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Objective To study the correlation of anti-osteoporosis effect of Hedysarum polybotrys and HPLC fingerprint. Methods Osteoporosis model was established through ovary resection of female rats to select the best part after performing ig administration with different extracts of H. polybotrys. The fingerprints of ethanol extract of ten batches of H. polybotrys from different habitats were established by HPLC. The partial least squares regression (PLSR) analysis was used to explore the correlation between peak areas of HPLC fingerprints and efficacy. Moreover, the effects of active ingredient of H. polybotrys on the function of osteoblasts were observed. Results The ethanol extract of H. polybotrys was the most effective part. Nine common peaks of the active fraction were selected. Adenosine, calycosin and ononin in ethanol extract of H. polybotrys have been identified separately, among which adenosine and calycosin were positively correlated with the pharmacodynamic data, and ononin was negatively correlated with the effect of drug. Additionally, calycosin can increase osteoblast ALP activity. Conclusion The results indicate that anti-osteoporosis effect of H. polybotrys is related to various components, among which adenosine and calycosin have a certain contribution to this effect, and calycosin can also stimulate the differentiation of osteoblasts in vitro.
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Objective To study the determination method of total flavonoids in Gansu Astragali Radix and Hedysarum Polybotrys. Methods Calycosin glycosides etc. was selected as reference substances, comparison on the difference of absorption curves was done by ultraviolet spectroscopy and colorimetric method (NaNO2-Al(NO3)3-NaOH, AlCl3, Mg(Ac)2, NaOH, phosphomolybdic acid, HCl-Mg power). Results With colorimetric method, the maximum absorption wavelength of referrence and the test was inconsistent. The absorption peak shape was also different. With UV method, Calycosin glycosides in band Ⅱ (260 nm) showed a shoulder absorption. Astragali Radix and Hedysarum Polybotrys also showed characteristic shoulder absorptions in band Ⅱ with absorption wavelength at 263 nm and 265 nm. So the sample absorption wavelength is basically the same as that of the control sample. Conclusion Colorimetries usually used for determination of total flavonoids are not suitable for the comparison determination of Gansu Astragali Radix and Hedysarum Polybotrys. It is suitable for determining the contents of total flavonoids in samples by UV spectrophotometry at the band Ⅱ, which is the characteristic absorption band of isoflavone compound.
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ABSTRCT OBJECTIVE:To analyze the correlation between contents of active ingredients in Hedysarum polybotrys and ecologi-cal factors. METHODS:HPLC was used for the simultaneous determination of calycosin and formononetin in H. polybotrys;UV spectrophotometry was used to determine the content of hedysari polysaccharide;and hot-dip method was used to determine the con-tent of extracts in H. polybotrys. Factor analysis method was used to analyze the correlation between contents of active ingredients and ecological factors. RESULTS:4 common factors,which were heat,geography,illumination and precipitation,were extracted. The heat was an ecological dominant factor influencing the quality of H. polybotrys. And ≥10 ℃ accurnulated temperature was the most significant variate influencing hedysari polysaccharide,latitude was the most significant variate influencing the contents of ca-lycosin and formononetin H. polybotrys;and annual averge amount of radiation was the most significant variate influencing content of extracts in H. polybotrys. CONCLUSIONS:Different ecological factors have different effects on the contents of active ingredi-ents in H. polybotrys,and it is the result of the combined effect of various of ecological factors.
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Objective: To investigate the synergy effect of Hedysarum polysaccharides 1 (HPS1) on apoptosis of lung adenocarcinoma A549 cells induced by cisplatin, and to explore its underlying mechanism. Methods: The inhibitory effects of different concentrations of HPS1 alone and cisplatin alone and in combination on the growth of A549 cells were tested by MTT method. The apoptosis and cell cycle were detected by flow cytometry. The expressions of Bcl-2 and Bax mRNAs were measured by real-time fluorescence quantitative-PCR. Results: Different concentrations of HPS1 had inhibitory effects on the growth of A549 cells in a concentration- and time-dependent manner. The apoptotic rate was significantly increased in A549 cells treated with HPS1 in combination with cisplatin, compared with that in cisplatin-treated A549 cells (P < 0.05), and the cell cycle was arrested at phase G1+S. The expression level of Bcl-2 protein was decreased (P < 0.05) and the expression level of Bax was increased (P < 0.05) in a concentration-dependent manner in A549 cells treated with different concentrations of HPS1, and these effects were significantly enhanced in A549 cells treated with HPS1 in combination with cisplatin as compared with those in the cells treated with cisplatin alone (P < 0.05). Conclusion: HPS1 can enhance the apoptotic level of A549 cells induced by cisplatin, and this effect may be associated with the decrease in the ratio of Bcl-2/Bax mRNAs. Copyright © 2013 by TUMOR.
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Objective To study the effect of the ultra-filtration extract from Angelica sinensis and Hedysarum polybotrys’s mixture on the VEGF mRNA expression of ECV-304 cell. Method Angelica sinensis and Hedysarum polybotrys’s mixture were refined by ultra-filtration technique. ECV-304 cells were cultured as model, and its proliferation was detected by MTT colorimetry. VEGF mRNA expression was observed by semi-quantitative RT-PCR. Results The ultra-filtration extract from Angelica sinensis and Hedysarum polybotrys’s mixture could markly promote the growth of ECV-304 cells, there was significant difference between experimental and control group (P
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Objective To investigate whether the administration of the ultra-filtration extract mixture from Angelica sinensis and Hedysarum polybotrys is able to protect cardiomyocytes from oxidative injury of rats induced by H2O2 and its potential mechanism. Methods Myocardial cells from 2—3 d neonatal rats were cultured in DF medium and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from A. sinensis and H. polybotrys was given in three doses of 3.75,7.5,and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using MTT. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase (LDH) and creatine kinase (CK) releases in the medium,superoxide dismutase (SOD) activities and intracellular malondialdehyde (MDA),and myeloperoxidase (MPO) contents. The levels of caspase-3 and hsp70 expression in cardiomyocytes were measured by RT-PCR. Results The ultra-filtration extract mixture could protect the cardiomyocytes from H2O2 injury in a dose-dependent manner (3.75,7.5,and 15 mg/mL). The ultra-filtration extract mixture could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents,increase SOD activity,upregulate hsp70 expression,and downregulate caspase-3 expression. Conclusion The ultra-filtration extract mixture has protection on cardiomyocytes injured by H2O2 through improving cell antioxidant ability,upregulating hsp70 expression,and inhibiting caspase-3 activity.
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Objective To investigate the chemcial constituents in Hedysarum polybotrys and their antitumor activities.Methods The chemical constituents were isolated by various column chromatographic methods,and their structures were elucidated mainly by NMR and MS evidences;Cytotoxicities of the purified compounds against human cancer cell lines HepG2 were evaluated by MTT method.Results Eleven compounds were isolated and identified as: n-tetracosanoic acid(Ⅰ),n-hexacosanic acid(Ⅱ),trioleic glyceride(Ⅲ),glycerol monopalmitate(Ⅳ),cetyl ferulate(Ⅴ),(+)syringaresinol(Ⅵ),7-hydroxy-4′-methoxyisoflavane(Ⅶ),isoliquiritigenin(Ⅷ),3-hydroxy-9-methoxypterocarpan(Ⅸ), ?-sitosterol(Ⅹ),and daucosterol(Ⅺ).Conclusion Compounds Ⅰ—Ⅳ and Ⅵ are isolated from this plant for the first time.Compound Ⅸ shows the inhibitory activity on HepG2 with IC50 values of 10.69 ?mol/L.