【摘要】 目的 研究脂联素对糖尿病患者内皮祖细胞(EPCs) 增殖、迁移能力的影响并探讨其可能机制。 方法 利用密度梯度离心法分离、培养T2DM患者外周血单个核细胞, 经FITC- UEA-I和Dil-acLDL双染色鉴定为正在分化的EPCs。将分离、培养的EPCs分为对照组、脂联素组(10ug/Ml)、PI3K抑制剂(Ly294002)干预组及ER快抑制剂(PD98059)干预组。采用MTT比色法、细胞集落形成单位计数等方法观察各组EPCs增殖能力的变化情况，应用Transwell小室法分析EPCs迁移能力的变化，同时本研究采用免疫蛋白印迹法观察脂联素处理EPCs后磷酸化Akt以及磷酸化eNOS的表达变化情况，以探讨其可能机制。 结果 与对照组相比，脂联素组EPCs的增殖能力显著提高，而在PI3K抑制剂干预组，脂联素对EPCs增殖、迁移活性的改善作用受到抑制。另外，免疫蛋白印迹法的结果显示：相比于对照组，脂联素组p-Akt、p-eNOS的表达水平明显增高，而PI3K抑制剂干预后能够有效地抑制脂联素促Akt磷酸化的效应。 结论 脂联素促进糖尿病患者EPCs增殖、迁移等功能，其主要机制可能与PI3K/Akt/eNOS的信号通路激活有关。

　　【关键词】 脂联素;内皮祖细胞;糖尿病

　　Adiponectin improving proliferation and migration of endothelial progenitor cells in vitro in T2DM patients CAO Zheng, WANG Lin, YANG Yong, et al. Department of Cardiology, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, China

　　【Abstract】Objective To study the effect and possible mechanism of adiponectin on the proliferation and migration of endothelial progenitor cells (EPCs) in vitro in T2DM patients. Methods Mononuclear cells from T2DM patients were isolated by Ficoll density gradient centrifugation and then cultured. The EPCs were identified with the FITC- UEA-I and Dil-acLDL stainings and divided into the control group, adiponectin (10ug/Ml) group, PI3K-inhibitor (Ly294002) group, and ER-inhibitor (PD98059) group. The proliferation of EPCs of all groups was observed with MTT assay, the migratory activity was calculated with colony forming units (CFU), the changes in migratory activity was analyzed with transwell assay, and the expression of phosphorylated Akt and eNOS were measured by western blotting analysis. Results Compared with the control group, the proliferative and migratory activities of EPCs were significantly enhanced after treatment in the adiponectin group, while in the PI3K-inhibitor group, the effect of adiponectin on the improvement of proliferative and migratory activities of EPCs was inhibited. The western blotting analysis showed that the expression of phosphorylated Akt and eNOS were significantly increased in the adiponectin group in comparing with the control group, but the PI3K-inhibitor could effectively inhibit the effect of adiponectin on phosphorylated Akt. Conclusion Adiponectin improves the proliferative and migratory activities of EPCs in T2DM patients by the activation of PI3K/Akt/eNOS signal pathway.

　　【Key words】 Adiponectin; Endothelial progenitor cells; Diabetes mellitus