磷酸腺苷激活的蛋白激酶对抵抗素诱导内皮细胞凋亡的保护作用研究

Protective effects of AMPK on resistin induced human umbilical vein endothelial cell apoptosis

目的研究抵抗素对内皮细胞功能的影响和磷酸腺苷激活的蛋白激酶(AMPK)在抵抗素影响内皮功能中的作用。方法原代培养人脐静脉内皮细胞(HUVEC),不同浓度人抵抗素(0、50和100ng/mL)培养24h。TUNEL和Annexin V-FITC分别检测其对内皮细胞凋亡的影响,H2DCFDA染色检测细胞内活性氧簇(ROS)生成,Western-blot检测内皮细胞AMPK-α(Thr172)磷酸化水平。抵抗素干预后,采用AMPK激动剂AICAR干预观察AMPK在抵抗素影响内皮细胞中的作用。结果抵抗素干预内皮细胞并不能增加其凋亡和ROS生成,但可抑制内皮细胞AMPK的磷酸化激活,且AMPK的激活能够抑制抵抗素作用的内皮细胞凋亡和ROS生成。结论抵抗素对内皮细胞的作用机制中AMPK可能占重要地位,抵抗素可抑制内皮AMPK磷酸化,而AICAR激活AMPK后对内皮细胞损伤起保护作用。

[Objective] To investigate the effects of resistin on endothelial cell, and to explore the role of AMPK in the endothelial cell dysfunction affected by resistin. [Methods] Human umbilical vein endothelial cells were incubated with recombinant human resistin （0, 50 and 100 ng/mL） for 24 hours. Apoptosis and ROS production and AMPK phosphorylation of endothelial cell were assessed. For the observation of the role of AMPK, AICAR was used for the activation of AMPK in resitin treated cells and apoptosis, ROS production and AMPK phosphorylation were assessed too. [Results] Incubation of HUVEC with resistin did not result in the alter of cell ROS production and apoptosis, but activation of AMPK in endothelial cell treated by resistin induced a decreasing of ROS production and cell apoptosis. [Conclusions] AMPK pathway play an important role in resistin treated endothelial cell, the activation of AMPK may have the protective effects of endothelial cell dysfunction.