腺病毒介导心脏特异性过表达RIP140对大鼠心脏功能及炎症通路的影响

摘要

目的 探讨心脏组织特异性过表达受体相互作用蛋白140(receptor interacting protein 140,RIP140)对心脏功能及炎症通路的影响.方法 大鼠心脏组织多点注射携带RIP140基因的腺病毒载体,使心肌组织过表达RIP140;免疫荧光验证RIP140蛋白在心脏组织中的过表达情况;超声心动图与血流动力学参数评估心脏功能;ELISA分析TNF-α、IL-2、IL-1β等炎症因子水平;Western blot分析p65、IκB-α的蛋白水平.结果 腺病毒载体介导的外源基因RIP140可成功过表达于心脏组织,诱导心室扩张、左室射血分数下降、心功能受损,促进心肌组织TNF-α、IL-2、IL-1β炎症因子释放,p65蛋白入核、胞质IκB-α蛋白降解.结论 腺病毒介导RIP140基因在心脏组织中的特异性过表达损伤心脏功能,激活NF-κB/p65炎症通路,促进炎症因子释放.%Aim To explore the role of cardiac-specific overexpression of RIP140 in cardiac function and inflammation signaling pathway.Methods Direct intra-myocardial injection of adenovirus vector expressing RIP140 drove transgene expression in heart tissue.RIP140 overexpression was confirmed using immunofluorescence technique in heart.Cardiac function was assessed by echocardiographic and hemodynamic assessment.TNF-α,IL-2 and IL-1β inflammatory cytokines were detected by ELISA,and the protein levels of p65 and IκB-α were measured by Western blot.Results Adenovirus-mediated foreign gene of RIP140 was successfully transferred in cardiac tissue.RIP140 overexpression in heart induced ventricular dilation,decreased left ventricular ejection fraction and cardiac malfunction,as well as increased releases of TNF-α,IL-2 and IL-1β inflammatory cytokines,p65 protein translocation into the nucleus and IκB-α protein degradation in cytoplasm.Conclusion Adenovirus-mediated RIP140 overexpression in cardiac tissue impairs cardiac function,activates NF-κB/p65 inflammatory signaling pathway and induces the release of inflammatory cytokines.