piR-16744对缺氧—复氧诱导的小鼠原代心肌细胞焦亡作用及机制研究  

The Effect and Mechanism of piR-16744 on Hypoxia-reoxygenation-induced Pyroptosis of Primary Neonatal Mouse Cardiomyocytes

作　　者：王飞[1] 陈鑫哲 王凯[1] 单婵 WANG Fei;CHEN Xin-zhe;WANG Kai;SHAN Chan(Institute of Translational Medicine,Qingdao University, Qingdao 266021, China)

机构地区：[1]青岛大学转化医学研究院,青岛266021

出　　处：《青岛大学学报（自然科学版）》2022年第2期5-11,共7页Journal of Qingdao University(Natural Science Edition)

基　　金：国家自然科学基金青年项目(批准号:21708021)资助。

摘　　要：为探究piR-16744对H/R诱导的NMCs焦亡作用及机制,体外构建NMCs的H/R模型,设置缺氧6 h、12 h、24 h复氧6 h的时间梯度。使用RT-qPCR检测piR-16744的表达水平,WB检测焦亡相关蛋白GSDMD、GSDMD-C、Caspase-1及炎症因子IL-1β、IL-18的表达水平,LDH和PI染色检测细胞死亡率。结果显示,H/R 6/6 h时,NMCs中焦亡相关蛋白的表达水平最高。抑制NMCs中piR-16744的表达可促进焦亡相关蛋白表达,细胞死亡率升高;过表达piR-16744则抑制焦亡相关蛋白的表达,细胞死亡率降低;同时抑制piR-16744和Caspase-1的表达,焦亡相关蛋白的表达水平相较于单独抑制piR-16744组降低。研究表明piR-16744可通过依赖Caspase-1的经典焦亡信号通路调控NMCs焦亡。In order to investigate the effect and mechanism of piR-16744 in H/R-induced pyroptosis of NMCs,a H/R model of NMCs was constructed in vitro with the time gradient of hypoxia for 6 h,12 h and 24 h,and reoxygenation for 6 h.The expression of piR-16744 was detected by RT-qPCR.Then,the expression levels of pyroptosis-related proteins,such as GSDMD,GSDMD-C and Caspase-1 were explored through WB,as well as inflammatory factors IL-1β and IL-18.LDH and PI staining were used to detect cell mortality.The results show that NMCs undergone significant pyroptosis after 6/6 h of H/R.Knockdown of piR-16744 results in the upregulation of pyroptosis-related proteins expression and cell mortality increase.Overexpression of piR-16744 inhibits the expression level of pyroptosis-related proteins and decreases the cell mortality.Compared to knocking down piR-16744 alone,the expression level of pyrolysis-related proteins is reduced after knockdown of piR-16744 and Caspase-1.piR-16744 regulates NMCs pyroptosis through the Caspase-1-dependent classical pyroptosis signaling pathway.

关 键 词：PIRNA 细胞焦亡 小鼠原代心肌细胞 GSDMD 心肌损伤 

分 类 号：R542.2[医药卫生—心血管疾病]