重组鼠MANF通过激活SIRT1/AMPK信号通路参与脓毒症诱导心肌损伤的保护作用  

Recombinant mouse MANF participates in the protection of myocardial injury induced by sepsis by activating SIRT1/AMPK signaling pathway

作　　者：何浩[1] 李成[1] 胡赛[1] 夏风强 张弛 王静[1] He Hao;Li Cheng;Hu Sai;Xia Fengqiang;Zhang Chi;Wang Jing(Dept of Critical Care Medicine,Changsha Fourth Hospital(Changsha Hospital Affiliated to Hunan Normal University),Changsha410006)

机构地区：[1]长沙市第四医院(湖南师范大学附属长沙医院)重症医学科,长沙410006

出　　处：《安徽医科大学学报》2024年第11期1981-1988,共8页Acta Universitatis Medicinalis Anhui

基　　金：湖南省自然科学基金项目(编号:2021JJ70156)。

摘　　要：目的探讨中脑星形胶质细胞来源的神经营养因子(MANF)通过激活SIRT1/AMPK信号通路在脓毒症诱导心肌损伤中的保护作用。方法48只小鼠随机分为4组:对照组、重组鼠MANF(rmMANF)组、盲肠结扎穿刺(CLP)组和CLP+rmMANF组,每组12只。CLP后8 h检测存活率、脓毒症评分、肛温、血生化指标、心肌损伤病理指标及内质网应激(ERS)相关蛋白表达。将H9C2细胞分为对照(Con)组、脂多糖(LPS)组、LPS+rmMANF组、LPS+rmMANF+EX527组和LPS+rmMANF+Cpd C组。LPS处理24 h,收集细胞,分析细胞中ERS蛋白表达和凋亡情况。结果与CLP组比较,CLP+rmMANF组脓毒症评分及血清乳酸脱氢酶(LDH)、肌酸激酶(CK)、天冬氨酸氨基转移酶(AST)、血尿素氮(BUN)水平降低(P<0.01),肛门温度、血清白蛋白(ALB)水平升高(P<0.05);CLP+rmMANF组心脏组织中MANF表达显著增加(P<0.01),葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)表达及TUNEL阳性细胞百分比显著降低(P<0.05)。体外实验中,LPS刺激下调了H9C2细胞中SIRT1和AMPK的表达,而rmMANF进一步增加了SIRT1和AMPK的表达水平。与LPS+rmMANF组比较,LPS+rmMANF+EX527组和LPS+rmMANF+Cpd C组H9C2细胞中GRP78、CHOP蛋白表达和细胞凋亡率增加(P<0.05)。结论rmMANF通过激活SIRT1/AMPK信号通路抑制脓毒症诱导心肌损伤相关ERS,进而保护心肌损伤。Objective To investigate the protective effect of neurotrophic factor(MANF)derived from midbrain astrocytes on myocardial injury induced by sepsis by activating SIRT1/AMPK signaling pathway.Methods 48 mice were randomly divided into 4 groups:control group,recombinant mouse MANF(rmMANF)group,cecal ligation and puncture(CLP)group and CLP+rmMANF group,with 12 mice in each group.The survival rate,sepsis score,anal temperature,blood biochemical indexes,pathological indexes of myocardial injury and the expression of endoplasmic reticulum stress(ERS)related proteins were detected 8 h after CLP.H9C2 cells were divided into control group(Con),LPS group,LPS+rmMANF group,LPS+rmMANF+EX527 group and LPS+rmMANF+Cpd C group.The cells were collected after 24 h treatment with LPS,and the expression of ERS protein and apoptosis in cells were analyzed.Results Compared with CLP group,the sepsis score and serum Lactate dehydrogenase(LDH),creatine kinase(CK),aspartateaminotransferase(AST)and blood urea nitrogen(BUN)levels in CLP+rmMANF group decreased significantly(P<0.01),and the anal temperature and serum albumin(ALB)levels increased significantly(P<0.05).Compared with CLP group,the expression of MANF in CLP+rmMANF group increased significantly(P<0.01),and the expression of glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP)and the percentage of TUNEL positive cells decreased significantly(P<0.05).In vitro,LPS stimulation down-regulated the expression of SIRT1 and AMPK in H9C2 cells,while rmMANF further increased the expression level of SIRT1 and AMPK.Compared with LPS+rmMANF group,the expression of GRP78 and CHOP protein and the apoptosis rate of H9C2 cells in LPS+rmMANF+EX527 group and LPS+rmMANF+Cpd C group increased significantly(P<0.05).Conclusion rmMANF inhibits ERS related to sepsis-induced myocardial injury by activating SIRT1/AMPK signaling pathway,thereby protecting myocardial injury.

关 键 词：内质网应激 中脑星形胶质细胞来源的神经营养因子 脓毒症 心肌损伤 

分 类 号：R459.7[医药卫生—急诊医学]