FOXO4对脂多糖诱导的心肌细胞凋亡的抑制作用  

Inhibitory Effect of FOXO4 on Lipopolysaccharide-induced Apoptosis in Cardiomyocytes

作　　者：李鸿渐 王思月 LI Hongjian;WANG Siyue(Department of Cardiology,People’s Hospital Affiliated to Shandong First Medical University,Jinan,271199,China)

机构地区：[1]山东第一医科大学附属人民医院心内科,济南市271199

出　　处：《医学分子生物学杂志》2024年第6期508-514,共7页Journal of Medical Molecular Biology

基　　金：山东省中医药科技项目(No.2021M0244)。

摘　　要：目的探讨叉头盒O4(forkhead box O4,FOXO4)对脂多糖(lipopolysaccharide,LPS)诱导的心肌细胞炎症反应和细胞凋亡的影响,并初步探讨潜在的分子机制。方法将LPS处理的H9C2细胞分别用FOXO4重组慢病毒载体(LPS+Lv-FOXO4)、对照载体(LPS+Lv-NC组)和/或nigericin(LPS+Lv-FOXO4+N组)处理FOXO4。实时定量聚合酶链式反应(RT-qPCR)检测FOXO4 mRNA表达。酶联免疫吸附实验检测细胞上清液中TNF-α和IL-1β的水平。流式细胞术检测细胞凋亡。蛋白质印迹分析FOXO4和NF-κB/NLRP3通路相关蛋白表达。结果LPS处理能够抑制心肌细胞中FOXO4的表达(P<0.05),而FOXO4过表达能够降低LPS处理的心肌细胞中TNF-α和IL-1β水平,抑制心肌细胞凋亡,降低促凋亡蛋白Bax表达并增加抗凋亡蛋白Bcl-2表达(P<0.05)。此外,FOXO4过表达能够通过抑制P65、IκBα磷酸化和P65核移位来抑制NF-κB信号通路活化(P<0.05)。FOXO4过表达可通过抑制NLRP3、ASC、cleaved caspase-1和N-GSDMD蛋白表达来抑制NLRP3炎症小体活化和细胞焦亡(P<0.05),而NLRP3炎症小体激活剂尼日利亚菌素能够逆转FOXO4过表达对LPS诱导心肌细胞损伤的保护作用(P<0.05)。结论FOXO4过表达可保护心肌细胞免受LPS刺激引起的炎症和凋亡,这可能是通过抑制NF-κB/NLRP3炎症小体信号通路介导的细胞焦亡来实现。Objective This study was to investigate the effects of FOXO4 on lipopolysaccharide(LPS)-induced inflammation and apoptosis of cardiomyocytes,and to explore the potential molecular mechanism.Methods LPS treated H9C2 cells were treated with FOXO4 recombinant lentiviral vector(LPS+Lv-FOXO4),control vector(LPS+Lv-NC group)and/or nigericin(LPS+Lv-FOXO4+N group)respectively.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of FOXO4 mRNA.The levels of TNF-αand IL-1βin cell supernatant were detected by enzyme-linked immunosorbent assay.The apoptosis was detected by flow cytometry.The expression levels of FOXO4 and NF-κB/NLRP3 pathway-related proteins was analyzed by Western blotting.Results LPS treatment could inhibit the expression of FOXO4 in myocytes(P<0.05),while overexpression of FOXO4 could decrease the levels of TNF-αand IL-1β,inhibit the apoptosis of myocytes,decrease the expression level of pro-apoptotic protein Bax and increase the expression level of anti-apoptotic protein Bcl-2 in myocytes treated with LPS(P<0.05).In addition,overexpression of FOXO4 could inhibit the activation of NF-κB signaling pathway by inhibiting P65,IκBαphosphorylation and P65 nuclear shift(P<0.05).Overexpression of FOXO4 inhibited NLRP3 inflammasome activation and pyroptosis by inhibiting NLRP3,ASC,cleaved caspase-1,and N-GSDMD protein expression(P<0.05).The NLRP3 inflammasome activator Nigeritin could reverse the protective effect of FOXO4 overexpression on LPS-induced myocardial cell injury(P<0.05).Conclusion Overexpression of FOXO4 can protect cardiomyocytes from LPS-induced inflammation and apoptosis,possibly by inhibiting pyroptosis mediated by NF-κB/NLRP3 inflammasome signaling pathway.

关 键 词：FOXO4 心肌细胞 炎症 凋亡 焦亡 NLRP3炎症小体 NF-ΚB 

分 类 号：R543.3[医药卫生—心血管疾病]