心肌缺血再灌注损伤小鼠E1A激活基因阻遏子基因表达及其对心肌的保护作用  

Expression of cellular repressor of E1A-stimulated gene and its cardioprotection role in mice with myocardial ischemia/reperfusion injury

作　　者：贺文帅 赵兴胜 吴云 温晓俐 南景龙 HE Wen-shuai;ZHAO Xing-sheng;WU Yun;WEN Xiao-li;NAN Jing long(Department of Cardiology, People＇s Hospital of Inner Mongolia Autonomous Region, Huhhot 010017, China)

机构地区：[1]内蒙古自治区人民医院心血管内科,呼和浩特010017

出　　处：《中华实用诊断与治疗杂志》2018年第6期525-528,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：内蒙古自治区科技厅项目(20161108)

摘　　要：目的探讨心肌缺血再灌注损伤(myocardial ischemia/reperfusion injury,MIR)小鼠E1A激活基因阻遏子基因(cellular repressor of E1A-stimulated gene,CREG)表达变化,及其对损伤心肌的保护作用。方法 CREG基因半敲除C57BL/6模型小鼠18只(A组),36只未敲除CREG基因C57BL/6小鼠随机分为B、C组各18只,3组采用线栓法制备MIR模型。建模前B组皮下微渗透泵给予外源性重组CREG蛋白1.5mg/(kg·d),连续14d,A、C组给予等量生理盐水。MIR后2h各组均处死6只小鼠,采用Western blot法检测心肌CREG蛋白表达,反转录PCR法检测心肌CREG mRNA表达;MIR后12h各组均取6只小鼠检测左室射血分数(left ventricular ejection fraction,LVEF)、左心室舒张末期容积(left ventricular end-diastolic volume,LVEDV)、左心室收缩末期容积(left ventricular end-systolic volume,LVESV),血清肌酸激酶(creatine kinase,CK)、肌酸激酶同工酶(creatine kinase isoenzyme,CK-MB)、谷草转氨酶(glutamic-oxaloacetic transaminase,GOP)、乳酸脱氢酶(lactate dehydrogenase,LDH);MIR 24h检测各组剩余6只小鼠心肌细胞凋亡指数。结果 A组心肌组织CREG蛋白、CREG mRNA表达(0.33±0.07、0.51±0.09)低于B组(0.85±0.19、0.97±0.10)、C组(0.67±0.15、0.99±0.05)(P<0.05);C组CREG蛋白表达低于B组(P<0.05),CREG mRNA表达与B组比较差异无统计学意义(P>0.05);A组LVEF[(34.2±3.1)%]低于B、C组[(58.7±5.1)%、(41.4±4.5)%](P<0.05),LVEDV[(121.8±14.0)μL]、LVESV[(77.2±9.0)μL]高于B组[(93.1±7.7)、(43.8±6.1)μL]、C组[(95.0±8.5)、(45.2±5.4)μL](P<0.05),C组LVEF低于B组(P<0.05);A组血清CK[(1 481.6±339.2)]u/L、CK-MB[(539.5±54.0)u/L]、GOP[(519.8±43.2)u/L]、LDH[(923.8±211.3)u/L]高于B组[(682.3±155.9)、(458.7±44.4)、(431.2±29.7)、(401.4±98.6)u/L]、C组[(874.0±193.5)、(489.1±47.1)、(460.0±32.5)、(600.5±115.8)u/L](P<0.05),且C组各指标均高于B组(P<0.05);A组心肌细胞凋亡指数[(57.21±2.26)%]明显高于B、C组[(16.94±1.75)%、(26.16±1.90)%](P<0.05),C组高于B组(PObjective To investigate the expression of cellular repressor of E1 A-stimulated gene（CREG）in mice with myocardial ischemia/reperfusion injury（MIR）and its protective effect on injured myocardium.Methods The MIR models were established by suture method in 18 C57 BL/6 mice with CREG gene semi-knock-out（group A）and 36 C57 BL/6 mice without CREG gene knock-out which were randomly divided into group B（n=18）and C（n=18）.Group B was administrated with exogenous recombinant CREG protein 1.5 mg/（kg·d）before the establishment of models by subcutaneous microosmotic pump,totally for 14 d.Group A and C were given the equivalent volume of normal saline.Six mice from each group were sacrificed 2 hafter MIR.The expression of GREG protein and mRNA were detected by Western blot and reverse transcription-PCR method.The levels of left ventricular ejection fraction（LVEF）,left ventricular end-diastolic volume（LVEDV）,left ventricular end-systolic volume（LVESV）,creatine kinase（CK）,creatine kinase isoenzyme（CK-MB）,glutamic-oxaloacetic transaminase（GOP）and lactate dehydrogenase（LDH）were detected 12 hafter MIR in 6 mice from each group.The myocardial apoptosis index was measured in the remained 6 mice from each group 24 hafter MIR.Results The levels of CREG protein and mRNA were significantly lower in group A（0.33±0.07,0.51±0.09）than those in group B（0.85±0.19,0.97±0.10）and C（0.67±0.15,0.99±0.05）（P〈0.05）,the level of CREG was significantly lower in group C than that in group B（P〈0.05）,and there was no significant difference in CREG mRNA level between group C and B（P〉0.05）.LVEF was significantly lower in group A（（34.2±3.1）%）than that in group B（（58.7±5.1）%）and group C（（41.4±4.5）%）（P〈0.05）,LVEDV and LVESV were significantly higher in group A（（121.8±14）,（77.2±9.0）μL）than those in group B（（93.1±7.7）,（43.8±6.1）μL）and group C（（95.0±8.5）,（45.2±5.4）μL）（P〈0.05）,and LVEF was signi

关 键 词：心肌缺血再灌注 心肌损伤 E1A激活基因阻遏子基因 保护作用 小鼠 

分 类 号：R54[医药卫生—心血管疾病]