动物实验,雷帕霉素 z-bo 2020-08-22 462

　　Objective: To inhibit mTOR with rapamycin as a target, and study its effect on the proliferation of rat heart valve stromal cells.

　　Method: Isolate, culture and identify rat valve stromal cells in vitro; divide the cells into two groups randomly: rapamycin group and control group; before and after treatment with MTT method. We detected and plotted the growth curve of rat valve stromal cells. Flow cytometry was used to detect the effect of rapamycin on the cell cycle; T-PCR was used to detect the expression levels of S6 and P70S6K mRNA in the cells. Western blotting is used to detect the expression levels of S6, P70S6K, P-S6 and P-P70S6K proteins in cells.

　　Result: We successfully isolated and cultured rat valve interstitial cells isolated in vitro; the cell activity of the control group was significantly higher than that of the drug-added group (P\u003c0.05), and the proliferation was slow. The results of flow cytometry showed that there was no significant difference in the percentage of cells per cycle between the drug-added group and the control group; the phosphorylation levels of S6 and P70S6K proteins in the drug-added group decreased (P\u003c0). .05).

　　Conclusion: Rapamycin may inhibit the proliferation of valve stromal cells. The reason may be due to the suppression of mTOR and the down-regulation of phosphorylation levels of its target proteins S6 and P70S6K rather than cell cycle changes.