ERK1/2和p38MAPK在介导CGRP和Ang II诱导血管平滑肌细胞Nox1表达中的作用

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (1): 13-19.

ERK1/2和p38MAPK在介导CGRP和Ang II诱导血管平滑肌细胞Nox1表达中的作用

张晓一¹，于潇华²，刘玉环²，杨丽²，王珍²，阳芳²，秦旭平²，曾泗宇³

¹长治医学院药学系药理教研室，长治 046000，山西； ²南华大学药物药理研究所血管生物学实验室，衡阳 421001，湖南； ³广东省第二人民医院药物临床试验基地，广州 510317，广东

收稿日期:2016-11-09 修回日期:2016-12-19 出版日期:2017-01-26 发布日期:2017-01-23
通讯作者: 秦旭平，男，教授，硕士生导师，研究方向：血管重构机制及药物防治。 Tel: 0734-8160781 E-mail: qinxp333@hotmail.com
作者简介:张晓一，女，本科，讲师，研究方向：心血管药理。 Tel: 0355-3151442 E-mail: zhangxiaoyi1225@aliyun.com
基金资助:
国家自然科学基金（30572192,81173060）; 广东省自然科学基金（2015A030310076）

Role of ERK1/2 and p38MAPK in the expression of Nox1 in A10 vascular smooth muscle cell induced by Ang II and CGRP

ZHANG Xiaoyi¹, YU Xiaohua², LIU Yuhuan ², YANG Li², WANG Zhen ², YANG Fang ², QIN Xuping², ZENG Siyu ³

¹Department of Pharmacy, Changzhi Medical College, Changzhi 046000, Shanxi, China; ²Laboratory of Vascular Biology, Institute of Pharmacy and Pharmacology, University of South China, Hengyang 421001, Hunan, China; ³Drug Clinical Research Center, the Second Guangdong Provincial Emergency Hospital, Guangzhou 510317, Guangdong, China

Received:2016-11-09 Revised:2016-12-19 Online:2017-01-26 Published:2017-01-23

摘要/Abstract

摘要：

目的:探讨胞外信号调节激酶1/2(ERK1/2) 和p38丝裂原激活蛋白激酶(p38MAPK)在降钙素基因相关肽（CGRP）和血管紧张素II(Ang II) 诱导血管平滑肌细胞NADPH氧化酶-1（Nox1）表达中的作用。方法: 体外培养鼠源性A10血管平滑肌细胞株（A10VSMC），用Ang II或/和CGRP作为处理因素，MTT法检测细胞增殖活力；流式细胞术检测细胞周期分布；Western blot检测ERK1/2和p38MAPK总蛋白和磷酸化蛋白以及Nox1蛋白的表达水平。结果: 与对照组和CGRP组相比，Ang II在诱导A10VSMC活力和增殖的同时，亦能增加磷酸化ERK1/2和p38MAPK信号激酶和Nox1的表达。CGRP预孵育细胞30 min 却能显著抑制Ang II诱导的细胞增殖活力、Nox1蛋白、磷酸化ERK1/2和p38MAPK的表达；二苯基碘（DPI）能抑制Ang II诱导的细胞增殖、磷酸化p38MAPK及Nox1表达，但不能抑制Ang II诱导的ERK1/2磷酸化。p38MAPK阻断剂SB203580不但能抵消Ang II诱导的Nox1表达作用，而且能协同CGRP抑制Ang II诱导的Nox1表达。而ERK1/2阻断剂PD98059对CGRP和/或Ang II诱导Nox1表达作用的影响无统计学意义。结论:Ang II诱导A10VSMC增殖与激活Nox1有关，CGRP抑制Ang II诱导的Nox1表达，可能主要通过抑制p38MAPK信号途径发挥作用，而与ERK1/2信号途径无显著关系。

关键词: 血管紧张素II, 降钙素基因相关肽, 血管平滑肌细胞, NADPH氧化酶, 丝裂原激活蛋白激酶

Abstract:

AIM: To explore the role of extracellular signal-regulated kinase1/2 (ERK1/2) and p38-Mitogen-activated protein kinase (p38MAPK) in expression of nicotinic amide adenine dinucleotide phosphate （NADPH）oxidase-1 (Nox1) in vascular smooth muscle cell (VSMC) induced by Angiotensin II (Ang II) and calcitonin gene-related peptide (CGRP). METHODS: Mouse-derived A10VSMC was cultured in vitro, the cell viability and the distribution of cell cycle was detected by MTT and flow cytometry, respectively; Western blot was used to determine the protein levels of ERK1/2, p38MAPK and Nox1. RESULTS: Compared with control or CGRP groups, the A10VSMC proliferation, protein levels of p-ERK1/2 and p-p38MAPK and Nox1 were elevated in the Ang II group, which were decreased by pretreatment of CGRP for 30 min. DPI (Nox1 inhibitor) also inhibited the A10VSMC proliferation, protein levels of p-p38MAPK and Nox1 but not p-ERK1/2 induced by Ang II. SB203580 (p38MAPK inhibitor) but not PD98059 (ERK1/2 inhibitor) enhanced the inhibitory effect CGRP on the cell proliferation and Nox1 expression induced by Ang II. CONCLUSION: The A10VSMC proliferation induced by Ang II is related to increase of Nox1 expression, and the inhibitory effect of CGRP on Nox1 expression induced by Ang II in the A10VSMC proliferation phases maybe mainly associated with p38MAPK activity but not the ERK1/2 activity.

Key words: angiotensin II, calcitonin gene related peptide, vascular smooth muscle cell, Nox, MAPK

中图分类号:

R965.2

张晓一，于潇华，刘玉环，杨丽，王珍，阳芳，秦旭平，曾泗宇. ERK1/2和p38MAPK在介导CGRP和Ang II诱导血管平滑肌细胞Nox1表达中的作用[J]. 中国临床药理学与治疗学, 2017, 22(1): 13-19.

ZHANG Xiaoyi, YU Xiaohua, LIU Yuhuan, YANG Li, WANG Zhen, YANG Fang, QIN Xuping, ZENG Siyu. Role of ERK1/2 and p38MAPK in the expression of Nox1 in A10 vascular smooth muscle cell induced by Ang II and CGRP[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(1): 13-19.

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