摘要
目的 探讨维立西呱对脂多糖(LPS)诱导的大鼠心肌细胞炎症反应和细胞凋亡的影响以及其对环状RNA PRKCI(circ PRKCI)表达的调控机制。方法 取大鼠心肌细胞H9C2,采用LPS诱导方式构建心肌细胞损伤模型;按随机数字表法分为对照组、LPS组、LPS+维立西呱-L组(1μmol/L)、LPS+维立西呱-M组(3μmol/L)、LPS+维立西呱-H组(10μmol/L)、LPS+空载体质粒(p c DNA)组、LPS+c irc PRKCI过表达载体(p c DNA-c irc PRKCI)组、LPS+维立西呱-H+阴性对照(s h-NC)组、LPS+维立西呱-H+circ PRKCI慢病毒短发夹RNA(sh-circ PRKCI)组。检测并比较各组H9C2细胞肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)等炎症因子水平,细胞凋亡率和裂解的胱天蛋白酶(Cleaved Caspase)-3、Cleaved Ca s p a s e-9等凋亡相关蛋白表达水平,以及c irc PRKCI mRNA表达水平。结果 与对照组比较,LPS组TNF-α水平、IL-6水平、细胞凋亡率及凋亡相关蛋白Cle a ve d Ca s p a s e-3、Cle a ve d Ca s p a s e-9蛋白表达水平均明显升高(均P<0.05);与LPS组比较,LPS+维立西呱-L组、LPS+维立西呱-M组、LPS+维立西呱-H组上述指标均明显降低(均P<0.05),且LPS+维立西呱-L组>LPS+维立西呱-M组>LPS+维立西呱-H组(均P<0.05)。与对照组比较,LPS组c irc PRKCI mRNA表达水平明显降低(P<0.05);与LPS组比较,LPS+维立西呱-L组、LPS+维立西呱-M组、LPS+维立西呱-H组c irc PRKCI mRNA表达水平均明显升高(均P<0.05),且LPS+维立西呱-L组<LPS+维立西呱-M组<LPS+维立西呱-H组(均P<0.05)。与LPS+pc DNA组比较,LPS+pc DNA-circ PRKCI组circ PRKCI mRNA表达水平明显升高(P<0.05),而TNF-α水平、IL-6水平、细胞凋亡率以及凋亡相关蛋白Cle a ve d Ca s p a s e-3、Cle a ve d Ca s p a s e-9蛋白表达水平均明显降低(均P<0.05)。与LPS+维立西呱-H+sh-NC组比较,LPS+维立西呱-H+sh-circ PRKCI组circ PRKCI mRNA表达水平明显降低(P<0.05),而TNF-α水平、IL-6水平、细胞凋亡率以及凋亡相关蛋白Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平均明显升高(均P<0.05)。结论 维立西呱可抑制LPS诱导的大鼠心肌细胞炎症反应和细胞凋亡,其作用机制是上调circ PRKCI mRNA表达。
Corresponding author:HUA Congjun,E-mail:29703295@qq.com[Abstract]Objective To investigate the effects of vericiguat on lipopolysaccharide(LPS)-induced inflammatory response and apoptosis in rat cardiomyocytes H9C2 and to analyze its regulation of circular RNA PRKCI(circPRKCI)expression.Methods LPS stimulation of H9C2 cells was used to establish a myocardial cell injury model,and random groups were established:control group,LPS group,LPS+vericiguat-L group,LPS+vericiguat-M group,LPS+vericiguat-H group,LPS+pcDNA group,LPS+pcDNA-circPRKCI group,LPS+vericiguat-H+sh-NC group,LPS+vericiguat-H+sh-circPR KCI group.vericiguat-H+sh-NC group,LPS+vericiguat-H+sh-circP RKCI group.qRT-PCR pcDNAwas performed to detect the expression of circPRKCI.The expression levels of tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6)were detected by ELISA.Apoptosis rate was detected by flow cytometry.Results Compared to the control group,the expression levels of TNF-αand IL-6,apoptosis rate and the protein expression of cleavedcaspase3,cleaved-caspase9 were increased and the expression of circPRKCI was decreased in the LPS group(all P<0.05).Compared to the LPS group,the expression levels of TNF-αand IL-6,apoptosis rate were decreased,and the expression of circPRKCI was increased in the LPS+vericiguat-L,LPS+vericiguat-M and LPS+vericiguat-H groups,and they were dose-dependent(all P<0.05).The expression levels of TNF-αand IL-6,apoptosis rate were reduced in the LPS+pcDNA-circPRKCI group compared to the LPS+pcDNA group(all P<0.05).Compared to the LPS+vericiguat-H+sh-NC group,the expression levels of TNF-αand IL-6,apoptosis rate were increased in the LPS+vericiguat-H+shcircPRKCI group(all P<0.05).Conclusion Vericiguat could inhibit the inflammatory response,apoptosis and thus attenuate LPS-induced H9C2 cell injury in cardiomyocytes through upregulation of circPRKCI mRNA expression.
作者
华崇俊
华秀红
黄春艳
潘轶斌
HUA Congjun;HUA Xiuhong;HUANG Chunyan;PAN Yibin(Department of Cardiology,Jinhua Hospital,Zhejiang University School of Medicine(Jinhua Municipal Central Hospital),Jinhua 321000,China;不详)
出处
《心电与循环》
2023年第5期427-433,共7页
Journal of Electrocardiology and Circulation
