间充质干细胞来源外泌体通过microRNA-210发挥对H2O2诱导的心肌损伤的保护作用  

Mechanism of microRNA-210 on myocardium repair regulated by mesenchymal stem cell exosomes.

作　　者：郝媛媛 马爱群[2] 吴冠吉[1] 李涛[1] 胡建华 HAO Yuan-yuan;MA Ai-qun;WU Guan-ji(Department of Cardiology,Xi'an Central Hospital,Xi'an Shaanxi 710032,China;Department of Cardiology,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an Shaanxi 710032,China)

机构地区：[1]西安市中心医院心血管内科,陕西西安710032 [2]西安交通大学第一附属医院心血管内科,陕西西安710032

出　　处：《临床和实验医学杂志》2020年第24期2577-2581,共5页Journal of Clinical and Experimental Medicine

基　　金：国家科技部重点项目(编号:2017YFC1308302);西安市卫生健康委员会项目(编号:2020yb01)。

摘　　要：目的研究间充质干细胞(MSC)外泌体经通过microRNA-210发挥H2O2诱导的心肌损伤的保护作用及其机制。方法应用H2O2溶液诱导建立H9c2心肌细胞的氧化应激损伤模型,采用噻唑蓝(MTT)检测其对H9c2细胞生存率的影响。应用实时定量PCR检测不同浓度H2O2溶液对MSC外泌体中microRNA-210表达水平的影响。应用慢病毒感染建立microRNA-210 precursor、microRNA-210 inhibitor组,检测microRNA-210对H2O2诱导的H9c2心肌细胞存活率的影响。分别采用0.5 mmol/L的H2O2处理和不用H2O2处理处理microRNA-210 precursor组、microRNA-210 inhibitor组和H9c2对照组细胞24 h,再应用聚合酶链式反应(PCR)法检测各组细胞中的CASP8AP2、caspase-8和caspase-3基因的mRNA的相对表达。结果H2O2可降低H9c2心肌细胞的生存率并诱导MSC外泌体中microRNA-210表达水平增加,其作用强度均呈浓度依赖性(P<0.05)。microRNA-210 precursor组的H9c2心肌细胞生存率[(0.66±0.16)%]较未转染组H9c2细胞对照组[(0.41±0.10)%]显著增加(P<0.01),microRNA-210 inhibitor组的生存率[(0.29±0.07)%]较H9c2细胞对照组显著降低(P<0.001)。与H9c2细胞对照组(2.93±0.56、2.22±0.27、2.10±0.36)相比,microRNA-210-precursor组CASP8AP2、caspase-8和caspase-3 mRNA的相对表达(1.01±0.13、1.00±0.11、1.01±0.12)均显著降低,H9c2细胞+H2O2组以上指标[(7.22±1.32、6.10±1.03、5.98±0.44)]均表达均显著增加,H9c2细胞+H2O2组也较microRNA-210-precursor组显著升高(P<0.05);与H9c2+H2O2组相比,microRNA-210-precursor+H2O2组(3.09±0.60、2.25±0.29、2.12±0.31)CASP8AP2、caspase-8和caspase-3 mRNA的相对表达均显著降低(P<0.05)。结论H2O2可诱导H9c2心肌细胞损伤,并促进外泌体中microRNA-210表达的增加,而过表达microRNA-210可抑制H2O2诱导的心肌细胞的损伤,因此microRNA-210发挥抗心肌细胞损伤作用的可能机制是通过抑制CASP8AP2通路相关基因的表达而实现的。Objective To investigate the mechanism of microRNA-210 on myocardium repair regulated by mesenchymal stem cells(MSC)exosomes.Methods The oxidative stress model of rats H9c2 myocardial cells were induced by hydrogen peroxide(H2O2)and the survival rate was detected by MTT method.Quantitative real-time PCR was conducted to detect expression of microRNA-210 in MSC exosomes under H2O2 treatment.The high-and low-expression model of microRNA-210 were constructed via lentivirus infection of H9c2 cells,and cell viability was detected by MTT to investigate the effect of microRNA-210 on H9c2 cells under H2O2 treatment.Cells in the microRNA-210 precursor group,microRNA-210 inhibitor group and H9c2 control group were treated with 0.5 mmol/L H2O2 and without H2O2 treatment for 24 hours,and then polymerase chain reaction(PCR)method was used to detect the concentration of cells in each group.The relative expression of mRNA of CASP8AP2,caspase-8 and caspase-3 genes.Results H2O2 can reduce the survival rate of H9c2 cardiomyocytes and induce an increase in the expression level of microRNA-210 in MSC exosomes,and the intensity of which is concentration-dependent(P<0.05).The survival rate of H9c2 cardiomyocytes in the microRNA-210 precursor group(0.66±0.16)%was significantly higher than that of the untransfected H9c2 cell control group(0.41±0.10)%(P<0.01),and the survival rate of the microRNA-210 inhibitor group(0.29±0.07)%was significantly lower than that of the H9c2 cell control group(P<0.001).Compared with the H9c2 cell control group(2.93±0.56,2.22±0.27,2.10±0.36),the relative expressions of CASP8AP2,caspase-8 and caspase-3 mRNA in the microRNA-210-precursor group(1.01±0.13,1.00±0.11,1.01±0.12)were significantly reduced,and the expression of the above indicators in the H9c2 cell+H2O2 group were significantly increased,and the H9c2 cell+H2O2 group(7.22±1.32,6.10±1.03,5.98±0.44)were also significantly higher than that in the microRNA-210-precursor group(P<0.05);Compared with the H9c2+H2O2 group,the mRNA relative expressi

关 键 词：间充质干细胞 外泌体 microRNA-210 心肌细胞损伤 CASP8AP2通路 

分 类 号：R-332[医药卫生] R542.2