ABSTRACT
objective To explore the role of Xanthatin in the targets of epithelial-mesenchymal transition (EMT) process using molecular docking method,and the effect on the target protein expression of HepG2 cells was detected by Western assay.Method Dhs,Vimentin,Snail and VEGFR3 are critical targets in EMT process,the spatial binding ability of Xanthium was evaluated by molecular docking method,compared with the corresponding endogenous substances:nicotinarnide adenine dinucleotide,Acetate ion,flavin adenine dinucleotide,and N-Acetyl-D-glucosamine.HepG2 cells were cultured,and the effects of Xanthatin of 1,5 and 20 mol/L concentrations on Dhs,Vimentin,Snail and VEGFR3 protein expression were detected by Western Blotting assay.Rusult Molecular docking show that Xanthatin has obvious affinity to key factors of EMT process such as Dhs,Vimentin,and VEGF-R3,higher than that of endogenous substance;and the affinity with Vimentin was less than that of endogenous substance;Western Blotting experiments proved the virtual results.The expression of Vimentin,Snail,VEGFR3 protein was significantly lowered,and the expression of e-cadherin was significantly raised.Conclusion The influence of Xanthatin to key factor e-cadherin,Vimentin,Snail,VEGFR3 are obvious,Which is likely to be a potential target.The results of computer virtual experiment and Western Blotting have certain similarity.Molecular virtual docking can pre hint the potential target factor.
ABSTRACT
Objective To establish a method for determining the contents of polyphyllin Ⅰ and polyphyllin Ⅱ in Kangkeling Mixture. Methods The contents of polyphyllin Ⅰ and Ⅱ were determined by HPLC gradient elution. Poroshell 120 Ec-C18 column (4.6 mm × 150 mm, 4 μm) was used; Acetonitrile-water (A:B) was set as the mobile phase; the flow rate was 1.0 mL/min; the detection wavelength was 210 nm; column temperature was 30 ℃. Results Polyphyllin Ⅰ showed good linear relationship in the range of 1.009–10.09 μg (r=0.999 6), and the average recovery was 97.31% (RSD=2.05%, n=6). Polyphyllin Ⅱ showed good linear relationship in the range of 0.640 5–6.405 μg (r=0.999 8), and the average recovery was 96.41% (RSD=1.67%, n=6). Conclusion The method is simple, with good repeatability and accurate results, which can be used to determine the contents of polyphyllin Ⅰ and Ⅱ in Kangkeling Mixture.
ABSTRACT
Objective To establish a method for determining the contents of polyphyllin Ⅰ and polyphyllin Ⅱ in Kangkeling Mixture. Methods The contents of polyphyllin Ⅰ and Ⅱ were determined by HPLC gradient elution. Poroshell 120 Ec-C18 column (4.6 mm × 150 mm, 4 μm) was used; Acetonitrile-water (A:B) was set as the mobile phase; the flow rate was 1.0 mL/min; the detection wavelength was 210 nm; column temperature was 30 ℃. Results Polyphyllin Ⅰ showed good linear relationship in the range of 1.009–10.09 μg (r=0.999 6), and the average recovery was 97.31% (RSD=2.05%, n=6). Polyphyllin Ⅱ showed good linear relationship in the range of 0.640 5–6.405 μg (r=0.999 8), and the average recovery was 96.41% (RSD=1.67%, n=6). Conclusion The method is simple, with good repeatability and accurate results, which can be used to determine the contents of polyphyllin Ⅰ and Ⅱ in Kangkeling Mixture.