NOD样受体家族蛋白3/半胱氨酸天冬氨酸蛋白酶3在大鼠心肌缺血再灌注损伤中的作用  

Role of NLRP3 Caspase-3 in myocardial ischemia reperfusion injury in rats

作　　者：薛同能 姚毅仁 丁莹 谷阳[1] Xue Tongneng;Yao Yiren;Ding Ying;Gu Yang(Department of Cardiology,The Huaian Clinical College of Xuzhou Medical University,The Affiliated Huaian No.1 People’s Hospital of Nanjing Medical University,Husi’an 223300,Jiangsu Province,China)

机构地区：[1]徐州医科大学淮安临床学院、南京医科大学附属淮安第一医院心内科,223300

出　　处：《中华老年心脑血管病杂志》2023年第12期1354-1358,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：淮安市自然科学研究计划项目(HAB202025);徐州医科大学附属医院优秀人才基金(XYFY2021030)。

摘　　要：目的探究NOD样受体家族蛋白3(NLRP3)/半胱氨酸天冬氨酸蛋白酶3(Caspase-3)在大鼠心肌缺血再灌注损伤诱导的心肌细胞凋亡中的作用及其对自噬的影响。方法选择SPF级雄性大鼠60只,建立心肌缺血再灌注损伤模型,将大鼠分为假手术组、模型组及尼莫地平组,每组20只。检测各组大鼠缺血再灌注后心功能,测定心肌酶、细胞因子。采用苏木精-伊红染色检测心肌病理改变;流式细胞术测定细胞凋亡率;透射电镜下计数自噬小体,RT-PCR和Westernblot法检测NLRP3和Caspase-3表达。结果各组大鼠左心室舒张末期压力、左心室收缩压、左心室压力最大上升及下降速率、心肌细胞凋亡率、TNF-α、白细胞介素6、肌酸激酶、天冬氨酸转氨酶和乳酸脱氢酶水平比较,差异有统计学意义(P<0.01)。模型组和尼莫地平组自噬小体计数明显高于假手术组[(10.55±1.87)个、(6.32±1.43)个vs(3.45±0.67)个,P<0.01],且尼莫地平组自噬小体计数明显低于模型组,差异有统计学意义(P<0.01)。与假手术组比较,模型组和尼莫地平组大鼠NLRP3、Caspase-3mRNA及蛋白表达明显升高,差异有统计学意义(P<0.01);与模型组比较,尼莫地平组大鼠NLRP3、Caspase-3mRNA及蛋白表达明显降低,差异有统计学意义(P<0.01)。结论大鼠缺血再灌注损伤可增加细胞凋亡率,降低心功能,诱发炎性反应,增强机体自噬小体形成能力,与NLRP3/Caspase-3的异常高表达有关。Objective To explore the roee of NLRP3/Caspase-3 in myocardial apoptosis induced by ischemia/reperfusion (IR)injury and its eefect on myocardiocyte autophagy in rats.Methods A total of 60 SPF-grade maee rats were randomly divided into sham operation,model,and nimodip-ine treatment groups,with 20 rats in each group.Rat model of myocardial IR injury was estab-lished in the rats of the two latter groups.Cardiac function was assessed,and the levels of myocar-dial enzymes and cytokines were measured.Additionally,myocardial pathological changes were detected using HE staining.Furthermore,flow cytometry was utilized to determine the apoptotic rate of myocardiocytes,and the autophagosomes were counted under transmission electron micro-scope.Moreover,the expression of NLRP3 and Caspase-3 was measured using RT-PCR and Western blotting.Results Significant differences were observed in lett ventricular end diastolic pressure,lett ventricular systolic pressure,maximal rate of rise and fall in lett ventricular pressure,ap-optotic rate of myocardial cells,and levels of TNF-α,1L-6,CK,AST and LDH in the three groups(P<0.01).Notably,both the model group and nimodipine treatment group exhibited significantly higher autophagosome than the sham operation group(10.55±1.87 and 6.32±1.43 vs 3.45±0.67 units,P<0.01),and the nimodipine group displayed a significantly lower autophagosome count than the model group(P<0.01).The mRNA and protein levels of NLRP3 and Caspase-3 were notably higher in the model group and nimodipine group than the sham operation group(P<0.01),and in the model group than the nimodipine group(P<0.01).Conclusion Myocardial 1R injury in rats can increase myocardiocyte apoptosis,reduce cardiac function,induce inflammatory response,and enhance autophagosome formation,which is related to the abnormal high expression of NLRP3/Caspase-3.

关 键 词：心肌再灌注损伤 NLR蛋白质类 半胱氨酸天冬氨酸蛋白酶 肌细胞 心脏 细胞凋亡 

分 类 号：R542.22[医药卫生—心血管疾病]