下肢动脉粥样硬化斑块内miRNA-199a-3p对外周血单核细胞的影响

Influence of miRNA-199a-3p on peripheral blood mononuclear cells in atherosclerotic plaques of lower extremities

目的分析miRNA-199a-3p对外周血单核细胞的影响。方法选取下肢动脉粥样硬化闭塞症患者(实验组)及健康体检者(对照组),各60例。取1例截肢患者截肢远端血管,以近端血管为对照,采用实时荧光定量聚合酶链式反应(PCR)检测,获得血管组织及人外周血中单核细胞和微泡(MV)中miRNA-199a-3p的表达水平,此外,于细胞对数生长期,分设miRNA-199a-3p inhibitor组、miRNA-199a-3p mimics组、空白对照组,各组设复孔6个,转染miRNA-199a-3p inhibitor、miRNA-199a-3p mimics、FuGENE,分析miRNA-199a-3p对外周血单核细胞的影响。结果动脉粥样硬化斑块内miRNA-199a-3p表达水平(8.49±3.05)高于正常组织的(3.64±1.20),差异有统计学意义(P<0.05)。实验组外周血单核细胞、MV中的miRNA-199a-3p表达水平高于对照组,差异有统计学意义(P<0.05)。miRNA-199a-3p inhibitor组的外周血单核细胞中的miRNA-199a-3p表达水平低于空白对照组,miRNA-199a-3p mimics组的外周血单核细胞中miRNA-199a-3p表达水平高于空白对照组,差异有统计学意义(P<0.05)。检测Fractalkine、T细胞表达和分泌因子(RANTES)、单核细胞趋化因子-1(MCP-1)的mRNA表达水平,miRNA-199a-3p inhibitor组高于空白对照组,miRNA-199a-3p mimics组低于空白对照组,差异有统计学意义(P<0.05)。结论下肢动脉粥样硬化斑块内miRNA-199a-3p通过MV作用于单核细胞,抑制其Fractalkine、RANTES、MCP-1等趋化因子表达,而起到抗炎抗动脉粥样硬化的作用。

Objective To analyze the influence of miRNA-199a-3p on peripheral blood mononuclear cells. Methods A total of 60 patients with arteriosclerosis obliterans of lower extremity were selected as experimental group, and 60 healthy persons were selected as control group. The distal vessels were taken from one amputated patient, and the proximal vessels were used as controls. Real time fluorescence quantitative polymerase chain reaction（PCR） was used to detect the blood vessels. The expression level of miRNA-199a-3p in vascular tissue and human peripheral blood monocytes and microvesicles（MV） were obtained. In addition, the miRNA-199a-3p inhibitor group, miRNA-199a-3p mimics group and blank control group were set up during the logarithmic growth phase of cells, 6 holes were set up in each group to transfect miRNA-199a-3p inhibitor, miRNA-199a-3p mimics and Fu GENE. The influence of miRNA-199a-3p on peripheral blood mononuclear cells was analyzed. Results The expression level of miRNA-199a-3p in atherosclerotic plaques of lower extremities was（8.49±3.05）, which was significantly higher than（3.64±1.20） in the normal tissues, the difference was statistically significant（P〈0.05）. In the experimental group, the expression level of miRNA-199a-3p in monocytes and MV on peripheral blood were significantly higher than those in the control group, the differences were statistically significant（P〈0.05）. The expression level of miRNA-199a-3p in peripheral blood monocytes of miRNA-199a-3p inhibitor group was significantly lower than that of the blank control group, the expression level of miRNA-199a-3p inperipheral blood monocytes of miRNA-199a-3p mimics group was significantly higher than that of the blank control group, the differences were statistically significant（P〈0.05）. The m RNA expression level of Fractalkine, regulated upon activation normal T-cell expressed and secreted（RANTES） and monocyte chemoattractant protein-1（MCP-1） were detected, miRNA-199a-3p inhibitor group were higher than those of blank control group, miRNA-199a-3p mimics group was lower than those of blank control group, the differences were statistically significant（P〈0.05）. Conclusion miRNA-199a-3p in the atherosclerotic plaques of the lower extremities affects monocytes through MV, inhibits the expression of chemokines such as Fractalkine, RANTES and MCP-1, and plays an important role in anti inflammation and anti atherosclerosis.