敲低miR-34c-5p抑制低氧诱导的大鼠骨髓间充质干细胞的凋亡  

miR-34c-5p knockdown inhibits hypoxia induced cell apoptosis of bone marrow derived mesenchymal stem cells in rats

作　　者：董扬[1] 张芬[1] 李幸幸 吴杰 徐忠诚 DONG Yang;ZHANG Fen;LI Xing-xing;WU Jie;XU Zhong-cheng(The Second Department of Cardiology,Jinhua People's Hospital,Jinhua 321099,China)

机构地区：[1]金华市人民医院心内二科,浙江金华321099

出　　处：《基础医学与临床》2022年第11期1690-1696,共7页Basic and Clinical Medicine

基　　金：金华市科技计划项目(2019-3-021)。

摘　　要：目的探讨敲低miR-34c-5p对提高骨髓间充质干细胞(BM-MSCs)存活率的作用。方法将大鼠BM-MSCs分为常氧组(normoxia组)、低氧组(hypoxia组)、常氧抑制剂阴性对照组(normoxia inhibitor NC组)、低氧抑制剂阴性对照组(hypoxia inhibitor NC组)、常氧微RNA(miR)抑制剂组(normoxia miR inhibitor组)和低氧miR抑制剂组(hypoxia inhibitor miR组);Lipofectamine2000将miR-34c-5p inhibitor转染至BM-MSCs细胞;实时荧光定量PCR(RT-qPCR)和ELISA检测miR-34c-5p、IGF、HGF、bFGF和VEGF表达;流式细胞测量术和TUNEL检测细胞凋亡,Western blot检测cleaved caspase-3,-9,Bcl-2和Bax蛋白水平;生化试剂盒检测ATP/ADP。流式细胞测量术检测细胞摄取葡萄糖能力。结果与hypoxia inhibitor NC组相比,敲低miR-34c-5p可明显抑制MSCs凋亡(P<0.01),显著降低cleaved caspase-3,-9和Bax蛋白表达,提高Bcl-2蛋白表达(P<0.01);IGF、HGF、bFGF和VEGF表达显著升高(P<0.05,P<0.01);ATP/ADP比率和细胞摄取葡萄糖的能力显著提高(P<0.05,P<0.01)。结论敲低miR-34c-5p可抑制低氧诱导的大鼠BM-MSCs的凋亡。Objective To explore the effect of miR-34 c-5 p knockdown on improving the survival rate of bone marrow derived mesenchymal stem cells(BM-MSCs).Methods The BM-MSCs were divided into normoxia group,hypoxia group,normoxia inhibitor NC group,hypoxia inhibitor NC group,normoxia microRNA(miR)inhibitor group and hypoxia miR inhibitor group.The expression level of miR-34 c-5 p,IGF,HGF,bFGF and VEGF was detected by real-time quantitative PCR(RT-qPCR)and ELISA.Cell apoptosis was detected by flow cytometry and TUNEL.The protein expression levels of cleaved caspase-3,-9,Bcl-2 and Bax were detected by Western blot.ATP/ADP was detected by biochemical kits,and glucose uptake was detected by flow cytometry.Results Compared with hypoxic inhibitor NC group,miR-34 c-5 p knockdown significantly inhibited BM-MSCs apoptosis(P<0.01),reduced protein expression of cleaved caspase-3,-9 and Bax and increased protein expression by Bcl-2(P<0.01).The expression of IGF,HGF,bFGF and VEGF(P<0.05,P<0.01)were significantly increased;ATP/ADP rate and cell capacity of glucose transporting also increased(P<0.05,P<0.01).Conclusions miR-34 c-5 p knockdown may antagonize hypoxia induced cell apoptosis of BM-MSCs in rat.

关 键 词：敲低miR-34c-5p 骨髓间充质干细胞 低氧 细胞凋亡 

分 类 号：R965.2[医药卫生—药理学]