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本文旨在探讨单核细胞趋化蛋白-1诱导蛋白1(monocyte chemotactic protein-1 induced protein-1,MCPIP1)是否介导单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)诱导的血管平滑肌细胞(vascular smooth muscle cell,VSMC)增殖。用Real-time PCR及Western blot检测体外培养的VSMC中MCP-1诱导的MCPIP1表达;用siRNA沉默VSMC中MCPIP1基因后,用细胞计数板记录细胞数量,用CCK-8试剂盒检测细胞活力,用EdU试剂盒检测DNA合成率,用流式细胞术检测S期细胞数量,用Real-time PCR检测MCP-1或血小板衍生生长因子(platelet-derived growth factor,PDGF)诱导的c-fos mRNA水平变化。结果显示,MCP-1提高VSMC中MCPIP1 mRNA水平,并呈剂量和时间依赖性地上调MCPIP1蛋白表达。和MCP-1处理的VSMC相比,MCPIP1基因沉默后VSMC的细胞数量、细胞活力及细胞DNA合成率均明显降低,S期细胞数量下降;MCPIP1基因沉默可抑制MCP-1或PDGF对c-fos mRNA的上调作用。以上结果提示,MCPIP1介导了MCP-1诱导的VSMC增殖,其机制涉及对c-fos表达的上调。
This study aimed to investigate whether monocyte chemotactic protein-1 induced protein-1 (MCPIP1) mediates monocyte chemotactic protein-1 (MCP-1) Induced proliferation of vascular smooth muscle cells (VSMCs). MCP-1-induced MCPIP1 expression in VSMCs cultured in vitro was detected by Real-time PCR and Western blot. After silencing the MCPIP1 gene in VSMCs with siRNA, the cell numbers were recorded with a cell counting plate. Cell viability was measured by CCK-8 kit. EdU kit was used to detect the DNA synthesis rate. The number of S phase cells was detected by flow cytometry. The changes of c-fos mRNA level induced by MCP-1 or platelet-derived growth factor (PDGF) were detected by Real-time PCR . The results showed that MCP-1 increased the level of MCPIP1 mRNA in VSMCs and up-regulated the expression of MCPIP1 protein in a dose-and time-dependent manner. Compared with MCP-1 -treated VSMC, the number of cells, cell viability and DNA synthesis in VSMCs were significantly decreased and the number of cells in S phase decreased after MCPIP1 gene silencing. MCPIP1 gene silencing could inhibit the expression of c-fos Upregulation of mRNA. These results suggest that MCPIP1 mediates the proliferation of VSMCs induced by MCP-1, and its mechanism involves the up-regulation of c-fos expression.