文章摘要
梁 戎,李兰英,张国珍,苏布道,雷文秀.白藜芦醇通过调节miR⁃21⁃5p/PDCD4通路抑制血管平滑肌细胞增殖与迁移[J].南京医科大学学报,2020,(1):26~31
白藜芦醇通过调节miR⁃21⁃5p/PDCD4通路抑制血管平滑肌细胞增殖与迁移
Resveratrol inhibits proliferation and migration of vascular smooth muscle cells by regulating miR⁃21⁃5p/PDCD4 pathway
投稿时间:2019-06-10  
DOI:10.7655/NYDXBNS20200106
中文关键词: 高血压  血管平滑肌细胞  白藜芦醇  微小RNA⁃21⁃5p  程序性细胞死亡因子4
英文关键词: hypertension  vascular smooth muscle cell  resveratrol  microRNA⁃21⁃5p  progrmammed cell death 4
基金项目:内蒙古自治区自然科学基金[2017MS(LH)0851]
作者单位
梁 戎 内蒙古自治区人民医院老年医学中心内蒙古 呼和浩特 010020 
李兰英 内蒙古自治区人民医院老年医学中心内蒙古 呼和浩特 010020 
张国珍 内蒙古自治区人民医院老年医学中心内蒙古 呼和浩特 010020 
苏布道 内蒙古自治区人民医院老年医学中心内蒙古 呼和浩特 010020 
雷文秀 内蒙古自治区人民医院老年医学中心内蒙古 呼和浩特 010020 
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中文摘要:
      目的:探讨白藜芦醇治疗高血压的作用靶点及相关作用机制。方法:利用血管紧张素Ⅱ诱导大鼠胸大动脉平滑肌细胞A7r5模拟高血压时平滑肌细胞增殖细胞模型,并使用白藜芦醇与miR?21?5p mimic进行干预。利用CCK?8实验检测A7r5细胞活力;利用细胞划痕实验检测A7r5细胞的迁移能力;利用real?time PCR技术检测miR?21?5p的表达;利用Western blot技术检测细胞增殖相关蛋白PCNA、Cyclin D1、CDK4与细胞迁移相关蛋白MMP2、MMP9、PDCD4的表达。此外,选取对数生长期293T细胞,利用双荧光素酶报告基因检测技术验证miR?21?5p与PDCD4的靶向关系。结果:与正常对照组相比,模型组细胞活力、迁移能力及增殖和迁移相关蛋白PCNA、Cyclin D1、CDK4、MMP2、MMP9与miR?21?5p的表达水平显著上调,PDCD4蛋白的表达水平显著下降,且差异具有统计学意义(P < 0.01);与模型组相比,白藜芦醇处理组的细胞活力、迁移能力及增殖和迁移相关蛋白PCNA、Cyclin D1、CDK4、MMP2、MMP9与miR?21?5p的表达水平显著下调,PDCD4蛋白的表达水平显著上调,且差异具有统计学意义(P<0.001);而miR?21?5p mimic转染后细胞活力、迁移能力及增殖和迁移相关蛋白PCNA、Cyclin D1、CDK4、MMP2、MMP9与miR?21?5p的表达水平显著上调,PDCD4蛋白的表达水平显著下降且差异具有统计学意义(P<0.001),双荧光素酶检测显示miR?21?5p可显著抑制野生型PDCD4 3'UTR报告基因载体的荧光素酶活性(P < 0.01)。结论:白藜芦醇通过调节miR?21?5p/PDCD4通路,抑制血管平滑肌细胞增殖与迁移,进而对高血压起到一定的治疗作用。
英文摘要:
      Objective:To explore the target and related mechanism of resveratrol in the treatment of hypertension. Methods:Using angiotensin Ⅱ to induce rat aortic smooth muscle cells A7r5 to establish a hypertensive cell model and interventing with resveratrol and miR?21?5p mimic on this basis. CCK?8 assay was used to detect cell viability of A7r5 cells,wounding healing assay was used to detect migration ability of A7r5 cells,Real?time PCR was used to detect expression of miR?21?5p,Western blot was used to detect the expression of cell proliferation?related proteins PCNA,Cyclin D1 and CDK4,and cell migration associated proteins MMP2,MMP9 and PDCD4. In addition,the targeting relationship between miR?21?5p and PDCD4 was verified by dual luciferase reporter gene detection technology in logarithmic growth phase 293T cells. Results:Compared with the NC group,the cell viability,migration ability and the expression level of proliferation and migration?related proteins PCNA,Cyclin D1,CDK4,MMP2,MMP9 and miR?21?5p were significantly up?regulated,and the expression level of PDCD4 protein was significantly decreased in the NC+ model group,and the difference was statistically significant(P < 0.01);Compared with the NC+ model group,the cell viability,migration ability,and the expression level of proliferation and migration?related proteins PCNA,Cyclin D1,CDK4,MMP2,MMP9 and miR?21?5p were significantly down?regulated,and the expression level of PDCD4 protein was significantly up?regulated in the NC+ model+resveratrol group,and the difference was statistically significant(P < 0.01);Compared with the NC+ model+resveratrol group,cell viability,migration ability and the expression level of proliferation and migration related proteins PCNA,Cyclin D1,CDK4,MMP2,MMP9 and miR?21?5p was significantly up?regulated,and the expression level of PDCD4 protein was significantly decreased in the miR?21?5p mimic+model+resveratrol group,and the difference was statistically significant(P < 0.01). The dual luciferase assay miR?21?5p significantly inhibited the luciferase activity of the wild?type PDCD4 3’UTR reporter vector(P < 0.01). Conclusion:Resveratrol inhibits the proliferation and migration of vascular smooth muscle cells by regulating the miR?21?5p/PDCD4 pathway,which in turn has a certain therapeutic effect on hypertension.
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