内质网过度应激在肺缺血/再灌注小鼠心肌损伤中的作用  

Effects of excessive endoplasmic reticulum stress on lung ischemia/reperfusion induced myocardial injury in mice

作　　者：项冰倩 高慧[1] 郝卯林[1] 戴雍月[1] 王万铁[1] XIANG Bing-qian;GAO Hui;HAO Mao-lin;DAI Yong-yue;WANG Wan-tie(Ischemia/Reperfusion Injury Research Institute, Wenzhou Medical University, Wenzhou 325035, Chin)

机构地区：[1]温州医科大学缺血/再灌注损伤研究所,浙江温州325035

出　　处：《中国应用生理学杂志》2018年第1期8-13,共6页Chinese Journal of Applied Physiology

基　　金：浙江省公益技术应用研究项目(2013C33168);浙江省医药卫生科技计划项目(2010KYA132);浙江省新苗人才计划项目(2014R413043);温州市公益性科技计划项目(Y20140652)

摘　　要：目的:探讨内质网过度应激与肺缺血/再灌注小鼠心肌损伤的关系。方法:雄性健康SPF级C57BL/6J小鼠40只,随机将其分为4组(n=10):假手术组(Sham组)、肺缺血/再灌注组(I/R组)、ERS通路激动剂衣霉素(TM)组、ERS通路抑制剂4-苯基丁酸(4-PBA)组。采用夹闭左侧肺门30 min再灌注180 min的方法制备肺缺血/再灌注损伤模型。Sham组仅行开胸处理,不夹闭肺门,机械通气210 min;TM组、4-PBA组分别于造模前30 min腹腔注射衣霉素1 mg/kg和4-苯基丁酸400 mg/kg。于再灌注180 min时眼眶取血行心肌酶检测,处死后取心肌组织,行光镜、TUNEL Caspase 3酶活性、RT-PCR和Western blot检测。结果:与Sham组比较,光镜下I/R组、TM组和4-PBA组心肌细胞均有损伤性变化,肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)活性及心肌细胞凋亡指数、天冬氨酸特异性半胱氨酸蛋白酶3(Caspase 3)酶活性升高,p-Jun氨基末端激酶(p-JNK)、Caspase 12、CCAAT增强子结合蛋白同源蛋白(CHOP)、葡萄糖调节蛋白78(GRP78)及其mRNA表达上调(P<0.01);与I/R组比较,TM组光镜下心肌细胞损伤加重,血清CK-MB、LDH活性及心肌细胞凋亡指数、Caspase 3酶活性升高,p-JNK、Caspase 12、CHOP和GRP78蛋白及mRNA表达增加(P<0.01),4-PBA组以上指标均下降,光镜下心肌细胞损伤减轻(P<0.01);与TM组比较,4-PBA组光镜下心肌细胞损伤减轻,血清CK-MB、LDH活性及心肌细胞凋亡指数、Caspase 3酶活性降低,p-JNK、Caspase 12、CHOP和GRP78蛋白及mRNA表达下降(P<0.01)。结论:内质网过度应激参与肺I/R诱发的心肌损伤,抑制内质网过度应激能减轻心脏损伤。Objective： To investigate the effects of excessive endoplasmic reticulum stress on lung ischemia/reperfusion（ I/R） induced myocardial injury in mice. Methods： Forty healthy SPF male C57 BL/6 J mice were divided into 4 groups randomly（ n = 10） ： sham operation group（ Sham group）,lung I/R group（ I/R group）,endoplasmic reticulum stress（ ERS） pathway agonist Tunicamycin group（ TM） and ERS inhibitor4-phenyl butyric acid group（ 4-PBA）. The model of lung I/R injury was established by clamping the left hilum of lung for 30 min followed by 180 min of reperfusion. In sham group,only sternotomy was performed,the hilum of lung was not clamped,and the mice were mechanically ventilated for 210 min. In TM and 4-PBA groups,TM 1 mg/kg and 4-PBA 400 mg/kg were injected intraperitoneally,respectively,at 30 min before establishment of the model. At 180 min of reperfusion,blood samples were collected from the orbit for determination of myocardial enzyme. The animals were then sacrificed,and hearts were removed for determination of light microscope,TUNEL,Caspase 3 enzymatic activity,real-time polymerase chain reaction and Western blot. Results： Compared with sham group,the cardiomyocytes had obvious damage under light microscope,and the serum creatine kinase-MB（ CK-MB） and lactic dehydrogenase（ LDH） activities,apoptosis index and Caspase 3 enzymatic activity were increased significantly,the expressions of p-Jun N-terminalkinase（ p-JNK）,Caspase 12,CCAAT/enhancer-binding protein homologous protein（ CHOP）and glucose regulated protein 78（ GRP78） protein and mRNA were up-regulated in I/R,TM and 4-PBA groups（ P〈0.01）. Compared with I/R group,the cardiomyocytes damage was obvious under light microscope,and the serum CK-MB and LDH activities,apoptosis index and Caspase 3 enzymatic activity were increased significantly,the expressions of p-JNK,Caspase 12,CHOP and GRP78 protein and mRNA were up-regulated in group TM; while all above changes were relieved in group 4-PBA（ P〈0.01）

关 键 词：肺 再灌注损伤 小鼠 心肌 内质网 应激 

分 类 号：R363[医药卫生—病理学]