miR-129-5p在脓毒症诱导的心脏炎症和功能障碍中的作用  

Role of miR-129-5p in sepsis-induced cardiac inflammation and dysfunction

作　　者：华云峰 杨珺 余娜 HUA Yunfeng;YANG Jun;YU Na(School of Clinical Medicine,Jiangsu College of Health Professions,Nanjing 210036;Department of Gastroenterology,Liyang Branch,Jiangsu Provincial People's Hospital,Liyang 213376;Department of Ultrasound Medicine,Wuxi People's Hospital,Nanjing Medical University,Wuxi 214023,China)

机构地区：[1]江苏卫生健康职业学院临床医学院,南京210036 [2]江苏省人民医院溧阳分院消化内科,213376 [3]南京医科大学附属无锡人民医院超声医学科,214023

出　　处：《国际心血管病杂志》2024年第6期379-384,共6页International Journal of Cardiovascular Disease

基　　金：江苏省高等学校基础科学研究(21KJB400152)。

摘　　要：目的:探讨miR-129-5p在脓毒症诱导的心脏炎症和功能障碍中的作用。方法:24只8周龄的C57/BL6雄性小鼠按随机数字表法随机分为正常(Normal)组、脂多糖(LPS)组、LPS+空病毒对照(minic-NC)组、LPS+miR-129-5p慢病毒(miR-129-5p minic)组,每组6只,采用腹腔注射LPS建立脓毒症小鼠模型。采用免疫组织化学染色检测配对盒基因6(PAX6),全自动生化分析仪检测血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB),TUNEL分析检测心肌凋亡情况。体外采用LPS诱导原代心肌细胞,模拟脓毒症引起的心功能不全。将细胞分为对照(Control)组、LPS+inhibitor-NC+si-NC组、LPS+inhibitor-NC+si-PAX6组、LPS+si-NC+miR-129-5p inhibitor组、LPS+miR-129-5p inhibitor+si-PAX6组。采用TUNEL分析检测心肌细胞凋亡情况,Western blot检测B淋巴细胞瘤-2(Bcl-2)、PAX6和人锌指E盒结合同源盒2(ZEB2)蛋白表达,实时荧光定量PCR检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6和IL-1β水平。结果:动物实验结果表明,与Normal组相比,LPS组小鼠血清CK、CK-MB水平,心肌细胞横截面积,TUNEL阳性细胞、PAX6阳性细胞比例和PAX6 mRNA表达水平显著升高(P<0.05)。与LPS组相比,LPS+miR-129-5p minic组CK、CK-MB水平,心肌细胞横截面积,TUNEL阳性细胞、PAX6阳性细胞比例和PAX6 mRNA表达水平显著降低(P<0.05)。体外细胞实验结果表明,与Control组相比,LPS+inhibitor-NC+si-NC组TUNEL阳性细胞比例,PAX6、ZEB2蛋白表达水平,TNF-α、IL-6、IL-1βmRNA表达水平显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05)。与LPS+si-NC+miR-129-5p inhibitor组相比,LPS+miR-129-5p inhibitor+si-PAX6组TUNEL阳性细胞比例,PAX6、ZEB2蛋白表达水平,TNF-α、IL-6、IL-1βmRNA表达水平显著降低(P<0.05),Bcl-2蛋白表达水平显著升高(P<0.05)。结论:miR-129-5p可能通过调控PAX6/ZEB2信号通路控制心肌炎症反应,并参与调节脓毒症诱导的心脏功能障碍。Objective:This study aimed to assess the role of miR-129-5p expression in sepsis-induced cardiac inflammation and dysfunction.Methods:Twenty-four 8-week-old C57/BL6 male mice were randomly divided into 4 groups,including normal group,lipopolysaccharide(LPS)group,LPS+mini NC group,and LPS+miR-129-5p mini group(n=6 in each group).A sepsis mouse model was constructed by intraperitoneal injection of LPS.Myocardial injury was assessed by HE staining,immunohistochemistry for PAX6,as well as automatic biochemical analysis for serum levels of creatine kinase(CK)and CK-MB.Primary cardiomyocytes exposed to LPS were established to simulate sepsis-induced cardiac dysfunction in vitro,and were divided into control group,LPS+inhibitor-NC+si-NC group,LPS+inhibitor-NC+si-PAX6 group,LPS+si-NC+miR-129-5p inhibitor group,and LPS+miR-129-5p inhibitor+si-PAX6 group.The degree of apoptosis was analyzed by TUNEL,and the expression of Bcl-2,PAX6 and ZEB2 protein was assessed by Western blotting.Levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6 and IL-1βwere determined by real-time fluorescence quantitative PCR(RT-qPCR).Results:Compared with normal group,serum levels of CK and CK-MB,cross-sectional area of myocardial cells,percentage of TUNEL or PAX6 positive cells,and PAX6 mRNA expression were significantly higher in LPS group(P<0.05).Compared with LPS group,these measuremnts were lower in LPS+miR-129-5p minic group(P<0.05).Furthermore,compared with control group,the percentage of TUNEL positive cells,and expression of PAX6 and ZEB2 proteins and TNF-α,IL-6 and IL-1βmRNA were increased(P<0.05),whereas and the expression of Bcl-2 protein was decreased significantly in LPS+inhibitor-NC+si-NC group(P<0.05).Compared with LPS+si-NC+miR-129-5p inhibitor group,the percentage of TUNEL positve cells,and the expression levels of PAX6 and ZEB2 protein and TNF-α,IL-6 and IL-1βmRNA were reduced(P<0.05),and the expression of Bcl-2 protein was elevated significantly in LPS+miR-129-5p inhibitor+si-PAX6 group(all P<0.05).Conclusion:MiR-129-5

关 键 词：miR-129-5p PAX6/ZEB2信号通路 脓毒症 

分 类 号：R459.7[医药卫生—急诊医学]