大鼠急性心肌缺血再灌注损伤中Cx43、ERα的表达及性别差异  

Expression and Sex Difference of Cx43 and ERαin Rats with Acute Myocardial Ischemia-reperfusion Injury

作　　者：陈亚敏 赵明慧 薛凌 CHEN Yamin;ZHAO Minghui;XUE Ling(Third Department of Cardiovascular Medicine,Third Affiliated Hospital of Jinzhou Medical University,Jingzhou Liaoning 121000,China)

机构地区：[1]锦州医科大学附属第三医院心血管内三科,辽宁锦州121000

出　　处：《华南国防医学杂志》2022年第4期248-252,共5页Military Medical Journal of South China

基　　金：辽宁省自然科学基金(20170540372);锦州医科大学附属第三医院青苗院长基金(FSSYQM2020004)。

摘　　要：目的研究大鼠急性心肌缺血再灌注损伤(myocardial ischemia-reperfusioninjury,MI/RI)中缝隙连接蛋白43(connexins43,Cx43)、雌激素受体α(estrogenreceptorα,ERα)的表达及性别差异。方法32只SD大鼠依据随机数字表法分为4组:假手术雄性组、假手术雌性组、MI/RI雄性组、MI/RI雌性组,建立MI/RI模型,每组8只大鼠。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)测定大鼠血清肌酸激酶同工酶MB(creatine kinaseMB,CK-MB)、乳酸脱氢酶(lactatedehydrogenase,LDH)的浓度。采用苏木精-伊红(hematoxylin-eosin,HE)染色法观察大鼠心肌细胞的形态学变化;免疫组织化学法(immunohistochemistry,IHC)检测Cx43、ERα的表达;蛋白质免疫印迹法(Western blot)检测心肌细胞Cx43、B淋巴细胞瘤-2基因(B-cell lymphoma-2,BCL-2)、BCL2-相关X蛋白(BCL2-associ-atedX,BAX)的表达。结果MI/RI雄性组CK-MB与LDH的浓度均高于MI/RI雌性组,差异有统计学意义(P<0.05)。HE染色结果显示:MI/RI雄性组与MI/RI雌性组相比,心肌纤维断裂较多,且心肌纤维间充血、出血较严重。IHC结果显示:Cx43蛋白大部分位于缝隙连接处和心肌细胞膜,与假手术组比较,MI/RI雄性组与MI/RI雌性组Cx43阳性表达减少(P<0.001),MI/RI雄性组阳性表达明显少于MI/RI雌性组(P<0.001)。ERα主要位于心肌细胞核,假手术雌性组与假手术雄性组阳性表达无明显差别,MI/RI雄性组ERα阳性表达少于MI/RI雌性组(P<0.001)。West-ernblot结果显示:与MI/RI雄性组相比,MI/RI雌性组Cx43增高,BAX/BCL-2表达降低,组间比较差异有统计学意义(P均<0.001)。结论大鼠在急性MI/RI中Cx43的表达存在性别差异,在MI/RI雌性组表达较多,并且可能与ERα有关,另外,Cx43对急性MI/RI有保护作用。Objective To study the expression of connexins 43(Cx43)and estrogen receptorα(ERα)and gender differences in acute myocardial ischemia-reperfusion injury(MI/RI)in rats.Methods A total of 32 SD rats were divided into 4 groups according to the random number table methods;sham-operated male group,sham-operated female group,MI/RI male group,MI/RI female group and MI/RI model were established,with 8 rats in each group.The concentrations of serum creatine kinase MB(CK-MB)and lactate dehydrogenase(LDH)in rats were determined by enzyme-linked immunosorbent assay(ELISA).The morphological changes of rat cardiomyocytes were observed by hematoxylin-eosin(HE)staining;the expressions of Cx43 and ERαwere detected by immunohistochemistry(IHC);the expressions of Cx43,B-cell lymphoma-2(BCL-2)and BCL2-associated X(BAX)in cardiomyocytes were detected by Western blot.Results The concentrations of CK-MB and LDH in the MI/RI male group were higher than those in the MI/RI female group,and the difference was statistically significant(P<0.05).HE staining showed that compared with the MI/RI female group,the MI/RI male group had more myocardial fiber breakage,and the myocardial fiber congestion and bleeding were more serious.IHC showed that most of the Cx43 protein was located in the gap junction and myocardial cell membrane.Compared with the sham-operated group,the positive expression of Cx43 in the MI/RI male group and MI/RI female group was reduced(P<0.001),and the positive expression in the MI/RI male group was significantly less than that in the MI/RI female group(P<0.001).ERαwas mainly located in the nucleus of myocardial cells.There was no significant difference in the positive expression of females and males in the sham-operated group.The positive expression of ERαin the MI/RI male group was less than that in the MI/RI female group(P<0.001).Western blot showed that compared with the MI/RI male group,the expression of Cx43 of MI/RI female group increased but BAX/BCL-2 decreased,and the differences were statistically significant(a

关 键 词：心肌缺血再灌注损伤 缝隙连接蛋白43 雌激素受体α BCL2-相关X蛋白 B淋巴细胞瘤-2基因 

分 类 号：R541[医药卫生—心血管疾病]