Hippo信号通路参与心肌缺血后适应的保护作用  

Involvement of Hippo pathway in cardioprotection of ischemic postconditioning

作　　者：周露[1] 唐广胜 朱竑翊 ZHOU Lu;TANG Guangsheng;ZHU Hongyi(Kangda College of Nanjing Medical University,Lianyungang 222000,China)

机构地区：[1]南京医科大学康达学院,江苏连云港222000 [2]南京医科大学,南京210029

出　　处：《实用医学杂志》2018年第18期3000-3005,共6页The Journal of Practical Medicine

基　　金：南京医科大学康达学院科研发展基金重点项目(编号:KD2016KYJJZD004)

摘　　要：目的研究Hippo信号通路是否参与心肌缺血后适应及其参与机制。方法将转染了MST1-siRNA和阴性对照NC-siRNA的乳鼠心肌细胞分别建立缺氧/复氧(H/R)和缺氧后适应(HPostC)模型;采用流式细胞术检测细胞凋亡率,MTT比色法检测细胞活性,碧云天MDA试剂盒检测细胞MDA含量,最后比较转染MST1-siRNA细胞与转染NC-siRNA细胞中后适应改善程度的差异,以(HPostC-H/R)/H/R的值表示;使用western blot检测MST1和YAP的蛋白表达及磷酸化水平;利用免疫共沉淀法检测Bcl-xL与MST1和Bax之间的相互作用。结果转染MST1-siRNA细胞与转染NC-siRNA细胞相比,后适应组抗细胞凋亡的作用减弱[(54.96±1.52)%vs.(82.13±3.02)%,P <0.05],增强细胞活性的作用减弱[(22.32±4.42)%vs.(61.53±6.69)%,P <0.05],减少脂质氧化产物MDA的作用减弱[(32.06±6.44)%vs.(53.11±9.43)%,P <0.05];与H/R相比,HPostC降低MST和YAP磷酸化水平,削弱MST1与Bcl-xL之间的相互作用,抑制Bcl-xL与Bax的解离(P <0.05)。结论 Hippo信号通路参与缺血后适应保护作用,其机制是降低YAP磷酸化水平,并且削弱MST1与Bcl-xL之间的相互作用,抑制Bcl-xL与Bax解离,从而发挥抗细胞凋亡作用。Objective To explore the role of the Hippo pathway in cardioprotection of ischemic postconditioning and the mechanism.Methods Specific siRNA targeting MST1 gene(MST1-siRNA)or negative controlsiRNA(NC-siRNA)was transfected into neonatal rat cardiomyocytes by liposome.Then cardiomyocytes were subjected to hypoxia/reoxygenation(H/R)or hypoxia postconditioning(HPostC)with a hypoxia tank.Cell apoptosis and cell viability were measured respectively by Annexin V-FITC staining with a flow-cytometer and MTT colorimetric assay.MDA levels were determined by MDA assay.The value of(HPost-CH/R)/H/R indicated the degree action of HPostC.Western blot was used to measure the expressions and phosphorylation of MST1 and YAP.COIP was used to determine the association between BclxL and either MST1 or Bax.Results The role of HPostC played in anticell apoptosis[(54.96±1.52)%vs.(82.13±3.02)%,P<0.05],enhancing cell viability[(22.32±4.42)%vs.(61.53±6.69)%,P<0.05]and decreasing MDA level[(32.06±6.44)%vs.(53.11±9.43)%,P<0.05]in MST1-siRNA transfected cells were all attenuated as compared with those in NC-siRNA transfected cells.In addition,as compared with H/R group,HPostC significantly decreased phosphorylation levels of MST1 and YAP and weakened the interaction of MST1 and BclxL,and enhanced the association between Bcl-xL and Bax.Conclusions Hippo pathway is involved in cardioprotection of ischemic postconditioning through decreasing phosphorylation of YAP and weakening the interaction of MST1 and Bcl?xL.

关 键 词：缺血后适应 Hippo信号通路 MST YAP 

分 类 号：R54[医药卫生—心血管疾病]