细胞外基质金属蛋白酶诱导子突变的血管平滑肌细胞的构建及其功能研究
Construction and functional study of vascular smooth muscle cells with extracellular matrix metalloproteinase inducer mutation
张云燕 1林威钢 2徐程 1张芸 1方崇峰1
作者信息
- 1. 317000 临海,温州医科大学附属台州医院心血管内科
- 2. 317000 临海,温州医科大学附属台州医院整形科
- 折叠
摘要
目的 探讨细胞外基质金属蛋白酶诱导子(EMMPRIN)c.889C>A点突变对血管平滑肌细胞增殖、迁移及炎症反应等功能的影响.方法 利用CRISPR/Cas9 技术构建EMMPRIN c.889C>A点突变血管平滑肌细胞系;Real-time qPCR和Western blot实验检测EMMPRIN的表达水平;CCK-8 实验检测细胞增殖活性;Transwell实验检测细胞迁移能力;酶联免疫吸附法检测白细胞介素6(IL-6)、巨噬细胞迁移因子1(MCP-1)及白细胞介素1β(IL-1β)的分泌.结果 与野生型细胞相比,c.889C>A突变细胞中EMMPRIN的表达水平无统计学意义(t=0.732,P>0.05),细胞增殖活性差异无统计学意义(P>0.05),细胞迁移能力下降,差异有统计学意义(t=5.121,P<0.05),转化生长因子β(TGF-β)诱导的炎性细胞因子IL-6、MCP-1及IL-1β分泌能力下降,差异有统计学意义(t=9.371、5.690、6.192,P<0.05).结论 EMMPRIN c.889C>A突变不影响EMMPRIN蛋白表达及血管平滑肌细胞增殖,但可以抑制血管平滑肌细胞迁移及炎症细胞因子分泌.
Abstract
Objective To explore the effects of extracellular matrix metalloproteinase inducer(EMMPRIN)on the proliferation,migration and inflammatory response of vascular smooth muscle cells(VSMCs).Methods CRISPR/Cas9 technology was used to construct EMMPRIN c.889C>A-point mutation VSMC line.Real-time qPCR and Western blot were performed to detect the expression level of EMMPRIN.Cell proliferation activity was measured using the CCK-8 assay,while cell migration capability was evaluated using the Transwell assay.Secretion of interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1),and interleukin-1β(IL-1β)was detected using enzyme-linked immunosorbent assay(ELISA).Results Compared to the wild-type(WT)cells,the expression level of EMMPRIN did not exhibit significant changes in c.889C>A mutant cells(t=0.732,P>0.05).There was no statistically significant difference in cell proliferation activity(P>0.05),while cell migration capacity significantly decreased(t=5.121,P<0.05).Secretion of inflammatory cytokines IL-6,MCP-1 and IL-1β induced by TGF-β decreased markedly in the c.889C>A mutant cells(t=9.371,5.690,6.192;P<0.05).Conclusion The EMMPRIN c.889C>A mutation does not affect the expression of EMMPRIN or VSMCs proliferation.However,it can supress the migration of VSMCs and the secretion of inflammatory cytokines.
关键词
细胞外基质金属蛋白酶诱导子/血管平滑肌细胞/增殖/迁移Key words
Extracellular matrix metalloproteinase inducer/Vascular smooth muscle cells/Proliferation/Migration引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据