ABSTRACT
Objective: To investigate the effects of five kinds of flavonoids (calycosin, formononetin, ononin, isoliquiritigenin, and medicarpin) from Hedysari Radix on promoting osteogenic differentiation of rat bone marrow stromal cells (rBMSCs) and rat calvarial osteoblasts (ROBs). Methods: rBMSCs were isolated according to plastic adherence. ROBs were isolated by enzyme digestion method. The proliferation of rBMSCs and ROBs were detected by MTT assay. ALP activity and calcium content of rBMSCs and ROBs cells were detected by alkaline phosphatase kit and calcium kit. Mineralized nodule formation was detected by alizarin red staining. Results: The five components could promote proliferation, increase ALP activity, increase calcium content, and increase the area and number of calcified nodules of rBMSCs and ROBs (P < 0.05). Among them, calycosin had the best effect on promoting the osteogenic differentiation of rBMSCs, and medicarpin promoted the osteogenic differentiation of ROBs with the best effect, followed by calycosin. Conclusion: Five flavonoids promoted the improvement of osteogenic function, while calycosin has better osteogenic activity on rBMSCs and ROBs and can be used as an excellent osteoinductive factor.
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Objective: To study the correlation of HPLC fingerprint of Hedysari Radix and efficacy of improving immunity of normal mice, and provide the basis for finding bioactive components and selecting the index composition of quality control. Methods: The mice delayed type hypersensitivity (DTH) model induced by dinitoflruorobenzene (DNFB) was used to study the effect and then to perform ig administration with different extracts of Hedysari Radix for selecting the best part. To study the correlation between fingerprint and efficacy and find out the effective substances foundation of improving immunity, the peak area of each common peak from HPLC fingerprint was associated with the date of improving immunity in the method of partial least square regression (PLS). Results: In ear swelling experiment, water extract of Hedysari Radix was the most effective part which could improve the immunity function. PLS was used to evaluate the correlation of effect of improving immunity and components of the water extract including adenosine, ononin, genistein, formononeti, and medicarpin, the index of compounds to characterize the quality of Hedysari Radix, showed the negative correlation with the effectiveness. Conclusion: A new method is established to evaluate the activity of enhancing immunity of Hedysari Radix. It is of practical significance as an effective approach for controlling quality and exploring the material basis for efficacy of traditional Chinese medicine compounds.
ABSTRACT
Eight isoflavonoid compounds were isolated from the EtOAc fraction of Maackia amurensis which had shown the highest anti-Helicobacter pylori activity among the fractions, using medium pressure liquid chromatography and recrystallization. Based on the spectroscopic data including 1H-NMR, 13C-NMR, HMBC and MS data, the chemical structures of the isolates were determined to be (-)-medicarpin (1), afromosin (2), formononetin (3), tectorigenin (4), prunetin (5), wistin (6), tectoridin (7) and ononin (8). Anti-H. pylori activity of each compound was evaluated with broth dilution assay. As a result, (-)-medicarpin (1), tectorigenin (4) and wistin (6) showed anti-H. pylori activity. (-)-Medicarpin (1) exhibited the most potent growth inhibitory activity against H. pylori with the minimal inhibitory concentration (MIC)90 of 25 microM, and tectorigenin (4) with MIC90 of 100 microM ranked the second. This is the first study to show the anti-H. pylori activity of M. amurensis, and it is suggested that the stem bark of M. amurensis or the EtOAc fraction or the isolated compounds can be a new natural source for the treatment of H. pylori infection.
Subject(s)
Chromatography, Liquid , MaackiaABSTRACT
Objective:To establish an HPLC method for the determination of calycosin-7-glucoside, genistein, formononetin and medicarpin in Radix Hedysari. Methods: The sample was refluxed with methanol in a water bath. The HPLC was performed on an SunFire C18 column(250 mm × 4. 6 mm,5 μm) with acetonitrile-0. 2% H3 PO4 as the mobile phase by gradient elution. The flow rate was 1. 0 ml·min-1 and the column temperature was at 35℃. The detection wavelength was 240 nm and the sample size was 20 μl. Results:The linear range of calycosin-7-glucoside was 0.035-1.042 μg(r=0.999 6), that of genistein was 0.027-0.821 μg(r=0. 999 7), that of formononetin was 0. 031-0. 941 μg(r=0. 999 9) and that of medicarpin was 0. 025-0. 745 μg(r=0. 999 6). The average recovery of calycosin-7-glucoside, genistein, formononetin and medicarpin was 100. 32%(RSD=1. 87%), 99. 3%(RSD=1. 76%), 100. 5%(RSD=1. 48%) and 99. 2%(RSD=1. 45%)(n=6), respectively. Conclusion:The method shows short analyt-ic time, good stability and promising operation accuracy, which provides the reference for the quality control of Radix Hedysari.