七氟醚后处理联合右美托咪定对心肌缺血/再灌注损伤小鼠的心肌保护作用机制  

Cardioprotective Effects of Sevoflurane Postconditioning Combined with Dexmedetomidine on Myocardial Ischemia/Reperfusion Injury in Mice

作　　者：金辉[1] 肖志博[1] 葛树胜 吴小精 尹顺花 李媛[1] JIN Hui;XIAO Zhibo;GE Shusheng;WU Xiaojing;YIN Shunhua;LI Yuan(The First Affiliated Hospital of Hainan Medical University,Haikou 570100,Hainan,China)

机构地区：[1]海南医学院第一附属医院,海口570100 [2]海南省妇女儿童医学中心

出　　处：《中西医结合心脑血管病杂志》2023年第21期3908-3915,共8页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：海南省自然科学基金青年基金项目(No.820QN397)。

摘　　要：目的:探索七氟醚(Sev)后处理(Sev-postC)对右美托咪定(Dex)心肌保护的协同增效作用及其分子机制。方法:将20只31周龄雄性C57BL/6J小鼠随机分为Sham组、缺血/再灌注损伤模型(Model)组、Dex组、Dex+Sev-postC组,每组5只,采用苏木精-伊红(HE)染色和Masson染色检测小鼠心肌损伤情况;酶联免疫吸附试验(ELISA)测定小鼠血清肌钙蛋白I(cTnI)、乳酸脱氢酶(LDH)、脑钠肽(BNP)、肌酸激酶同工酶(CK-MB)水平。体外实验中将H9c2心肌细胞分为Control组、缺氧/复氧损伤模型(H/R)组、Dex组、Dex+Sev+siRNA NT组和Dex+Sev+巨噬细胞迁移抑制因子(MIF)siRNA组,或Dex+Sev+pcDNA3.1组和Dex+Sev+pcDNA3.1-MIF组,转染MIF siRNA敲低MIF/转染pcDNA3.1-MIF过表达MIF。蛋白免疫印迹法(Western Blot)检测敲低或过表达MIF的效果,CCK-8法测定细胞存活能力;V-异硫氰酸荧光素(annexin V-FITC)/碘化丙啶(PI)双标记流式细胞术测定细胞凋亡率。结果:小鼠在体实验HE染色与Masson染色结果显示,与Sham组比较,Model组小鼠心肌纤维断裂溶解,梗死处有炎性细胞浸润;心肌纤维化程度明显,胶原蛋白增加;与Model组比较,Dex组和Dex+Sev-postC组心肌损伤明显改善。ELISA检测结果显示,与Sham组比较,Model组血清cTnI、LDH、BNP和CK-MB水平均明显上升(P<0.01);与Model组比较,Dex组、Dex+Sev-postC组血清cTnI、LDH、BNP和CK-MB水平均明显下调(P<0.05或P<0.01);与Dex组比较,Dex+Sev-postC组下调更明显(P<0.05或P<0.01)。体外细胞实验结果显示,Western Blot结果证明在H9c2细胞中成功敲低或过表达MIF。与Control组比较,H/R组细胞存活能力明显降低(P<0.01),细胞凋亡率明显升高(P<0.01)。与H/R组比较,Dex组和Dex+Sev+MIF siRNA组细胞存活能力明显上升(P<0.05),细胞凋亡率明显降低(P<0.01);Dex+Sev+siRNA NT组、Dex+Sev+pcDNA3.1组和Dex+Sev+pcDNA3.1-MIF组心肌细胞存活能力明显上升(P<0.05或P<0.01),细胞凋亡率明显降低(P<0.01),且与Dex+Sev+siRNA NT组比较,DObjective:To explore the protective effect of sevoflurane postconditioning combined with dexmedetomidine on myocardiaum protective function.Methods:Twenty 31-week-old male C57BL/6J mice(specific pathogen free grade)were randomly divided into 4 groups:sham group,ischemia/reperfusion injury(model)group,dexmedetomidine(Dex)group,sevoflurane postconditioning combined with dexmedetomidine(Dex+Sev-postC)group,with 5 mice in each group.Hematoxylin-eosin(HE)staining and Masson staining were used to detect myocardial injury in mice.Enzyme linked immunosorbent assay(ELISA)was used to determine the levels of cardiac troponin I(cTnI),lactic dehydrogenase(LDH),brain natriuretic peptide(BNP),and creatine kinase-MB(CK-MB)in mice serum.In vitro experiments were conducted by using H9c2 as cellular model.Cells were divided into five groups:control group,H/R group,Dex group,Dex+Sev+siRNA NT group,and Dex+Sev+MIF siRNA group or Dex+Sev+pcDNA3.1 group and Dex+Sev+pcDNA3.1-MIF group.Transfection with macrophage migration inhibitory factor(MIF)siRNA to knockdown the expression of MIF,and transfection of pcDNA3.1-MIF to over expression of MIF.Western Blot was used to detect the knockdown or overexpression efficiency.CCK-8 method was used to measure the cell viability.Annexin V-FITC/propidium iodide(PI)double-stained followed by flow cytometry was used to measure cell apoptosis rate.Results:In vivo HE staining and Masson staining results showed that compared with sham group,model group mice myocardial fibers broken and dissolved,inflammatory cells infiltrated in infarct area,the degree of myocardial fibrosis was obvious and the collagen deposition increased.Compared with model group,Dex group and Dex+Sev-postC group myocardial injury improved significantly.ELISA detection of serum cTnI,LDH,BNP,and CK-MB results showed that compared with sham group,the four indicators in model group significantly increased(P<0.01);compared with model group,the levels of 4 indicators in Dex group and Dex+Sev-postC group significantly down-regulated(P<0.05

关 键 词：缺血/再灌注损伤 右美托咪定 七氟醚 巨噬细胞迁移抑制因子 实验研究 

分 类 号：R614[医药卫生—麻醉学]