α-1抗胰蛋白酶对缺氧/复氧小鼠心肌细胞凋亡的影响  

Effects ofα-1 antitrypsin on cardiomyocyte apoptosis in hypoxia/reoxygenation mice

作　　者：黄达[1] 刘燕[1] 黄兰松 岑团 刘文静[1] 潘兴寿[1] 黄照河[1] HUANG Da;LIU Yan;HUANG Lansong;CEN Tuan;LIU Wenjing;PAN Xingshou;HUANG Zhaohe(Department of Cardiovascular,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院附属医院心内科,广西百色533000

出　　处：《右江医学》2021年第8期572-577,共6页Chinese Youjiang Medical Journal

基　　金：广西卫生和计划生育委员会自筹经费项目(Z2016420,Z20180208);广西高校中青年教师基础能力提升项目(KY2016YB343)。

摘　　要：目的建立小鼠心肌细胞缺氧/复氧模型,观察α-1抗胰蛋白酶(α-1 antitrypsin,A1AT)对小鼠心肌细胞凋亡的影响。方法体外采用连二亚硫酸钠(Na_(2)S_(2)O_(4))4 mmol/L诱导的缺氧培养基培养小鼠心肌细胞,模拟缺氧/复氧模型,并筛选A1AT最佳干预浓度,筛选A1AT最佳干预浓度实验分为7个组:Blank组、Control组、H/R组、A1AT 10μmol/L组、A1AT 20μmol/L组、A1AT 40μmol/L组以及A1AT 80μmol/L组。筛选最佳干预时间实验分为8个组:Blank组、Control组、H/R组、A1AT 1 h组、A1AT 2 h组、A1AT 4 h组、A1AT 6 h组及A1AT 12 h组。之后A1AT干预实验分组:正常组(Control组)、缺氧/复氧组(H/R组)、α-1抗胰蛋白酶干预组(A1AT组)。利用CCK-8检测细胞功能、流式细胞仪检测细胞凋亡、免疫荧光及Western Blot检测caspase-3的表达。并用SPSS 22.0进行数据统计。结果A1AT最佳干预浓度为20μmol/L,最佳干预时间为4小时。流式细胞仪检测发现H/R组细胞凋亡率增加,A1AT组细胞凋亡率明显下降,Control组、H/R组及A1AT组三组比较差异有统计学意义(P<0.05)。免疫荧光检测发现H/R组caspase-3分子表达强阳性,与Control组及A1AT组比较,差异有统计学意义(P<0.05)。Western Blot检测发现H/R组caspase-3蛋白表达上调,与Control组及A1AT组比较,差异有统计学意义(P<0.05)。结论α-1抗胰蛋白酶对小鼠缺氧/复氧心肌细胞凋亡具有改善作用。Objective To establish a hypoxia/reoxygenation model of cardiomyocytes in mice,and observe the effect ofα-1 antitrypsin(A1AT)on cardiomyocyte apoptosis in mice.Methods Mouse cardiomyocytes were cultured in vitro by hypoxia medium induced by(Na 2S _(2)O_(4))4 mmol/L of sodium dithionite,hypoxia/reoxygenation model was simulated,and the best intervention concentration of A1AT was selected.The experiment of screening the best intervention concentration of A1AT was divided into 7 groups:blank group,control group,H/R group and A1AT 10μmol/L group,A1AT 20μmol/L group,A1AT 40μmol/L group and A1AT 80μmol/L group.The experiment of screening the best intervention time was divided into 8 groups:blank group,control group,H/R group,A1AT 1 h group,A1AT 2 h group,A1AT 4 h group,A1AT 6 h group and A1AT 12 h group.After that,A1AT intervention experiment was divided into normal group(control group),hypoxia reoxygenation group(H/R group)andα-1 antitrypsin intervention group(A1AT group).CCK-8 was used to detect cell function,flow cytometry was used to detect the expression of apoptosis,and immunofluorescence and Western Blot were used to detect the expression of caspase-3.In addition,SPSS 22.0 was used for data statistics.Results The best intervention concentration for A1AT was 20μmol/L,and the best intervention time was 4 hours.The results of flow cytometry showed that the apoptosis rate increased in the H/R group and decreased significantly in the A1AT group,and there was statistically significant difference among the control group,the H/R group and the A1AT group(P<0.05).The Immunofluorescence detection revealed that the expression of caspase-3 molecules in the H/R group was strong positive,and compared with the control group and the A1AT group,the difference was statistically significant(P<0.05).Western Blot detection revealed that the expression of caspase-3 molecules in the H/R group was up-regulated,and compared with the control group and the A1AT group,the difference was statistically significant(P<0.05).Conclusionα-

关 键 词：α-1抗胰蛋白酶 Na 2S 2O 4 缺氧/复氧 凋亡 CASPASE-3 

分 类 号：R363[医药卫生—病理学]