白细胞介素-6通过AMPK/SIRT1通路下调eNOS Thr495/497

该文探讨了白细胞介素-6(interleukin-6,IL-6)对牛主动脉内皮细胞(bovine aortic endothelial cells,BAECs)的内皮型一氧化氮合成酶(endothelial nitric oxide synthase,eNOS)的影响及其可能的发生机制。在原代BAECs细胞培养基础上,选取IL-6不同浓度(10、20、40 ng/mL)、不同时间(0.5、1、2 h)处理BAECs;AMPKα1 siRNA预处理BAECs后加入IL-6共处理2 h。用Western blot方法检测BAECs细胞中eNOS、Thr497、腺苷酸活化蛋白激酶(AMPK)、p-AMPK、沉默信息调节因子2相关酶1(SIRT1)的表达情况,用一氧化氮(NO)检测试剂盒检测BAECs细胞中NO含量。结果显示,与正常对照组相比,IL-6显著降低Thr497的表达(P<0.05),同时伴有p-AMPK、SIRT1上调(P<0.05),NO含量升高。而IL-6对Thr497的下调作用被AMPKα1 siRNA和EX-527阻断(P<0.05)。总之,IL-6可下调BAECs中Thr497表达进而影响eNOS活性,其作用机制可能涉及到AMPK/SIRT1通路的参与。

Interleukin-6 Down-Regulates eNOS Thr495/497 through AMPK/SIRT1 Pathway

This is a study on the effects of IL-6(interleukin-6)on eNOS(endothelial nitric oxide synthase)of BAECs(bovine aortic endothelial cells)and its mechanism.BAECs were treated with IL-6 in different concentrations(10,20,40 ng/mL)and different times(0.5,1,2 h).Beside,BAECs were pre-treated with AMPKα1 siRNA and then were treated with IL-6 for 2 h(kept with AMPKα1 siRNA).The expression of eNOS,Thr497,AMPK(adenylate-activated protein kinase),p-AMPK,and SIRT1(silence information regulator 2 related enzyme 1)in BAECs were detected by Western blot.The NO(nitric oxide)content in BAECs was measured by NO detection kit.The results showed that IL-6 significantly reduced the expression of Thr497(P<0.05),accompanied by up-regulation of p-AMPK,SIRT1(P<0.05),and increased NO content compared with the normal control group.The down-regulation of Thr497 by IL-6 was blocked by AMPKα1 siRNA and EX-527(P<0.05).In short,IL-6 could down-regulate the expression of Thr497 in BAECs and then affect eNOS activity.Its mechanism may involve the participation of AMPK/SIRT1 pathway.