腺苷酸蛋白激酶活化在红景天苷抑制同型半胱氨酸诱导的人脐静脉内皮细胞内质网应激中的作用  

Salidroside Inhibits Homocysteine-Induced Endoplasmic Reticulum Stress through Activation of AMP-Activated Protein Kinase

作　　者：俞天虹[1] 朱琳[1] 赵蓉[1] 王彦军[1] 孙建辉[1] 贾方[1] 

机构地区：[1]苏州大学附属第三医院心内科,江苏常州213003

出　　处：《中华中医药学刊》2017年第10期2558-2562,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81300220);江苏省自然科学基金项目(BK20140251)

摘　　要：目的:探讨腺苷酸蛋白激酶(AMPK)活化在红景天苷(Sal)抑制同型半胱氨酸(Hcy)诱导的脐静脉内皮细胞(HUVECs)内质网应激中的作用。方法:将AMPK基因的siRNA载体(AMPK-si-RNA)转染HUVECs后,用红景天苷或AMPK的选择性激动剂5-氨-4-甲酰胺咪唑核糖核苷酸(AICAR)预孵育2 h,再加入Hcy处理24 h,实时荧光定量PCR(Realtime-PCR)检测免疫球蛋白结合蛋白(Bi P)、C/EBP同源蛋白(CHOP)的mRNA表达水平,Western Blot检测Bi P、CHOP、AMPK的蛋白表达变化,双链RNA依赖的PKR内质网激酶(PERK)、翻译起始因子(e IF2α)和AMPK的磷酸化水平。结果:与空白对照组相比,单用Hcy处理组的Bi P和CHOP基因表达和蛋白表达都显著升高(P<0.01),PERK和e IF2α的磷酸化水平升高(P<0.01),与单用Hcy处理组相比,红景天苷和AICAR预处理组均能抑制Hcy导致的Bi P和CHOP的基因和蛋白表达(P<0.05),降低PERK和e IF2α的磷酸化水平(P<0.05),而AMPK-si-RNA转染细胞后,红景天苷抑制Bi P和CHOP基因和蛋白表达的能力下降,抑制PERK和e IF2α的磷酸化水平的能力也相应降低。结论:红景天苷抑制Hcy诱导的内质网应激是由AMPK活化介导的,从而发挥内皮保护的效应。Objective： To investigate the effect of Salidroside （Sal） on homocysteine （Hcy） induced endoplasmic reticulum stress in human umbilical vein endothelial cells （HUVECs） and the potential mechanisms regarding AMP - activated protein kinase （AMPK）. Methods ： HUVECs were transfected with AMPK - si - RNA, pretreated with Salidroside or AICAR for 2 hours, and then incubated with Hcy for 24 hours. The mRNA levels of BiP and CHOP were detected by realtime PCR. The protein levels of BiP, CHOP, AMPK, p - AMPK, p - PERK and p - eIF2α were examined by Western blot. Result：Compared with the normal group, the BiP and CHOP mRNA expression and protein expression were significantly increased in the Hcy treated group （P 〈 0.01 ） and the phosphorylation levels of PERK and eIF2 were elevated （P 〈 0. 01 ）. Compared with the Hcy treatment group, the mRNA and protein expressions of BiP and CHOP were inhibited in Salidroside and AICAR treatment group （ P 〈 0. 05 ）, which decreased the phosphorylation levels of PERK and eIF2 （ P 〈 0. 05）. After transfected HUVECs by AMPK - si - RNA, the ability of Salidroside to inhibit the mRNA and protein expression of BiP and CHOP were decreased and the capability to inhibit the phosphorylation level of PERK and eIF2α was also faded accordingly. Conclusion：The inhibitory effect of Hcy on the endoplasmic reticulum stress induced by Salidroside is mediated by the activation of AMPK, which exerts protective effect on the endothelium.

关 键 词：腺苷酸蛋白激酶 红景天苷 同型半胱氨酸 内质网应激 

分 类 号：R285.5[医药卫生—中药学]