动物造模 z-bio 2021-07-13 277

　　Objective: To target rapamycin to inhibit mTOR and study its effect on the proliferation of rat heart valve interstitial cells.

　　Method: In vitro isolation, culture and identification of rat valve stromal cells. The cells were randomly divided into two groups: rapamycin group and control group. The MTT method was used to detect and draw the growth curve of rat valve interstitial cells. Post-processing; flow cytometry to detect the effect of rapamycin on cell cycle; RT-PCR to detect the expression level of S6 and P70 S6K mRNA in cells; to detect the expression level of S6 in S6, P70S6K and P cells Western blotting and P-P70S6K protein .

　　Result: We successfully isolated and cultured rat valve stromal cells isolated in vitro. The cell activity of the control group was significantly higher than that of the drug-added group (Pu003c0.05). The cell activity of the drug flow cytometer showed that the proportion of cells in the drug-added group had no significant difference. In each cycle of the control group, it showed that the drug-added group S6 and P70 The phosphorylation level of S6K protein is reduced (P). u003c0.05).

　　Conclusion: Rapamycin can inhibit the proliferation of valve interstitial cells. The reason may be that the phosphorylation levels of target proteins S6 and P70S6K are down-regulated after mTOR inhibition, rather than cell cycle changes.