miR-27b-3p对主动脉血管平滑肌细胞和单核细胞功能的影响及机制研究  

Effect of miR-27b-3p on the function of aortic vascular smooth muscle cells and monocytes and the mechanism

作　　者：肖远扬 李良学 XIAO Yuanyang;LI Liangxue(Department of Vascular Surgery,Puren Hospital Affiliated to Wuhan University of Science and Technology,Wuhan 430081,China)

机构地区：[1]武汉科技大学附属普仁医院血管外科,湖北武汉430081

出　　处：《中国普通外科杂志》2022年第12期1644-1652,共9页China Journal of General Surgery

基　　金：湖北省武汉市卫健委课题基金资助项目(WX21D52)。

摘　　要：背景与目的:大数据基因芯片数据分析发现,在腹主动脉瘤(AAA)组织与患者血清中miR-27b-3p表达显著上调。然而,miR-27b-3p在主动脉血管平滑肌细胞(VSMC)中的功能及作用机制尚不清楚。因此,本研究探讨miR-27b-3p在VSMC中的功能,及其对单核巨噬细胞、细胞外基质的影响和作用机制。方法:将VSMC转染miR-27b-3p抑制物后,用CCK-8检测细胞增殖活力,流式细胞术检测细胞凋亡。将单核巨噬细胞THP-1转染miR-27b-3p抑制物后,用ELISA法检测炎症因子表达,Western blot检测基质相关蛋白表达水平。用双萤光素酶报告基因实验以及相关功能实验验证miR-27b-3p和PTEN的靶向关系。结果:miR-27b-3p敲低后,VSMC增殖能力明显增强,细胞凋亡率明显降低(均P<0.05)。miR-27b-3p敲低后,THP-1中促炎症因子(TNF-α、IL-12)表达水平、基质金属蛋白酶9 (MMP-9)表达水平明显降低,而抑炎因子(IL-4、IL-10)表达水平、金属蛋白酶抑制剂-1 (TIMP-1)表达水平明显升高(均P<0.05)。双萤光素酶实验证实miR-27b-3p的靶基因为PTEN。miR-27b-3p过表达的VSMC中PTEN蛋白表达水平明显下调(P<0.05),而PTEN过表达的VSMC增殖活力明显增强、凋亡比例明显减少(P<0.05)。PTEN过表达THP-1的TNF-α、IL-12表达水平与MMP-9蛋白表达水平下调,同时IL-4、IL-10浓度与TIMP-1蛋白表达水平上调(均P<0.05)。miR-27b-3p和PTEN同时过表达后,VSMC与THP-1以上指标的变化与对照组差异均无统计学意义(均P>0.05)。结论:miR-27b-3p可通过靶向PTEN参与调节VSMC增殖和凋亡,抑制单核巨噬细胞的炎症反应和细胞外基质蛋白表达。miR-27b-3p/TPEN分子轴可能在AAA发生和进展过程发挥重要作用。Background and Aims: Big data gene chip analysis showed that the expression of miR-27b-3p is significantly up-regulated in abdominal aortic aneurysm(AAA) tissue and patients’ serum. However, the function and action mechanism of miR-27b-3p in aortic vascular smooth muscle cells(VSMCs) remains unclear. Therefore, this study was conducted to investigate the function of miR-27b-3p in VSMCs, as well as its effects on macrophages and the extracellular matrix, and the action mechanism.Methods: In VSMCs after transfection with miR-27b-3p inhibitor, the cell proliferation activity was detected by CCK-8, and the apoptosis was detected by flow cytometry. In mononuclear macrophage cell line, THP-1 cells after transfection with miR-27b-3p inhibitor, the expressions of inflammatoryassociated factors were detected by ELISA, and the expression levels of matrix-related proteins were detected by Western blot. The targeting relationship between miR-27b-3p and PTEN was verified by dual luciferase reporter gene assay and related functional experiments.Results: In VSMCs after miR-27b-3p knockdown, the proliferative ability was significantly increased, while the apoptotic rate was significantly decreased(both P<0.05). In THP-1 cells after miR-27b-3p knockdown, the expression levels of pro-inflammatory factors(TNF-α, IL-12) and the matrix metalloproteinase 9(MMP-9) were significantly decreased, while the expression levels of inflammatory factors(IL-4, IL-10) and metalloproteinase inhibitor 1(TIMP-1) were significantly increased(all P<0.05). Dual-luciferase experiments confirmed that PTEN was a target gene of miR-27b-3p. The expression level of PTEN protein was significantly down-regulated in VSMCs with miR-27b-3p overexpression, while the proliferation was significantly enhanced and the apoptotic rate was significantly decreased in VSMCs with PTEN overexpression(all P<0.05). The expression levels of TNF-α and IL-12 as well as MMP-9 protein were significantly down-regulated, while the levels of IL-4 and IL-10 as well as TIMP-1 protei

关 键 词：主动脉瘤 腹 肌 平滑 血管 单核细胞 炎症反应 细胞外基质 微RNAS 

分 类 号：R654.3[医药卫生—外科学]