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受体成分蛋白在降钙素基因相关肽和血管紧张素II对血管平滑肌细胞血管过氧化物酶1表达调控中的作用

刘彦梅, 彭虹艳, 郭锋, 全海燕, 骆镜妃, 秦旭平^*

南华大学药物药理研究所，湖南省分子靶标新药研究协同创新中心，衡阳 421001

摘要

血管紧张素II (angiotensin II, Ang II)和降钙素基因相关肽(calcitonin gene-related peptide, CGRP)在血管的损伤和保护中起重要作用。为了探讨CGRP受体成分蛋白(receptor component protein, RCP)在CGRP和Ang II对血管平滑肌细胞(vascular smooth muscle cell, VSMC)血管过氧化物酶1 (vascular peroxidase-1, VPO1)表达调控中的作用及机制，本研究采用体外培养鼠源性A10血管平滑肌细胞株(A10VSMC)，给予CGRP或/和Ang II刺激，并用小干扰RNA (siRNA)干扰细胞RCP基因的表达，Western Blot检测RCP以及VPO1蛋白表达水平；RT-PCR检测RCP及VPO1 mRNA的表达水平。结果显示，在静止期野生型A10VSMC，CGRP和Ang II能分别上调RCP和VPO1蛋白和mRNA表达(均P < 0.05)，但CGRP预孵育细胞后，Ang II诱导的RCP和VPO1蛋白表达降低(均P < 0.05)；与野生型组比较，VPO1在所有RCP基因干扰组的表达均显著降低(均P < 0.01)。同时，在RCP基因干扰条件下，和对照组相比，CGRP处理组VPO1蛋白的表达显著增加，而Ang II组没有明显变化；和Ang II组相比，CGRP与Ang II联合作用显著增加VPO1蛋白表达，但这种作用能被抗氧化酶Catalase所抑制(P < 0.05)。以上结果提示，RCP可能参与CGRP或Ang II诱导的VPO1蛋白表达；RCP可能在介导CGRP和Ang II受体共同调控VPO1表达的信号转导整合中起一定作用。

关键词： 降钙素基因相关肽; 受体成分蛋白; 血管紧张素II; 血管过氧化物酶1; 血管平滑肌细胞

分类号：R361.3

[Involvement of the receptor component protein in the regulation of vascular peroxidase-1 expression induced by calcitonin gene-related peptide and angiotensin II in vascular smooth muscle cell.] [Article in Chinese]

LIU Yan-Mei, PENG Hong-Yan, GUO Feng, QUAN Hai-Yan, LUO Jing-Fei, QIN Xu-Ping^*

Institute of Pharmacy and Pharmacology, Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study, University of South China, Hengyang 421001, China

Abstract

Angiotensin II (Ang II) and calcitonin gene-related peptide (CGRP) play important roles in vascular injury and protection. In order to determine the role of CGRP receptor component protein (RCP) in signal transduction whereby CGRP and Ang II mediate the expression of vascular peroxidase-1 (VPO1) in vascular smooth muscle cell (VSMC), mouse derived A10 vascular smooth muscle cell line (A10VSMC) was cultured with CGRP or/and Ang II in vitro. RCP-specific small interference RNA (siRNA-RCP) was used to silence oligonucleotide sequence. Western blot and RT-PCR were used to determine the protein and mRNA expressions of RCP and VPO1, respectively. The results showed that the expressions of RCP and VPO1 were increased in the presence of CGRP or Ang II in the quiescent A10VSMC. But the protein expressions of RCP and VPO1 induced by Ang II were decreased by pretreatment of CGRP (P < 0.05). The expressions of VPO1 were decreased in all the groups treated with siRNA-RCP, compared with those of wide-type counterparts. Meanwhile, the expression of VPO1 was significantly induced by CGRP but not Ang II in the siRNA-RCP treated A10VSMCs. Ang II in combination with CGRP increased the protein expression of VPO1 in the siRNA-RCP-transfected cells, compared with Ang II alone, and this effect could be abolished by catalase. The results suggest that RCP may play an important role in the integration of signal transduction whereby CGRP and Ang II receptors jointly regulate VPO1 expression in VSMC.

Key words: Calcitonin gene-related peptide; calcitonin gene-related peptide receptor component protein; angiotensin II; vascular peroxidase-1; Vascular smooth muscle cell

收稿日期：2014-10-23　　录用日期：2014-12-31

通讯作者：秦旭平　　E-mail: qinxuping@sohu.com

引用本文：

刘彦梅, 彭虹艳, 郭锋, 全海燕, 骆镜妃, 秦旭平. 受体成分蛋白在降钙素基因相关肽和血管紧张素II对血管平滑肌细胞血管过氧化物酶1表达调控中的作用[J]. 生理学报 2015; 67 (2): 193-200.

LIU Yan-Mei, PENG Hong-Yan, GUO Feng, QUAN Hai-Yan, LUO Jing-Fei, QIN Xu-Ping. [Involvement of the receptor component protein in the regulation of vascular peroxidase-1 expression induced by calcitonin gene-related peptide and angiotensin II in vascular smooth muscle cell.] [Article in Chinese]. Acta Physiol Sin 2015; 67 (2): 193-200 (in Chinese with English abstract).