Flt3基因体外转染对H9c2大鼠心肌细胞缺血再灌注损伤的影响  

Effect of Flt3 gene transfection on ischemia reperfusion injury in H9c2 rat cardiomyocytes

作　　者：蒋春英[1] 邹艳 魏鹏[1] 张苗苗[1] 蒋鹤 嵇利亚[1] JIANG Chunying;ZOU Yan;WEI Peng;ZHANG Miaomiao;JIANG He;JI Liya(Department of Cardiology,Xuzhou Central Hospital,Xuzhou Institute of Cardiovascular Disease,Xuzhou,Jiangsu 221009,China)

机构地区：[1]徐州市中心医院心内科,徐州市心血管病研究所,江苏徐州221009

出　　处：《徐州医科大学学报》2021年第10期737-741,共5页Journal of Xuzhou Medical University

基　　金：徐州市科学技术局应用基础研究计划项目(KC17101);江苏省博士后科研资助计划(2020Z44)。

摘　　要：目的探讨腺病毒转染Flt3基因对H9c2大鼠心肌细胞缺血再灌注损伤的影响。方法建立H9c2大鼠心肌细胞模拟缺血再灌注模型。Flt3基因腺病毒转染H9c2大鼠心肌细胞,缺血再灌注后免疫荧光法验证转染效果。将H9c2大鼠心肌细胞分为正常对照组(C组)、缺氧复氧组(IRI组)、空载体组(Ad-GFP组)及基因转染组(Ad-Flt3组)。Western blot法检测Flt3蛋白、Caspase-3蛋白的表达。采用CCK-8法检测各组心肌细胞活性。比色法测定乳酸脱氧酶(LDH)、肌酸激酶同工酶(CK-MB)、丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性。结果与C组比较,心肌细胞缺血再灌注损伤后Flt3蛋白表达下降(P<0.05),Caspase-3蛋白表达明显增加(P<0.01),细胞活力显著下降(P<0.05)。转染Flt3基因后心肌细胞中的Flt3蛋白较未转染细胞显著增加(P<0.05),Caspase-3蛋白表达显著下降(P<0.05),细胞活力显著升高(P<0.01)。与C组比较,缺血再灌注后细胞上清液LDH、CK-MB、MDA含量显著升高,SOD活性显著下降(P<0.05或P<0.01),而Flt3基因转染后细胞上清液LDH、CK-MB、MDA含量较未转染细胞显著降低,SOD活性升高(P<0.05或P<0.01)。结论Flt3基因转染H9c2大鼠心肌细胞可减轻细胞缺血再灌注损伤,其机制可能与抗氧化应激及减轻细胞凋亡有关。Objective To explore the effects of FMS-like tyrosine kinase 3(Flt3)gene transfection on ischemia reperfusion injury(IRI)in H9c2 rat cardiomyocytes in vitro.Methods A model of IRI was established using H9c2 rat cardiomyocytes.Flt3 gene carried by adenovirus was transfected into H9c2 rat cardiomyocytes.After the model was established,the efficiency of Flt3 gene transfection was detected by immunofluorescence.H9c2 rat cardiomyocytes were divided into four groups:a control group,an IRI group,an Ad-GFP group and an Ad-Flt3 group.The expression of Flt3 and Caspase-3 protein was detected by Western blot.The viability of cardiomyocytes was detected by Cell Counting Kit-8.The levels of lactate dehydrogenase(LDH),creatine kinase MB isozyme(CK-MB),maiondialdehyde(MDA)and superoxide diamutase(SOD)in the supernatant of cell culture were measured by the colorimetric method.Results After IRI,the expression of Flt3 protein decreased,the expression of Caspase-3 protein increased and the viability of cardiomyocytes was significantly up-regulated,compared with the control group(P<0.05 or P<0.01).In comparison with non-transfected groups,Flt3 protein expression was enhanced and Caspase-3 protein expression was weakened after transfection of Flt3 gene,compared with the control group(P<0.05 or P<0.01).The viability of cardiomyocytes was up-regulated in Flt3 transfected group(P<0.01).After IRI,the levels of LDH,CK-MB and MDA obviously increased,while the activity of SOD reduced in the supernatant of cell culture,compared with the comtrol group(P<0.05 or P<0.01).However,the contents of LDH,CK-MB and MDA were lower in the Flt3 transfected group than those in the non-transfected groups while the activity of SOD was higher(P<0.05 or P<0.01).Conclusions Transfection of Flt3 gene can protect cultured H9c2 rat cardiomyocytes against IRI by inhibiting oxidative stress and reducing apoptosis.

关 键 词：FLT3基因 缺血再灌注损伤 转染 

分 类 号：R363[医药卫生—病理学]