miR-129-5p调控PGC-1α/SIRT3信号通路对肺动脉平滑肌细胞活性及VEGF水平的影响  

The effect of miR-129-5p regulating PGC-1α/SIRT3 signaling pathway on the activity and VEGF level of pulmonary artery smooth muscle cells

作　　者：魏士刚 马雪梅 宿海峰 WEI Shi-Gang;MA Xue-Mei;SU Hai-Feng(Beijing Aerospace General Hospital,Beijing 100076,China)

机构地区：[1]北京航天总医院,北京100076 [2]佳木斯大学附属第一医院

出　　处：《中国老年学杂志》2024年第14期3469-3474,共6页Chinese Journal of Gerontology

基　　金：黑龙江省卫生计生委科研项目(No.2017-399)。

摘　　要：目的 探究miR-129-5p调控过氧化物酶体增殖物激活受体γ辅助激活因子(PGC)-1α/沉默信息调节因子(SIRT)3信号通路对肺动脉平滑肌细胞(PASMC)活性及血管内皮生长因子(VEGF)水平的影响。方法 将人原代PASMC(HPASMC)按照随机数字表法分为:缺氧组(缺氧环境中培养的HPASMC不转染任何质粒)、miR-129-5p组(缺氧环境中培养的HPASMC转染miR-129-5p mimic质粒)、miR-NC组(缺氧环境中培养的HPASMC转染mimic NC质粒)、si-PGC-1α组(缺氧环境中培养的HPASMC同时转染si-PGC-1α和miR-129-5p mimic质粒)。RT-聚合酶链反应(PCR)检测常氧和缺氧HPASMC中miR-129-5p表达;CCK-8检测细胞增殖;Transwell法检测细胞侵袭;流式细胞仪检测细胞凋亡;Western印迹检测细胞中PGC-1α、SIRT3和VEGF蛋白表达;荧光素酶报告检测PGC-1α和miR-129-5p的靶向关系。结果 和常氧HPASMC相比,缺氧HPASMC中miR-129-5p表达明显降低(P<0.05)。与缺氧组比较,miR-129-5p组细胞增殖明显降低(P<0.05)。与缺氧组及miR-NC组比较,miR-129-5p组侵袭数目及VEGF蛋白表达降低,凋亡率、PGC-1α、SIRT3蛋白表达升高,差异有统计学意义(P<0.05)。3′非翻译区(UTR)与miR-129-5p有结合点位。转染野生型PGC-1α(PGC-1α-WT)时,miR-129-5p组荧光素酶活性比miR-NC组明显低(P<0.05)。和miR-129-5p组相比,si-PGC-1α组增殖、侵袭能力、VEGF蛋白表达明显升高,细胞凋亡率、PGC-1α、SIRT蛋白表达明显降低(P<0.000 1)。结论 过表达miR-129-5p可抑制缺氧诱导的PASMC增殖、侵袭并诱导凋亡,减少VEGF表达,其作用机制可能和激活PGC-1α/SIRT3通路有关。Objective To explore the effects of miR-129-5p regulation of peroxisome proliferator activated receptor-γcoactiva-tor(PGC)-1α/silencing regulatory protein(SIRT)3 signaling pathway on the activity of pulmonary artery smooth muscle cells(PASMCs)and vascular endothelial growth factor(VEGF)levels.Methods Human primary PASMC(HPASMC)was randomly di-vided into hypoxia group(HPASMC cultured in hypoxic environment without transfecting any plasmids),miR-129-5p group(HPASMC cultured in hypoxic environment transfected with miR-129-5p mimic plasmid),miR-NC group(HPASMC transfected mim-ic NC plasmid cultured in hypoxic environment),si-PGC-1αgroup(HPASMC cultured in hypoxic environments transfected with si-PGC-1αand miR-129-5p mimic plasmid simultaneously).RT-polymerase chain reaction(PCR)was used to detect the expression of miR-129-5p in normoxic and hypoxic HPASMCs;CCK-8 was used to detect cell proliferation;Transwell method was used to detect cell invasion;cell apoptosis was detected by flow cytometry;Western blot was used to detect the expressions of PGC-1α,SIRT3 and VEGF protein in cells;fluorescence enzyme report was used to detect the relationship between PGC-1αtargeting and miR-129-5p.Results Compared with normoxic HPASMC,the expression of miR-129-5p in hypoxic HPASMC was significantly reduced(P<0.05).Com-pared with the hypoxia group,cell proliferation in miR-129-5p group was significantly decreased(P<0.05).Compared with hypoxia group and miR-NC group,the invasion number and VEGF protein expression were decreased,while the apoptosis rate,PGC-1αand SIRT3 protein expressions were increased in miR-129-5P group,the differences were statistically significant(P<0.05).3'untranslated region(UTR)had binding sites with miR-129-5p.After transfection with wild type PGC-1α(PGC-1α-WT),the luciferase activity in the miR-129-5p group was significantly lower than that in the miR-NC group(P<0.05).Compared with miR-129-5p group,the prolif-eration,invasion ability and VEGF protein expression were significantly increased,while the

关 键 词：miR-129-5p 过氧化物酶体增殖物激活受体γ辅助激活因子(PGC)-1α/沉默信息调节因子(SIRT)3通路 肺动脉平滑肌细胞 增殖 侵袭 凋亡 血管内皮生长因子(VEGF) 

分 类 号：R734[医药卫生—肿瘤]