CGRP通过激活cAMP/PPARγ/eNOS途径改善血管内皮细胞胰岛素抵抗  

Improvement of CGRP on vascular endothelial cells insulin resistance via activation of cAMP/PPARγ/eNOS pathway

作　　者：全海燕[1,2] 刘玉环 杨丽[1] 阳芳[1] 秦旭平[1] QUAN Haiyan;LIU Yuhuan;YANG Li;YANG Fang;QIN Xuping(Laboratory of Vascular Biology,Institute of Pharmacy & Pharmacology,University of South China;Department of Pharmacology,Hunan Environmental Biology College;School of Nursing,University of South China,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学药物药理研究所血管生物学实验室 [2]湖南环境生物职业技术学院药理教研室 [3]南华大学护理学院,湖南衡阳421001

出　　处：《中国临床药理学与治疗学》2019年第6期615-622,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金项目(81173060);湖南省教育厅科学研究项目(17C1400);湖南省研究生科研创新项目(CX2017B544)

摘　　要：目的:探讨降钙素基因相关肽(CGRP)对人脐静脉内皮细胞胰岛素抵抗的保护作用及其机制。方法:用人脐静脉内皮细胞(HUVEC)建立胰岛素抵抗细胞模型(irHUVEC),分别用葡萄糖氧化酶法和蒽酮-硫酸法检测细胞葡萄糖消耗能力和糖原合成能力;RT-PCR和Westernblot分别检测目的蛋白和基因表达。结果:33.3mmol/L的高糖和5μmol/L的胰岛素处理24h可以成功诱导内皮细胞产生胰岛素抵抗。与正常HUVEC相比,irHUVEC体积变大,边界模糊,形态异常;当细胞用高糖和高胰岛素分别诱导24h和48h时,irHUVEC的葡萄糖消耗量分别减少20%和31%,并且糖原合成分别减少了55%和64%,差异都具有显著性;同时,过氧化物酶增殖体激活受体γ(PPARγ)和还原型一氧化氮合酶(eNOS)的表达均降低。CGRP可以明显增加irHUVEC葡萄糖的消耗量约20%和糖原合成增加约70%,并能增加PPARγ和eNOS蛋白及mRNA表达;SQ22536(cAMP阻断剂)能使PPARγ和eNOS的蛋白和基因表达量明显减少。结论:CGRP可以改善内皮细胞的胰岛素抵抗,其机制可能与上调环磷酸腺苷(cAMP),增加PPARγ和eNOS表达有关。AIM : To explore the protective effect and the mechanism of calcitonin gene-relatedpeptide (CGRP) on the insulin-resisted human umbilical vein endothelial cells (irHUVECs). METHODS : The irHUVECs was established using human umbilical veinendothelial cells (HUVECs), gucose oxidase (GOP-POD) method and anthrone-sulfuric acid method were employed to detect glucose consumption ability and glycogen synthesis ability, respectively. RT-PCR and Western blot were used to detect the mRNA and protein expressions of peroxisome proliferator-activated receptor gamma (PPARγ) and endothelial nitric oxide synthase (eNOS), respectively. RESULTS :The irHUVECs was successfully established when the HUVECs were treated with 33.3 mmol/L high glucose and 5 μmol/L insulin. Compared to the normal group of endothelial cells, the features of irHUVECs were a larger volume, fuzzy boundary and abnormal morphology, when cells induced were incubated with high glucose and high insulin in 24 h and 48 h, respectively, the glucose consumption was reduced by 20% and 31%, and the glycogen synthesis was reduced by 55% and 64%, meanwhile, the expressions of PPARγ and eNOS were decreased. Treatment of CGRP significantly increased about 20% glucose consumption and about 70% glycogen synthesis, and increased the expressions of PPARγ and eNOS, SQ22536 (cAMP inhibitor) can decreased the expressions of PPARγ and eNOS. CONCLUSION : CGRP could improve the endothelial cell insulin resistance, the mechanism may be related to up-regulated cAMP, and the increased expressions of PPARγ and eNOS.

关 键 词：降钙素基因相关肽 内皮细胞 胰岛素抵抗 过氧化物酶增殖体激活受体Γ 一氧化氮合酶 环磷酸腺苷 

分 类 号：R587.1[医药卫生—内分泌] R966[医药卫生—内科学]