The Effects Of Tetramethylpyrazine In Combination With Rhein Oninjuried Hepatocytes Induced By TGF-β₁ And TG

Objective To investigate the effects of Tetramethylpyrazine(TMP) in combination with Rhein(RH) on injuried human liver cell line L-02 induced by TGF-β1 and TG. Methods Tetramethylpyrazine and Rhein was added to the L-02 cells cultured medium with different concentration to decide the suitable concentrations of the drugs. To create the injuried hepatocyte model the combination of the TGF-β1 and TG was used. Then TMP in combination with RH was added to the injured hepatocyte model.To investigate the effects the drugs ,the MTT method was used to assay the cellproliferation, the flow cytometic analysis was used to assay the cell apoptosis, the morphological changes of the apoptotic L-02 cells stained with AO-EB was observed by the fluorescence microscope and the expressin of the PCNA,Bcl-2,Bax and Caspase3 were detected with immunocytochemical method. Result Tetramethylpyrazine (10 μg/ml,50μg/ml) and Rhein(1μg/ml , 5 μ g/ml)have the interruption effect on the inhibition of cellproliferation induced by the TGF-β1 and TG. Tetramethylpyrazine(10μ g/ml,50 μg/ml) and Rhein(5μg/ml) have the transparent inhibition on the L-02 cells apoptosis induced by the TGF-β1 and TG. TMP in combination with RH(TMP 10μg/ml and RH 5μg/ml,1:1) could block the inhibition of the cellproliferation and inhibit the cell apoptosis.The drugs could up-regulate the expression of PCNA and Bcl-2, down-regulate the expression of Bax and Caspase3. The combination of the drugs had more obvious protective effect than the mono-drugs. Conclusion Tetramethylpyrazine and Rhein have the protective effect on the injuried hepatocyte induced by the TGF-β1 and TG. And the effect of TMP in combination with RH was more significant than TMP and RH. Blocking the inhibition of the cellproliferation, up-regulating the expression of Bcl-2 and down-regulating the expression of Bax and Caspase3 would be the mechanism of the protective effect.