SHH/GLI1信号通路调控巨噬细胞焦亡参与急性心肌梗死的分子机制  

Molecular Mechanism of SHH/GLI1 Signaling Pathway Regulating Macrophage Pyroptosis in Acute Myocardial Infarction

作　　者：廉明[1] 公方娜 赵王磊[1] LIAN Ming;GONG Fangna;ZHAO Wanglei(The Affiliated Hospital of Inner Mongolia Medical University,Inner Mongolia Hohhot 010059,China)

机构地区：[1]内蒙古医科大学附属医院急诊科,内蒙古呼和浩特010059

出　　处：《河北医学》2024年第12期1982-1987,共6页Hebei Medicine

基　　金：内蒙古自治区卫生健康委医疗卫生科技计划项目,(编号:202102112)。

摘　　要：目的:探究SHH/GLI1信号通路调控巨噬细胞焦亡在急性心肌梗死(AMI)中的可能分子机制。方法:将50只大鼠随机分为:假手术组、AMI组、抑制SHH组、清除巨噬细胞组、清除巨噬细胞+抑制SHH组,每组10只,收集各组大鼠心肌组织与血清。培养巨噬细胞Raw264.7分为:对照组、缺氧组、抑制SHH组。Masson染色检测心肌组织纤维化水平及梗死面积。Western blotting检测SHH、GLI1、NLRP3、Caspase-1及GSDMD蛋白水平。RT-qPCR检测SHH、GLI1、F4/80mRNA水平。ELISA试剂盒检测血清CK-MB、LDH含量。结果:与假手术组相比,AMI组大鼠心肌纤维化水平增加,梗死面积增加,心肌组织中SHH、GLI1蛋白表达增加,血清CK-MB、LDH含量增加(P<0.05),心肌组织中焦亡相关蛋白NLRP3、Caspase-1及GSDMD蛋白表达增加(P<0.05),清除巨噬细胞组大鼠心肌组织中F8/40mRNA水平减少(P<0.05),提示巨噬细胞清除成功;与AMI组相比,抑制SHH组大鼠心肌纤维化水平减少,梗死面积减少,心肌组织中SHH、GLI1、NLRP3、Caspase-1及GSDMD蛋白表达减少(P<0.05),清除巨噬细胞组大鼠血清CK-MB、LDH含量减少(P<0.05),清除巨噬细胞+抑制SHH组大鼠血清CK-MB、LDH含量与清除巨噬细胞组大鼠变化保持一致(P<0.05)。与对照组相比,缺氧组巨噬细胞中SHH、GLI1mRNA水平与NLRP3、Caspase-1及GSDMD蛋白表达增加(P<0.05);与缺氧组相比,抑制SHH组巨噬细胞中SHH、GLI1mRNA水平与NLRP3、Caspase-1及GSDMD蛋白表达减少(P<0.05)。结论:下调SHH/GLI1信号通路可抑制巨噬细胞焦亡改善AMI。Objective:To explore the possible molecular mechanism of sonic hedgehog(SHH)/glioma-associated oncogene 1(GLI1)signaling pathway regulating macrophage pyroptosis in acute myocardial infarction(AMI).Methods:Fifty rats were randomly divided into sham operation group,AMI group,SHH inhibition group,macrophage clearance group,macrophage clearance+SHH inhibition group,with 10 rats in each group.Myocardial tissue and serum were collected.Cultured macrophages Raw264.7 were divided into control group,hypoxia group and SHH inhibition group.Masson staining was used to detect myocardial fibrosis level and infarct size.The protein levels of SHH,GLI1,NLRP3(NOD-,LRR-,and pyrin domain-containing protein 3),gasdermin-D(GSDMD)were detected by Western blotting.The mRNA levels of SHH,GLI1 and F4/80 were detected by real-time reverse transcriptase-polymerase chain reaction(RT-qPCR).Serum creatine kinase-myocardial band(CK-MB)and lactate dehydrogenase(LDH)were detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with sham operation group,myocardial fibrosis level,infarct size,SHH and GLI1 protein expression,serum CK-MB and LDH contents were significantly increased in AMI group(P<0.05).The expressions of NLRP3,Caspase-1 and GSDMD proteins in myocardial tissue were significantly increased(P<0.05),and the F8/40 mRNA levels in myocardial tissue of macrophage clearance group were significantly decreased(P<0.05),indicating successful macrophage clearance.Compared with AMI group,the myocardial fibrosis level,infarct size and protein expressions of SHH,GLI1,NLRP3,Caspase-1 and GSDMD were significantly decreased in the inhibited SHH group(P<0.05),and the serum CK-MB and LDH contents in the macrophage clearance group were significantly decreased(P<0.05).The serum CK-MB and LDH contents of rats in the macrophage clearance+SHH inhibition group were compared to those in the macrophage clearance group(P<0.05).Compared with the control group,the mRNA levels of SHH and GLI1and the protein expressions of NLRP3,Caspase-1 and GSDMD in m

关 键 词：急性心肌梗死 SHH/GLI1信号通路 巨噬细胞 焦亡 

分 类 号：R542.22[医药卫生—心血管疾病]