The Proliferation Effect Of The Water Extraction Of Lithocarpus Polystachyrus Rehd On Normal Human Liver Cells L-02

Objective: The present study was designed to investigate the effective constituents of ST such as tea polyphenols,phlorizin and trilobatin,and establish an appropriate method for the determination of phlorizin and trilobatin of ST;Furthermore,the proliferation effects of ST and its underlying mechanisms in L-02 cells were explored.Methods: Lithocarpus polystachyrus Rehd was extract using water and it was producted to be freeze-dried powder by freenze drying.The content of tea polyphenols and phlorizin,trilobatin were detected by UV and HPLC,respectively.MTT assay was used to detect the safety range of ST;L-02 cells were divided into four groups: control group,low-concentration group(50 mg/ml),middle-concentration group(200 m g/ml),high-concentration group(800 m g/ml);Brd U ELISA and EDU staining were used to observe DNA concent and cell proliferation;Moreover,flow cytometry was applied to analyze the distribution of cell cycle.Furthermore,the expression of cyclin D1,CDK4,HGF/c-Met,Akt,Erk1/2 were detected by western blot.Result: The content of tea polyphenols,phlorizin,trilobatin were 27.65%,0.65%,18.54%,respectively.The results showed that the safety range of ST is 25mg/ml~1500 mg/ml,and ST significantly promoted L-02 cells proliferation(P<0.01)in a concentration-dependent manner,as well as increased the percentage of S phase(P<0.01),evidenced by Brd U ELISA and EDU,respectively.The results of also demonstrated that middle-concentration,high-concentration of ST increased the percentage of G2/M cells.Furhtermore,ST not only increased the expressions of cyclin D1,CDK4 and HGF protein,but also enhanced the phosphorylation level of c-met,Akt,Erk1/2(P<0.01).Conclusion: Under the conditions of this experiment,an appropriate method for the determination of phlorizin and trilobatin of ST was established in this study.ST significantly promoted the proliferation of L-02 cells,and the possible mechanism maybe due to increasing the expressions of cyclin D1,CDK4 and up-regulating HGF/c-Met signal pathway.