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Objective To study the mechanism of Puerariae Radix in the treatment of ischemic stroke by using molecular docking technology. Methods The small molecules of Puerariae Radix based on molecular docking technology docked with 21 key targets protein of cerebral ischemic stroke, and multi-component protein target network was established by Cytoscape 3.1.1 software. Results Through virtual screening of molecular docking technology, 28 active small molecules of Puerariae Radix were chosen, 12 of which were novel small molecules, and it identified that 11 of those compounds had strong interactions with no less than 10 targets. Conclusion The molecular docking can be used to find the active components of Puerariae Radix in treatment of cerebral ischemic stroke, which provide a new reference of studying on the multiple ingredients and targets of Chinese materia medica compounds.
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Objective: To identify the flavonoids in Gegen Zhiju Soft Capsule (a soft capsule prepared with Puerariae Radix and Hoveniae Dulcis Fructus seu Semen and so on) using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Methods: The analysis was performed on Agilent Poroshell 120 SB-C18 column (100 mm ×2.1 mm, 2.7 μm) gradient elution with a mobile phase of 0.1% formic acid aqueous solution and acetonitrile. The flow rate was at 0.4 mL/min. TOFMS was applied for qualitative analysis, and the data were collected by the negative ion mode using ESI ion source. The parameters of ion source were as follows: drying gas temperature, 300 ℃; drying gas flow rate, 10 L/min; nebulizer gas pressure, 276 kPa, sheath gas temperature, 350 ℃; sheath gas flow rate, 11 L/min; capillary voltage, 3 500 V; fragmentor voltage, 175 V. Results: Twenty flavonoid compounds were identified by TOF-MS and literature data. Relative molecular weight was concentrated within 250-650.Conclusion: The rapid separation with UPLC and Q-TOF-MS determination of the precise molecular weight information could effectively analyze and identify the flavonoid compounds in Gegen Zhiju Soft Capsule. The method is accurate, rapid, and sensitive, and could provide a reliable and effective technique for the quality control.
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Objective: To study the powder characteristics and dissolution behavior of the composition in decoction for boiling powders of Chinese materia medica, and to provide scientific evidence for particle size control and its application. Methods: Four kinds of herbal pieces such as Scutellariae Radix, Coptidis Rhizoma, Puerariae Radix, and Glycyrrhizae Preparata Radix were prepared in powders. Powder characteristics including particles distribution and mobility, and dispersion properties of particles in the water decoction were investigated. Additionally, dissolution behaviors of the composition in decoction made from powders and pieces were compared. Results: Well-distributed particles and high mobility were observed in Scutellariae Radix, Coptidis Rhizoma, and Glycyrrhizae Radix Preparata powders, rather than in Puerariae Radix powder. The suspended particles in the decoction in both powders and pieces were submicron. However, optical and electrical properties showed that the suspended particles in the decoction made from powders were more, bigger, and easier to settle as heterogeneous liquid was. Dissolution velocity and amount of the active ingredients in the decoction made from powders were significantly higher than those of pieces. Conclusion: As a new type of herbal decoction which could increase the dissolution of active ingredients, it could shorten decocting duration and improve the availability of herbs. The boiling powders are worth being studied and promoted.
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Objective: To prepare Puerariae Radix Flavones Dropping Pills and to investigate the dissolution. Methods: Exterior quality, weight variation, and resolving time were used as comprehendsive evaluation indicators to select dropping conditions by orthogonal design. HPLC was used to determine the content of puerarin and rotating basket method was used to determine the dissolution rate in vitro of the dropping pills and tablets. Results: The optimal preparation conditions for the pills were as follows: proportion of drug and matrix was 1:3, drop rate was 20 d/min, the temperature of drug fluids was 80°C, the condensate tube outlet temperature was 50°C, dimethylsilicone was the refrigerant at the temperature of 10°C and 6 cm distance above liquid level. The content of peurarin in the dropping pills was 5.542 mg/g. The accumulated dissolution rate of puerarin in the dropping pills reached 98.81% in 20 min, while the accumulated dissolution rate of puerarin in tablets was only 10.70% in 20 min. Conclusion: The preparation process and HPLC determination method are simple, stable, and feasible. The dissolution rate of puerarin can be improved in the Dropping pills.
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Objective To investigate the effects on proliferation of multiple myeloma cell lines U266 and RPMI 8226 induced by puerariae radix flavones (PRF) in vitro and its possible mechanism.Methods Exposed to 0,10,30,50,100 μg/ml PRF for 48 h and 72 h,the U266 and RPMI 8226 cells proliferation inhibitory rates were detected by MTT assay,cell cycles by flow cytometry (FCM),morphologic changes of U266 cells by Wright' s staining,and early-stage apoptotic rates of U266 cells by FITC-Annexin V/PI staining with FCM.Analysis of DNA fragment was made to test characteristic apoptosis DNA ladder in U266 cells.Results 0,10,30,50,100 μg/ml PRF could inhibit the proliferation of U266 and RPMI 8226 cells in a dose-dependent manner (U266 ＞ RPMI 8226).Cell cycle analyses in U266 and RPMI 8226 cells showed that sub-diploid peaks,but cell cycles changed minor.Wright's staining of U266 cells showed hardly any apoptostic character istic.Annexin V/PI double staining indicated that early-stage apoptotic rates of U266 cells exposed to 0,10,30,50,100 μg/ml PRF for 48 h were mildly increased in a dose-dependent manner.They were (3.20±0.36) ％,(5.20±0.92) ％,(7.30±1.22) ％,(8.10±0.53) ％ and (10.80±0.90) ％,respectively.The group differences had statistical significance (P ＜ 0.05).Analysis of DNA fragment barely exhibited the characteristic DNA ladder in U266 cells.Conclusion A certain concentrations of PRF could inhibit the proliferation of U266 and RPMI 8226 cells significantly.It is suggested that apoptosis related to the proliferative inhibition mechanism induced by PRF in U266 cell line,but not main.Other pathways such as necrosis and autophagy whether or not involved need further investigation.
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This study was performed to investigate the effect of Puerariae radix-ethanol extracts rich in isoflavone on the antioxidative system of rats. For this purpose, first, Puerariae radix was extracted with ethanol, and its total isoflavone and puerarin contents were analysed. Second, female Sprague Dawley rats were fed for 6 weeks with four diets which were based on AIN96G diet and supplemented with Puerariae radix-ethanol extracts to contain isoflavone. The isoflavone contents of four experimental diets were 0 mg, 500 mg, 1,000 mg, 2,000 mg per kg diet, respectively (control, P0.05%, P0.1%, P0.2%). Liver and erythrocyte activities of antioxidative enzyme such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GSHpx) were measured. Also, plasma and liver malondialdehyde (MDA) concentrations, liver glutathione (GSH) and oxidized glutathione (GSSG) concentrations were measured. The total isoflavone content of Puerariae radix-ethanol extract was 3067.6 mg per 100 g extract and the content of puerarin was 2557.4 mg per 100 g extract. The erythrocyte activities of GSH-Px and catalase were higher in group P0.1% and P0.2%. But SOD activity of erythocyte did not show any difference by the Puerariae radix-ethanol extract supplementation in diet. The activity of SOD in liver increased significantly by the supplementation of extract, showing highest level in P0.1% group. The liver GSH concentration increased significantly in group of P0.05%, P0.1%, and P0.2% compared with control group (p < 0.05). The GSSG concentration in liver showed no difference by the supplementation of Puerariae radix extract from the control group, except P0.2% group. The plasma MDA concentration did not show any significant differences by the extract supplementation. But the liver MDA concentration decreased by the extract supplementation, showing the lowest level in P0.1% diet group. These results suggest that the supplementation of Puerariae radix-ethanol extract can inhibit lipid peroxidation in liver and enhance the antioxidative defense competence of rats.
Subject(s)
Animals , Female , Humans , Rats , Catalase , Diet , Erythrocytes , Ethanol , Glutathione , Glutathione Disulfide , Glutathione Peroxidase , Lipid Peroxidation , Liver , Malondialdehyde , Mental Competency , Plasma , Pueraria , Rats, Sprague-Dawley , Superoxide DismutaseABSTRACT
In the previous paper, the authors have investigated a Chinese crude drug Ge-gen, the root of Pueraria species of the family Leguminosae, and concluded that it should be collected in the early summer for medical use. This is a report of the differences in the content of the constituents between Ge-gen harvested in the summer and in the winter. Yields of water extract of winter Ge-gens were generally higher, while no significant difference was recognized in the content of starch and flavonoids, such as puerarin and daidzin. Starch content varies widely with the stock, and that of the same stock did not show significant difference between the summer root and the winter one.
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Ge-gen, a Chinese crude drug derived from the root of <i>Pueraria lobata</i> Ohwi of the family Leguminosae, is collected in the winter nowadays both in China and in Japan. However, in the old descriptions of materia medica written in ancient China, it was said that the proper collection season was May or in the early summer on Chinese calendar. Present herbological study resulted that the collection season had been changed in the Yuan or Ming dynasty in China, mainly because of the confusion with eating Ge-gen, which was collected in the winter. The Ge-gen collected in the early summer should be dispensed in the prescriptions written in the old medical books such as Shang-han-lun and Jin-kui-yao-lue.