miR-31通过下调FIH和上调VEGF-A/VEGFR-2促进大鼠主动脉内皮细胞增殖  

miR-31 promotes the proliferation of rat aortic endothelial cells by downregulating FIH and upregulating VEGF-A/VEGFR-2

作　　者：李咏梅 付媛 曹利利 杜若琛 王春芳 张轩萍 Li Yongmei;Fu Yuan;Cao Lili;Du Ruochen;Wang Chunfang;Zhang Xuanping(Key Laboratory of Cell Physiology,Ministry of Education,Department of Physiology,Shanxi Medical University,Taiyuan,shanxi 030001,China;Department of Pharmacology,School of Basic Medicine,Shanxi Medical University,Jinzhong,shanxi,030619,China;Laboratory Animal Center,Shanxi Medical University,Taiyuan,shanxi,030001,China)

机构地区：[1]山西医科大学生理学系细胞生理学教育部重点实验室,太原030001 [2]山西医科大学基础医学院药理学教研室,晋中030619 [3]山西医科大学实验动物中心,太原030001

出　　处：《中国组织化学与细胞化学杂志》2023年第4期339-346,共8页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金项目(82001326);中央引导地方科技发展项目(YDZJSX2022A056);山西省应用基础研究计划项目(201901D211319);山西省基础研究项目(20210302124172)。

摘　　要：目的探讨miR-31对大鼠主动脉内皮细胞(rat aortic endothelial cells,RAECs)增殖的影响及其潜在的作用机制。方法分离RAECs,鉴定为目的细胞后,观察miR-31激动剂(miR-31 agomir)和抑制剂(miR-31 antagomir)对RAECs增殖以及缺氧诱导因子抑制因子(hypoxia inducible factor inhibitor,FIH)、血管内皮生长因子A(vascular endothelial growth factor A,VEGF-A)和血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR-2)水平的影响。通过CCK-8实验测定miR-31对细胞增殖能力的影响,qRT-PCR检测细胞中miR-31的相对表达水平,Western blot检测细胞中FIH、VEGF-A和VEGFR-2水平。结果在无特殊器械、不添加营养因子的情况下成功培养出RAECs;转染miR-31 agomir显著促进RAEC的细胞增殖,降低FIH水平,上调VEGF-A和VEGFR-2水平,转染miR-31 antagomir则相反。结论miR-31可能通过下调FIH、上调VEGF-A和VEGFR-2的表达促进主动脉内皮细胞增殖。Objective To investigate the impact of miR-31 on the proliferation of rat aortic endothelial cells(RAECs)and its potential mechanisms,laying a foundation for exploring endothelial cell functional characteristics and related cardiovascular disease research.Methods After isolating RAECs and identifying them as target cells,the effects of miR-31 agonist(miR-31 agomir)and inhibitor(miR-31 antagomir)on the proliferation of RAECs and on the levels of hypoxia-inducible factor inhibitor(FIH),vascular endothelial growth factor A(VEGF-A),and vascular endothelial growth factor receptor 2(VEGFR-2)in RAECs were observed.The impact of miR-31 on cell proliferation was measured using the CCK-8 assay,the relative expression level of miR-31 in cells was detected by qRT-PCR,and the levels of FIH,VEGF-A,and VEGFR-2 in cells were assessed by Western blot.Results RAECs were successfully cultured without special equipment or adding nutritional factors;transfection with miR-31 agomir significantly promoted RAEC proliferation,decreased FIH levels,and upregulated VEGF-A and VEGFR-2 levels,while transfection with miR-31 antagomir had the opposite effect.Conclusion miR-31 may promote the proliferation of aortic endothelial cells by downregulating FIH and upregulating the expression of VEGF-A and VEGFR-2.

关 键 词：MiR-31 主动脉内皮细胞 原代培养 VEGF 增殖 

分 类 号：R541.9[医药卫生—心血管疾病]