异氟烷预处理对缺氧/复氧引起的心肌细胞损伤的影响  

Effect of Isoflurane on anoxia/reoxygenation-induced cardiocyte injury

作　　者：付海钰 秦福恩[1] 徐帅 杨淋[1] 钟祖凌 巩固[1] FU Haiyu;QIN Fuen;XU Shuai;YANG Lin;ZHONG Zuling;GONG Gu(Department of Anesthesiology,General Hospital of Chengdu Military Region of PLA,Sichuan Province,Chengdu 610083,China)

机构地区：[1]成都军区总医院麻醉科,四川成都610083

出　　处：《中国医药导报》2018年第8期9-14,共6页China Medical Herald

基　　金：中央军委后勤保障部军队后勤科研计划项目(CCD16J001)

摘　　要：目的探讨异氟烷(Iso)在缺氧/复氧(A/R)诱导的心肌细胞损伤中的影响及其作用机制。方法培养心肌细胞H9c2,用不同浓度的Iso(1%、2%、3%)预处理后进行3 h的缺氧和3 h的复氧。MTT检测细胞生长,筛选Iso使用浓度,随后实时定量PCR检测miR-499表达。在3%Iso预处理的A/R损伤心肌细胞中转染miR-499的过表达试剂miR-499 mimic,流式细胞术检测细胞凋亡;Western blot检测Caspase-3、Caspase-9、Bax和Bcl-2的蛋白表达;试剂盒检测细胞中乳酸脱氢酶(LDH)、肌酸激酶(CK)和谷氨酸氨基转移酶(ALT)的含量。结果与Control组比较,A/R组的细胞活力显著下降(P<0.05),同时Iso预处理剂量依赖性的诱导细胞活力增加,由而筛选出3%作为Iso预处理浓度。与Control组比较,miR-499的表达在A/R组中显著提升(P<0.05),并在3%Iso预处理心肌细胞(Iso组)中下降(P<0.05)。与Control组比较,A/R组的细胞生长显著降低(P<0.05),细胞凋亡升高(P<0.05),促凋亡蛋白Caspase-3、Caspase-9、Bax表达升高(P<0.05),抑凋亡蛋白Bcl-2表达降低(P<0.05),LDH、CK和ALT的含量明显升高(P<0.05)。Iso预处理抑制以上由A/R诱导的H9c2细胞损伤,并且此心肌细胞保护作用可被miR-499过表达反转。结论 Iso可以通过抑制miR-499表达减轻A/R引起的心肌细胞损伤。Objective To investigate the effect of Isoflurane(Iso)on anoxia/reoxygenation(A/R)-induced cardiocyte injury and underlying mechanism.Methods Cultured cardiocytes(H9c2)were pre-treated with Iso(1%,2%,3%)and then stimulated with 3 h of anoxia and 3 h of reoxygenation.Cell proliferation was measured by MTT method,and the Iso concentration was selected.The miR-499 overexpression reagent,miR-499 mimic was transfected into cardiocytes pre-treated with 3%Iso and simulated with anoxia/reoxygenation.Cell apoptosis was detected by flow cytometry assay.The expression level of caspase-3,caspase-9,Bax and Bcl-2 were measured by Western blot.The contents of lactic dehydrogenase(LDH),creatine kinase(CK)and alanine aminotransferase(ALT)were detected by corresponding kit,respectively.Results Compared with the control group,cell proliferation was significantly decreased in A/R group(P<0.05),and increased dose-dependently in Iso pre-treated cells(P<0.05),thus 3%was selected to be the concentration of Iso pre-treatment.Compared with the control group,the expression of miR-499 was significantly increased in A/R group(P<0.05),and decreased in 3%Iso treated cells(P<0.05).Compared with the control group,cell proliferation was significantly decreased(P<0.05);cell apoptosis was increased(P<0.05);the expression of pro-apoptotic protein caspase-3,caspase-9 and Bax were increased(P<0.05),anti-apoptotic protein Bcl-2 was decreased(P<0.05);the content of LDH,CK and AST were increased in A/R group(P<0.05).Iso pre-treat inhibited A/R induced H9c2 injury,and the protective effect of Iso was reversed by miR-499 overexpression in damaged cardiocyte.Conclusion Isoflurane attenuates anoxia/reoxygenation-induced cardiocyte injury through inhibiting miR-499 expression.

关 键 词：异氟烷 miR-499 心肌细胞 缺氧/复氧 

分 类 号：R972[医药卫生—药品]