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MCPIP1介导MCP-1诱导的血管平滑肌细胞增殖

杨一峻, 吴柳松, 束波, 钱民章^*

贵州省遵义医学院生物化学教研室,遵义 563003

摘要

探讨MCPIP1 是否介导 MCP-1 诱导的血管平滑肌细胞（vascular smooth muscle cell， VSMC）增殖。采用不同剂量的MCP-1（1、10、100 ng/mL）分别孵育VSMC 24 h； 100 ng/mL MCP-1 分别孵育VSMC 1、4、8、12、24 h，用Real-time PCR 及 Western blot 检测MCPIP1 表达。应用MCPIP1 siRNA 沉默hVSMC 中MCPIP1 基因，100 ng/mL MCP-1 孵育VSMC，细胞计数板记录细胞数量变化；CCK-8 试剂盒检测细胞活力的变化；EdU 试剂盒检测VSMC DNA合成率；流式细胞术检测S期细胞数量；用100 ng/mL MCP-1 或20ng/mL血小板衍生生长因子（platelet-derived growth factor, PDGF）孵育VSMC 后，Real-time PCR 检测c-fos 基因mRNA 的转录。结果显示：1、10、100 ng/mL MCP-1均能诱导VSMC 中MCPIP1 mRNA 和蛋白的表达（P<0.01），并存在剂量依赖效应。100 ng/mL MCP-1孵育VSMC 4 h后， MCPIP1表达与对照组相比有明显增高（P<0.01），并存在时间依赖效应。用100 ng/mL MCP-1 孵育VSMC 后，MCPIP1 基因沉默组与对照组比较，细胞数量、细胞活力及细胞DNA合成率均明显降低（P<0.01），S 期细胞数量下降（P<0.05）。沉默MCPIP1基因后， MCP-1、PDGF诱导VSMC 中c-fos 基因表达下调，与对照组比较差异均有统计学意义 （P<0.01）。结果提示：MCPIP1 介导了MCP-1 诱导的VSMC 增殖，并可能与诱导 c-fos 表达有关。

关键词： 单核细胞趋化蛋白-1; 单核细胞趋化蛋白-1诱导蛋白1; 平滑肌细胞增殖; c-fos基因

[MCPIP1 mediates MCP-1-induced vascular smooth muscle cell proliferation.] [Article in Chinese]

YANG Yi-Jun, WU Liu-Song, SHU Bo, QIAN Min-Zhang^*

Department of Biochemistry, Zunyi Medical College, Zunyi 563003, China

Abstract

The aim of the present study is to investigate whether monocyte chemotactic protein-1 (MCP-1)-induced vascular smooth muscle cell (VSMC) proliferation is mediated via monocyte chemotactic protein-1 induced protein-1 (MCPIP1). MCPIP1 expressions in cultured VSMC were detected by real-time PCR and Western blot following MCP-1 incubation. After MCPIP1 was silenced by siRNA, cell number was counted by hemocytometer, VSMC activity was analyzed by CCK-8 kit, percentage of DNA synthesis was detected by EdU kit, percentage of S phase cell numbers were measured by flow cytometry, and c-fos mRNA expression induced by MCP-1 or platelet-derived growth factor (PDGF) was detected by real-time PCR. The results showed MCP-1 increased MCPIP1 mRNA and up-regulated MCPIP1 protein expression in dose- and time-dependent manners. Cell counts, cellular activity, the percentage of DNA synthesis, and the percentage of S phase cell numbers were remarkably decreased in MCPIP1 siRNA group, compared with those in MCP-1 group. The enhancing effect of MCP-1 or PDGF on c-fos mRNA expression was inhibited by MCPIP1 siRNA. These results suggest that MCP-1-induced VSMC proliferation is mediated via MCPIP1, and the underlying mechanism may involve c-fos expression up-regulation.

Key words: monocyte chemotactic protein-1; monocyte chemotactic protein-1 induced protein-1; vascular smooth muscle cell proliferation; c-fos

收稿日期：　　录用日期：

通讯作者：钱民章　　E-mail: qian_mzh@hotmail.com

引用本文：

杨一峻, 吴柳松, 束波, 钱民章. MCPIP1介导MCP-1诱导的血管平滑肌细胞增殖[J]. 生理学报 2013; 65 (6): 616-622.

YANG Yi-Jun, WU Liu-Song, SHU Bo, QIAN Min-Zhang. [MCPIP1 mediates MCP-1-induced vascular smooth muscle cell proliferation.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (6): 616-622 (in Chinese with English abstract).