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目的探讨促生长激素释放肽(Ghrelin)促进人脐静脉内皮细胞(HUVEC)增殖的分子机制。方法将HUVEC分为正常对照(Con)组、Ghr组、D-Lys-3-GHRP组、Ghr+D-Lys-3-GHRP组、雷帕霉素(Rapa)组及Ghr+Rapa组,采用MTT法检测细胞增殖能力,Western blot检测HUVEC中Ghrelin受体(GHSR1a)的表达及哺乳类雷帕霉素靶蛋白(mTOR)、p70S6K和S6的磷酸化水平。结果与Con组相比,MTT检测显示Ghr组细胞增殖增强,且高于Ghr+D-Lys-3-GHRP组和Ghr+Rapa组;Western blot检测显示,与Con组比较,mTOR、p70S6K和S6在Ghr组快速磷酸化,Ghr+D-Lys-3-GHRP组mTOR、P70S6K和S6磷酸化水平低于Ghr组。结论 Ghrelin与其受体GHSR1a结合后,活化mTOR/p70S6K信号通路,促进HUVEC增殖。
Objective To investigate the molecular mechanism of Ghrelin promoting the proliferation of human umbilical vein endothelial cells (HUVECs). Methods HUVECs were divided into normal control group, Ghr group, D-Lys-3-GHRP group, Ghr + D-Lys-3-GHRP group, Rapa group and Ghr + Rapa group The cell proliferation was detected by MTT assay. The expression of Ghrelin receptor (GHSR1a) and the phosphorylation of mammalian target proteins of rapamycin (mTOR), p70S6K and S6 were detected by Western blot. Results Compared with Con group, MTT assay showed that the proliferation of Ghr group was higher than that of Ghr + D-Lys-3-GHRP group and Ghr + Rapa group. Western blot showed that compared with Con group, mTOR, p70S6K and S6 In the Ghr group, the phosphorylation of mTOR, P70S6K and S6 in Ghr + D-Lys-3-GHRP group was lower than that in Ghr group. Conclusion After Ghrelin binding with its receptor GHSR1a, it activates the mTOR / p70S6K signaling pathway and promotes the proliferation of HUVECs.