富血小板血浆促进人子宫内膜间充质干细胞(EnMSCs)增殖的机制

Platelet-rich Plasma Promotes the Proliferation of Human Endometrial Mesenchymal Stem Cells(EnMSCs)through the PI3K/AKT/mTOR Signaling Pathway

目的探讨PRP促进EnMSCs增殖的机制,为EnMSCs的扩增及临床应用提供理论基础。方法将EnMSCs随机分为对照组、2%PRP组、2%PRP+LY294002组,CCK-8法检测细胞增殖情况;流式细胞仪监测细胞周期及Western Blot和ELISA检测细胞中p-PI3K、AKT、p-AKT、mTOR及p-mTOR的蛋白表达。结果(1)CCK-8结果显示:与对照组相比较,2%PRP组EnMSCs的增殖水平显著较高(P<0.05);与2%PRP组相比,2%PRP+LY294002组EnMSCs的增殖水平显著较低(P<0.05);(2)流式细胞仪分析细胞周期结果显示:2%PRP组G2/M期细胞比例显著高于对照组和2%PRP+LY294002组,G0/G1细胞比例明显低于对照组和2%PRP+LY294002组,差异均有统计学意义(P<0.05);(3)2%PRP组EnMSCs中p-PI3K、AKT、p-AKT、mTOR、p-mTOR的蛋白表达明显高于对照组及2%PRP+LY294002组,差异均有统计学意义(P<0.05)。结论EnMSCs的增殖由PI3K/AKT/mTOR信号通路调控,PRP通过促进AKT、mTOR的蛋白表达及其磷酸化以激活该通路,促进EnMSCs由G1期向G2期转变,从而促进EnMSCs的增殖。

Objective To explore the mechanism of PRP promoting EnMSCs proliferation,and to provide a theoretical basis for the expansion and clinical application of EnMSCs.Methods EnMSCs were randomly divided into the control group,2%PRP group and 2%PRP+LY294002 group.Cell proliferation was detected by CCK-8 method.Cell cycle was monitored by flow cytometry and protein expressions of p-PI3 K,AKT,p-AKT,mTOR and p-mTOR were detected by Western Blot.Results(1)CCK-8 results:Compared with the control group,the proliferation level of EnMSCs in 2%PRP group was significantly higher(P<0.05).Compared with the 2%PRP group,the proliferation level of EnMSCs in the 2%PRP+LY294002 group was significantly lower(P<0.05).(2)The results of cell cycle analysis by flow cytometry showed that the proportion of cells in G2/M phase in the 2%PRP group was significantly higher than that in the control group and 2%PRP+LY294002 group,and the proportion of cells in G0/G1 phase was significantly lower than that in the control group and 2%PRP+LY294002 group.All of these had statistically significant differences(P<0.05).(3)The protein expressions of p-PI3 K,AKT,p-AKT,m TOR and p-m TOR in En MSCs in the 2%PRP group were significantly higher than those in the control group and the 2%PRP+LY294002 group,with statistical significance(P<0.05).Conclusion The proliferation of En MSCs is regulated by the PI3 K/Akt/m TOR signaling pathway.PRP activates this pathway by promoting the protein expression and phosphorylation of AKT and mTOR,thereby promoting the proliferation of EnMSCs.