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目的探讨腺苷酸活化蛋白激酶(AMPK)对同型半胱氨酸(Hcy)诱导的人脐静脉内皮细胞(HUVECs)内质网应激信号通路的影响。方法应用AMPK的选择性激活剂5-氨基-4-甲酰胺咪唑核糖核苷酸(AICAR)或AMPK显性失活突变体(AMPK-DN)预处理HUVECs,RT-PCR和Western blot分别检测Hcy孵育HUVECs后,双链RNA依赖的蛋白激酶样内质网激酶(PERK)、翻译起始因子2α(eIF2α)的磷酸化活化,分子信使葡萄糖调节蛋白78(GRP78)以及凋亡信号CCAAT/增强子结合蛋白同源蛋白(CHOP)、半胱天冬氨酸蛋白酶12(Caspase-12)的mRNA和蛋白表达变化。结果 AICAR可抑制Hcy诱导的HUVECs内质网应激信号分子的表达;AMPK-DN拮抗了AICAR的抑制效应。结论 AMPK可通过调节内质网应激相关信号通路抑制Hcy诱导的内皮损伤,发挥内皮保护效应。
Objective To investigate the effect of AMPK on homocysteine (Hcy) -induced ER stress signaling pathway in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were pretreated with AICAR or AMPK-DN, a selective activator of AMPK. The expression of Hcy was detected by RT-PCR and Western blot, respectively Double-stranded RNA-dependent protein kinase-like endoplasmic reticulum kinase (PERK), phosphorylation of translation initiation factor 2α (eIF2α), molecular messenger glucose regulatory protein 78 (GRP78) and the apoptotic signal CCAAT / enhancer (CHOP) and Caspase-12 mRNA and protein expression in the mice. Results AICAR inhibited the expression of Hcy-induced endoplasmic reticulum stress signaling molecules in HUVECs. AMPK-DN antagonized the inhibitory effect of AICAR. Conclusion AMPK can inhibit endothelium injury induced by Hcy by regulating endoplasmic reticulum stress-related signaling pathway and exert endothelial protective effect.