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

doi:10.12092/j.issn.1009-2501.2019.06.003

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (6): 615-622.doi: 10.12092/j.issn.1009-2501.2019.06.003

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

全海燕^1,2，刘玉环³，杨丽¹，阳芳¹，秦旭平¹

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，衡阳 421001，湖南

收稿日期:2018-11-02 修回日期:2019-04-16 出版日期:2019-06-26 发布日期:2019-06-25
通讯作者: 秦旭平，男，博士，硕士研究生导师，研究方向：心血管药理学。 Tel：0734-8160781 E-mail:522754837@qq.com
作者简介:全海燕，女，硕士，研究方向：心血管药理学。 E-mail:441348824@qq.com 刘玉环，女，实验师，本文同等第一作者。 E-mail:499035589@qq.com
基金资助:
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)

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

QUAN Haiyan^1,2, LIU Yuhuan³, YANG Li¹, YANG Fang ¹, QIN Xuping ¹

¹ Laboratory of Vascular Biology, Institute of Pharmacy & Pharmacology, University of South China, ²Department of Pharmacology, Hunan Environmental Biology College, 3School of Nursing, University of South China, Hengyang 421001, Hunan,China

Received:2018-11-02 Revised:2019-04-16 Online:2019-06-26 Published:2019-06-25

摘要/Abstract

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Western blot分别检测目的蛋白和基因表达。结果:33.3 mmol/L的高糖和5 μmol/L 的胰岛素处理24 h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24 h和48 h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。

Abstract:

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.

Key words: calcitonin gene related peptide, human umbilical vein endothelial cells, insulin resistance, PPARγ, eNOS, cAMP

中图分类号:

R587.1
R966

全海燕，刘玉环，杨丽，阳芳，秦旭平. CGRP通过激活cAMP/PPARγ/eNOS途径改善血管内皮细胞胰岛素抵抗[J]. 中国临床药理学与治疗学, 2019, 24(6): 615-622.

QUAN Haiyan, LIU Yuhuan, YANG Li, YANG Fang, QIN Xuping. Improvement of CGRP on vascular endothelial cells insulin resistance via activation of cAMP/PPARγ/eNOS pathway[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(6): 615-622.

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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

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