yusuf, Mohammed Ali
DOI
:10.4103/0973-1296.126657
PMID
:24695458
Background:
Phytochemical investigation of hydroalcoholic extract of the root of
Inula racemosa
Hook.
Materials and Methods:
Open silica gel column chromatographic techniques with different solvent systems was used for isolation of aliphatic and eudesmalolide esters from hydroalcoholic extract of the root of
Inula racemosa.
The structure elucidation of the compounds was done on the basis of spectral data analysis, chemical reactions and comparision with literature data.
Results:
Phytochemical investigation of the hydroalcoholic extract of the root of
Inula racemosa
Hook. f. led to the isolation of (5z, 13z)-n-decanyl-n-docos-5, 13-dienoate, a new fatty acid ester, two new sesquiterpenic ester identified as 15-[(13z, 18'z, 20'z)-n-tricos-13, 18, 20-trienyl]-eudesmal-4 (11), 6, 12 (13)-trien-8,14-olide-15-oate and 15- [(16'z), (21'z)-n- tetracos-16', 21'- dienyl]-eudesmal-4 (11) 6, 12 (13)-trien-8, 14-olide-15-oate, two new eudesmanolide ester i.e. 15-[(16z)-n-monadec-16'- enyl]-eudesmal-4 (11) 6, 12 (13)-trien-8,14-olide-15-oate and 15-[(16'z)-n-tetracos-16'- enyl]-endesmal-4 (11), 6, 12 (13)-trien-8,14-olide-15-oate along with the known compound n-Hexadecanyl n-docosanoate.
Conclusion:
Five new phytoconstituents were identified along with one known compound as aliphatic and eudesmalolide esters from the hydroalcoholic extract of the root of
Inula racemosa
, as mentioned above.
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Role of different types of potassium channels in the relaxation of corpus cavernosum induced by resveratrol
p. 47
Selvinaz Dalaklioglu, G Ozbey
DOI
:10.4103/0973-1296.126658
PMID
:24696545
Background:
Resveratrol (RVT), one of the most commonly employed dietary polyphenol, is used in traditional Japanese and Chinese medicine for treatment of cardiovascular diseases. Recently, we have shown that RVT has a potent relaxant effect on rat corpus cavernosum via endothelium-dependent and -independent mechanisms.
Objective:
The present study addressed the question whether different types of potassium channels are involved in the endothelium-dependent and -independent mechanism of corpus cavernosum relaxation induced by RVT.
Materials
and
Methods:
Strips of corpus cavernosum from rats were mounted in an organ-bath system for isometric tension studies.
Results:
RVT (1-100 μmol/L) produced concentration-dependent relaxation responses in rat corpus cavernosum pre-contracted by phenylephrine. The non-selective potassium channels blocker tetraethylammonium chloride (TEA, 10 mmol/L), ATP-sensitive potassium (K
ATP
) channels blocker glibenclamide (10 μmol/L), and inward rectifier potassium (Kir) channels inhibitor barium chloride (BaCl
2
, 30 μmol/L) caused a significant inhibition on the relaxation response to RVT, whereas voltage-dependent potassium channels inhibitor 4-aminopyridine (4-AP, 1 mmol/L), and large conductance calcium-activated potassium (BK
Ca
) channels inhibitor iberiotoxin (IbTX, 0.1 μmol/L) did not significantly alter relaxant responses of corpus cavernosum strips to RVT. In addition, relaxant responses to RVT did not significantly inhibited by the combination of selective inhibitors of small and intermediate conductance BK
Ca
channels (0.1 μmol/L charybdotoxin and 1 μmol/L apamin, respectively).
Conclusion:
These results demonstrated that endothelial small and intermediate conductance BK
Ca
channels are not thought to be an important role in RVT-induced endothelium-dependent relaxation of corpus cavernosum. The endothelium-independent corpus cavernosum relaxation induced by RVT is seems to largely depend on Kir channels and K
ATP
channels in corporal tissue.
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Fatty acid composition and preclinical resarches on
Anthemis wiedemanniana
Fisch. & Mey.: Discovery of a new anti-inflammatory agent
p. 53
Tuba Mert Gönenç, Esra Küpeli Akkol, Ipek Süntar, Tuğçe Fafal Erdoğan, Bijen Kıvçak
DOI
:10.4103/0973-1296.126660
PMID
:24696546
Background:
Anthemis
species have been used for the treatment of gastrointestinal disorders, hemorrhoid, stomachache and inflammatory diseases in Turkish folk medicine.
Anthemis wiedemanniana
Fisch. And Mey. is an endemic plant used as painkiller, antispasmodic, sedative and for the treatment of urinary inflammations.
Objective:
The objective of the present study is to evaluate the anti-inflamatory activity of the extracts of
A. wiedemanniana
by using
in vivo
methods.
Materials and Methods:
Carrageenan-, PGE
2
- and serotonin-induced hind paw edema, 12-
O
-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema and acetic acid-induced increase in capillary permeability models were used for the anti-inflammatory activity assessment. Moreover, the fatty acid composition of
A. wiedemanniana
was investigated by gas chromatography (GC).
Results:
n
-Hexane, diethyl ether and total sesquiterpene lactone extracts exhibited significant inhibition in carrageenan-induced hind paw edema and acetic acid-induced increase in capillary permeability model.
n
-Hexane and total sesquiterperne lactone extracts showed anti-inflammatory activity in PGE
2
- and serotonin-induced hind paw edema model. However none of the extracts displayed significant activity in TPA-induced ear edema model in mice. C
4:0
(Buthyric acid), C
20:0
(Arachidic acid) and C
16:1
(Palmitoleik acid) were found to be the major fatty acids in these species. Saturated fatty acids (SFA) were found in higher amounts than monounsaturated fatty acids and polyunsaturated fatty acids. SFAs were determined as 63.17%, UFAs as 20.89% and PuFAs as 15.95%.
Conclusion:
This study confirms the traditional usage of
A. wiedemanniana
for inflammatory diseases.
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Curcumin attenuates neurotoxicity induced by fluoride: An
in vivo
evidence
p. 61
Chhavi Sharma, Pooja Suhalka, Piyu Sukhwal, Neha Jaiswal, Maheep Bhatnagar
DOI
:10.4103/0973-1296.126663
PMID
:24696547
Background:
Curcumin (Cur), an active ingredient of turmeric is known to have multiple activities, including an antioxidant property and has been suggested to be useful in treatment of several neurological diseases.
Objective:
To investigate the neuroprotective effects of Cur to mitigate the effect of the Fluoride (F) induced neurotoxicity in mice brain using the histological and the biochemical parameters.
Materials
and
Methods:
Exposure of mice (30 days old male) to F (120 ppm) daily for 30 days.
Result
and
Discussion:
Treatment with the F causes an increase in lipid peroxidation (LPO) and also increase in the neurodegenerative cells in the hippocampal sub-regions. Interestingly, co-treatment with Cur (30 mg/kg BW) with F (120 ppm) for 30 days results in significant decreases in LPO with a concomitant decrease in neurodegeneration as compared with those treated with F alone.
Conclusion:
Our study reveals that Cur is useful in ameliorating degenerative effects of F in mice brain.
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Fraction and chemical analysis of antioxidant active polysaccharide isolated from flue-cured tobacco leaves
p. 66
Chunping Xu, Chenchen Yang, Duobin Mao
DOI
:10.4103/0973-1296.126664
PMID
:24696548
Background:
The metabolic compounds from tobacco were investigated to posses various biological activities, such as antioxidant, antimicrobial and neuroprotective activities.
Materials and Methods:
Polysaccharides were isolated from the ultrasound-assisted extraction of flue-cured tobacco with hot water. The purified polysaccharides were analyzed by GC-MS, FT-IR, and TGA (Thermogravimetric analysis). The antioxidant activities of purified polysaccharides were evaluated in vitro.
Results:
Two polysaccharides (Fr-I and Fr-II) were isolated and purified. The analysis of monosaccharide composition in the polysaccharide by GC/MS revealed that Fr-I and Fr-II were heteropolysaccharides. Fr-I consisted of D-mannose and galactose, ribose and arabinose, while Fr-II was mainly composed of glucose, allose, galactose and mannose. Thermo gravimetric analysis (TGA) indicated that the degradation temperature (Td) of the Fr-I (241 °C) was higher than that of Fr-II (220 °C). Both samples showed inhibitory effects on OH (hydroxyl) and DPPH (2, 2-diphenyl-1-picryl-hydrazyl-hydrate) radical in a concentration-dependent manner. Comparing Fr-I with Fr-II, the latter has a strong scavenging ability.
Conclusion:
Both polysaccharide fractions showed significant antioxidant effects. Various factors influenced the antioxidant activity of polysaccharides.
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Cytotoxic activity of
Alpinia murdochii
Ridl.: A mountain ginger species from Peninsular Malaysia
p. 70
Kae Shin Sim, Halijah Ibrahim, Sri Nurestri Abdul Malek, Devi Rosmy Syamsir, Khalijah Awang
DOI
:10.4103/0973-1296.126666
PMID
:24695515
Background:
Alpinia
murdochii
(Zingiberaceae) is a wild ginger species restricted to mountain areas of Peninsular Malaysia. Due to rapid development and deforestation activities, this species is becoming rare. This is the first report of the cytotoxic activity of
A.
murdochii
.
Objective:
The present study aimed to investigate the cytotoxic effect of leaves and rhizomes of
A.
murdochii
against selected human cancer cell lines by using
in vitro
cytotoxicity assay.
Materials and Methods:
The leaves and rhizomes of
A.
murdochii
were extracted in hexane, dichloromethane (CH
2
Cl
2
), and methanol (MeOH) prior to cytotoxic activity assessment against selected human cancer cell lines, namely MCF7 (hormone dependent breast carcinoma cell line), HT29 (colon carcinoma cell line), and SKOV-3 (ovarian cancer cell line) by using
in vitro
neutral red cytotoxicity assay.
Results:
The hexane and CH
2
Cl
2
extracts of both leaves and rhizomes exhibited remarkable cytotoxic effect against SKOV-3 cells with the IC
50
values in the range of 5.2-16.7 μg/ml.
Conclusion:
Based on the preliminary data obtained in the present study, the leaves and rhizomes of
A.
murdochii
may be viable therapeutic or preventive candidates for the treatment of ovarian cancer.
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Acridone alkaloids with cytotoxic and antimalarial activities from
Zanthoxylum simullans
Hance
p. 73
Chao Wang, Jinfu Wan, Zhinan Mei, Xinzhou Yang
DOI
:10.4103/0973-1296.126669
PMID
:24696549
Background:
Zanthoxylum
simullans
Hance is a popular natural spice belonging to the Rutaceae family and it is one of the common prescribed herbs in traditional Chinese medicine.
Materials
and
Methods:
The chemical constituents were mainly isolated and purified by silica gel column chromatography and semi-preparative High Performance Liquid Chromatography. Their structures were identified by comparing the spectral data with those reported in the literature. Cytotoxic activities for the isolated acridone alkaloids were evaluated against two prostate cancer cell lines PC-3M and Lymph Node Carcinoma of Prostate (LNCaP), and their antimalarial activities were tested against two different strains of the parasite
Plasmodium
falciparum
3D7, and Dd2.
Results:
The root bark MeOH extract of
Z.
simullans
Hance afforded β-sitosterol, 4-methoxy benzoic acid, daucosterol, and five acridone alkaloids, normelicopidine, normelicopine, melicopine, melicopidine, and melicopicine. All five acridone alkaloids were isolated from this plant for the first time and exhibited certain cytotoxic and antimalarial activities
in
vitro
.
Conclusion:
Normelicopidine was the most active against PC-3M, LNCaP and Dd2 with IC
50
values of 12.5, 21.1, and 18.9 ug/mL respectively.
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Optimal inductive and cultural conditions of
Polygonum multiflorum
transgenic hairy roots mediated with
Agrobacterium rhizogenes
R1601 and an analysis of their anthraquinone constituents
p. 77
Bing Huang, Huanjie Lin, Chuanyan Yan, Hongyan Qiu, Lipeng Qiu, Rongmin Yu
DOI
:10.4103/0973-1296.126671
PMID
:24696550
Background:
Polygonum multiflorum
is an important medicinal plant. Hairy roots systems obtained by transforming plant tissues with the natural genetic engineer
Agrobacterium rhizogenes
can produce valuable biological active substances, which have immense potential in the pharmaceutical industry.
Objective:
To optimize the inductive and cultural conditions of
P. multiflorum
hairy roots and to identify the major active secondary metabolites in hairy roots.
Materials and Methods:
P. multiflorum
hairy root were mediated with
A. rhizogenes
R1601 to induce hairy roots. Four combinations, including Murashige-Skoog (MS), 1/2 MS, B
5
, and White, were investigated to optimize the culture medium. MS medium was selected for the growth measurement. The qualitative and quantitative determinations of free anthraquinone in hairy roots were compared with the calli and aseptic plantlets using high-performance liquid chromatography.
Results:
The inductive rates of hairy roots by leaves were higher than for any other explants. The presence of agropine in the
P. multiflorum
hairy roots confirmed that they were indeed transgenic. MS medium was the most suitable of the four media for hairy root growth. Meanwhile, the growth kinetics and nutrient consumption results showed that the hairy roots displayed a sigmoidal growth curve and that their optimal inoculation time was 18-21 days. The determination of the anthraquinone constituents indicated that the rhein content of the hairy roots reached 2.495 μg g
−1
and was 2.55-fold higher than that of natural plants.
Conclusion:
Transgenic hairy roots of
P. multiflorum
could be one of the most potent materials for industrial-scale production of bioactive anthraquinone constituents.
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A validated high performance liquid chromatograph-photodiode array method for simultaneous determination of 10 bioactive components in compound hongdoushan capsule
p. 83
Liancai Zhu, Xian Yang, Jun Tan, Bochu Wang, Xue Zhang
DOI
:10.4103/0973-1296.126673
PMID
:24696551
Background:
The compound Hongdoushan capsule (CHC) is widely known as compound herbal preparation and is often used to treat ovarian cancer and breast cancer, and to enhance the body immunity, etc., in clinical practice.
Objective:
To determine simultaneously 10 bioactive components from CHC, namely glycyrrhetinic acid, liquiritin, glycyrrhizin, baccatin III, 10-deacetylbaccatin III, cephalomannine, taxol, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rb1.
Materials and Methods:
A high performance liquid chromatograph method coupled with photodiode array detector was developed and validated for the 1
st
time. Chromatographic analysis was performed on a SHIMADZU C
18
by utilizing a gradient elution program. The mobile phase was acetonitrile (A)-water (B) at a flow rate of 0.8 mL/min.
Results:
The calibration curve was linear over the investigated concentration ranges with the values of
r
2
higher than 0.9993 for all the 10 bioactive components. The average recovery rates range from 98.4% to 100.5% with relative standard deviations ≤2.9%. The developed method was successfully applied to analyze 10 compounds in six CHC samples from different batches. In addition, the herbal sources of 32 chromatographic peaks were identified through comparative studying on chromatograms of standard, the respective extracts of Hongdoushan, RenShen, GanCao, and CHC.
Conclusion:
All the results imply that the accurate and reproducible method developed has high separation rate and enables the determination of 10 bioactive components in a single run for the quality control of CHC.
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