动物造模 z-bio 2023-02-02 372

　　OBJECTIVE: To use rapamycin to target mTOR and investigate its effect on the proliferation of rat heart valve interstitial cells.

　　Methods: Rat valve interstitial cells were isolated in vitro, cultured and identified; cells were randomly divided into two groups: rapamycin group and control group; MTT method was used to detect and draw the growth curve of rat valve interstitial cells before and after drug administration; flow cytometry The effect of rapamycin on cell cycle was detected by cytometer; the mRNA expression levels of S6 and P70S6K in cells were detected by RT-PCR; the protein expression levels of S6, P70S6K, P-S6 and P-P70S6K in cells were detected by Western blotting.

　　RESULTS: The rat valve interstitial cells isolated in vitro were successfully isolated and cultured; the cell activity in the control group was significantly higher than that in the drug-added group (P<0.05), and the growth of the cells in the drug-added group was slowed down. Flow cytometry showed that there was no significant difference in the proportion of cells in each cycle between the drug-added group and the control group; the phosphorylation levels of S6 protein and P70S6K protein in the drug-added group were decreased (P<0.05).

　　Conclusion: The reason why rapamycin can inhibit the proliferation of valvular interstitial cells may not be caused by changing the cell cycle but by down-regulating the phosphorylation levels of its target proteins S6 and P70S6K after inhibiting mTOR.