下调miR-106a-5p对过氧化氢诱导的心肌细胞损伤及JAK1/STAT3通路的影响  

The effects of down-regulation of miR-106a-5p on hydrogen peroxide-induced cardiomyocyte damage and JAK1/STAT3 pathway

作　　者：姜花[1] 沈延梅[1] 马驯凯 JIANG Hua;SHEN Yanmei;MA Xunkai(Department of Cardiology,Qinghai Red Cross Hospital,Xining 810001,China)

机构地区：[1]青海红十字医院心内科,810001

出　　处：《天津医药》2023年第2期113-117,共5页Tianjin Medical Journal

基　　金：青海省重点研发与转化计划项目(175110013106445)。

摘　　要：目的 探讨下调miR-106a-5p对过氧化氢(H2O2)诱导的心肌细胞损伤及Janus激酶1(JAK1)/信号转导和转录激活因子3(STAT3)通路的影响。方法 不同浓度梯度H2O2(0、50、100、200、400μmol/L)处理大鼠心肌细胞H9c2。H9c2细胞分为对照组、H2O2组(100μmol/L H_(2)O_(2))、miR-106a-5p NC组(100μmol/L H_(2)O_(2)+转染miR-106a-5p NC)和miR-106a-5p siRNA组(100μmol/L H_(2)O_(2)+转染miR-106a-5p siRNA)。实时荧光定量PCR(qPCR)法检测miR-106a-5p表达;CCK-8法检测细胞增殖;流式细胞仪检测细胞凋亡;试剂盒检测细胞培养液中乳酸脱氢酶(LDH)含量、谷胱甘肽过氧化物酶(GSH-Px)活性及细胞中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;蛋白免疫印迹法检测细胞中JAK1/STAT3通路相关蛋白表达。结果 根据H_(2)O_(2)浓度梯度实验,选择100μmol/L H_(2)O_(2)用于后续实验。与对照组相比,H2O2组H9c2细胞增殖抑制率、凋亡率、细胞中miR-106a-5p水平、MDA含量、p-JAK1/JAK1、p-STAT3/STAT3及细胞培养液中LDH含量显著升高(P<0.05),细胞培养液GSH-Px活性及细胞中SOD活性显著降低(P<0.05);上述指标在H2O2组与miR-106a-5p NC组间的差异无统计学意义(P>0.05);与miR-106a-5p NC组相比,miR-106a-5p siRNA组H9c2细胞增殖抑制率、凋亡率、细胞中miR-106a-5p水平、MDA含量、p-JAK1/JAK1、p-STAT3/STAT3及细胞培养液中LDH含量显著降低(P<0.05),细胞培养液GSH-Px活性及细胞中SOD活性显著升高(P<0.05)。结论 下调miR-106a-5p可抑制JAK1/STAT3通路激活,提高抗氧化应激水平,减轻H_(2)O_(2)诱导的心肌细胞损伤。Objective To investigate the effects of down-regulation of miR-106a-5p on hydrogen peroxide(H2O2)-induced cardiomyocyte damage and Janus kinase 1(JAK1)/signal transducer and activator of transcription 3(STAT3)pathway.Methods Different concentration gradients of H2O2(0,50,100,200 and 400 μmol/LH_(2)O_(2)) were used to treat rat cardiomyocytes H9c2.Rat cardiomyocytes H9c2 were divided into the control group,the H_(2)O_(2)group(100 μmol/L H_(2)O_(2)),the miR-106a-5p NC group(100 μmol/L H_(2)O_(2)+transfected with miR-106a-5p NC) and the miR-106a-5p siRNA group(100 μmol/L H_(2)O_(2)+ transfected with miR-106a-5p siRNA).Real-time fluorescent quantitative PCR(qPCR) method was used to detect the expression of miR-106a-5p.CCK-8 method was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.LDH content,GSH-Px activity in cell culture fluid,MDA content and SOD activity in cells were detected.Western blotting(WB) was used to detect the expression of JAK1/STAT3 pathway related proteins in cells.Results According to the H_(2)O_(2)concentration gradient experiment,100 μmol/L H_(2)O_(2) was selected for subsequent experiments.Compared with the control group,the proliferation inhibition rate,apoptosis rate,miR-106a-5p level,MDA content,p-JAK1/JAK1,p-STAT3/STAT3 protein levels in H9c2 cells and LDH content in cell culture fluid were significantly increased in the H2O2group(P <0.05).The GSH-Px activity in cell culture fluid and SOD activity in cells significantly reduced(P <0.05).There were no significant differences in above indicators between the H_(2)O_(2)group and the miR-106a-5p NC group(P> 0.05).Compared with the miR-106a-5p NC group,the proliferation inhibition rate,apoptosis rate,miR-106a-5p level,MDA content,p-JAK1/JAK1,p-STAT3/STAT3 protein levels in H9c2 cells and LDH content in cell culture fluid were reduced significantly in the miR-106a-5p siRNA group(P<0.05),and the GSH-Px activity in cell culture fluid and SOD activity in cells increased significantly(P <0.05).Conclusion Down-reg

关 键 词：肌细胞 心脏 过氧化氢 微RNAS Janus激酶1 STAT3转录因子 氧化性应激 

分 类 号：R349.5[医药卫生—基础医学]