肢体缺血再灌注损伤 z-bio 2021-07-13 632

　　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, P70 S6K and P cells Western blotting and P- The P70S6K protein is for.

　　Result: Rat valve stromal cells isolated in vitro were successfully isolated and cultured. The cell activity of the control group was significantly higher than that of the drug-added group (Pu003c0.05). The cell activity of drug flow cytometry showed that there was no significant difference in the proportion of cells in the drug-added group. In each cycle of the control group, the drug-added groups S6 and P70 showed a decrease in S6K protein phosphorylation (P). u003c0.05). \nConclusion: 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. \n Purpose: To investigate the changes of platelet mitochondria after the tourniquet ischemia-reperfusion injury, and to provide guidance for the clinical application of the tourniquet.

　　Method: 30 SD rats were randomly divided into a control group (6 rats) and an ischemia-reperfusion group (24 rats, then randomly divided into 4 groups, and blood was collected at 2, 6, 12, and 24 hours later.). Reperfusion. Detection). The root of the thigh of the white rabbit (pressure 280 mmHg) was wrapped with a self-made tourniquet to simulate ischemia of the tourniquet, and the tourniquet was released 4 hours later. At the corresponding time point, blood is drawn to separate platelets. Use automatic platelet counter to measure platelet count; use luciferin-luciferase kit to measure platelet ATP content; use JC-1 mitochondrial membrane potential kit (△ ym) to detect changes in platelets; use cytochrome C apoptosis kit to detect platelets The content of cytochrome C; lipid peroxide kit for detecting lipid peroxide in platelets.

　　Result: Compared with the control group, the platelet ATP content of the ischemia-reperfusion injury group significantly decreased at 2 hours and 6 hours (P\u003c0.05). There was no statistically significant difference in platelet count between the two groups (P\u003c0.05).

　　Conclusion: Tourniquet ischemia-reperfusion injury can lead to impaired platelet mitochondrial function, which mainly occurs in the early stage of ischemia-reperfusion (6 hours) and gradually recovers in the later stage.