CREG1蛋白对血管紧张素Ⅱ诱导的小鼠心功能损伤的影响  

Effect of cellular repressor of E1A stimulated genes(CREG1) on cardiac function injury induced by angiotensin Ⅱ in mice

作　　者：宋海旭[1] 李洋[2] 孙鸣宇[2] 彭程飞[2] 田孝祥[2] 闫承慧[2] 韩雅玲[2] 

机构地区：[1]第四军医大学西京医院心内科,西安710032 [2]沈阳军区总医院心内科,沈阳110016

出　　处：《解放军医学杂志》2015年第1期16-21,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金重点项目(81130072);面上项目(81370243);国家"十二五"重大新药创制研究项目(2012ZX09303016-002)~~

摘　　要：目的探讨E1A激活基因阻遏子(CREG1)蛋白能否改善心肌纤维化小鼠的心功能。方法应用基因打靶方法建立广泛性基因敲除的CREG1杂合子小鼠和CREG1野生型小鼠模型。应用血管紧张素Ⅱ(AngⅡ)皮下埋泵方法建立小鼠心肌纤维化损伤模型,给予AngⅡ刺激14d后,采用HE和Masson染色检测小鼠心肌纤维化情况。应用Western blotting和免疫组化染色技术检测给予AngⅡ前及3、7、14d后两组小鼠心肌中CREG1蛋白的表达,并于给予AngⅡ14d后应用小动物超声仪检测心功能情况。AngⅡ给药同时,以皮下埋泵方式分别给予15、30、60、300μg/(kg·d)的外源性重组CREG1蛋白(治疗组)和生理盐水(对照组)14d,检测心功能,并应用TUNEL染色和Western blotting检测心肌凋亡情况。结果 Western blotting和免疫组化检测结果显示,未给予AngⅡ刺激时杂合子小鼠心肌中CREG1蛋白表达明显低于野生型小鼠(P<0.05)。给予AngⅡ刺激3、7、14d时,野生型小鼠和杂合子小鼠心肌中CREG1蛋白表达均明显下降(P<0.05),但杂合子小鼠下降更为显著(P<0.01);HE和Masson染色显示杂合子小鼠心肌纤维化程度较野生型小鼠严重,二者心功能明显下降,且杂合子小鼠心功能下降更为明显(P<0.05)。给予外源性重组CREG1蛋白治疗后,治疗组心功能较对照组明显改善(P<0.05),心肌凋亡数量明显下降(P<0.05)。结论在AngⅡ引起的小鼠心肌纤维化模型中,CREG1蛋白减少可使小鼠心功能损伤加重,给予外源性重组CREG1蛋白可明显改善心功能。Objective To investigate the effect of cellular repressor of E1A stimulated genes （CREG 1） on cardiac function in mouse with myocardial fibrosis. Methods CREG1 knockout mice （CREG1＋/- ） and CREG1 wild-type mice （CREG1＋/＋） were used to reproduce the model of myocardial fibrosis by subcutaneous pump burying of angiotensin Ⅱ fang 11 ）. After being stimulated with Ang Ⅱ for 14 days, myocardial fibrosis was verified by HE staining and Masson trichrome staining. Western blotting and immunohistochemistry were used to detect the expression of CREG1 in myocardium before stimulation and 3, 7, 14 days after the Ang 11 stimulation. The cardiac function was evaluated by echocardiography after Ang Ⅱ stimulation for 14 days. The CREG＋/＋ mice were given Ang Ⅱ for 14 days, and at the same time recombinant CREG1 protein [respectively 15, 30, 60 and 300μg/（kg·d）, intraperitoneal （IP） injections] （treatment group） and NaC1 （control group） were administered for treatment, and then cardiac function and myocardiac apoptosis were examined. Results Western blotting and immunohistochemistry showed that the expression of CREG1 in heart tissue was significantly lower in CILEG＋/- mice than in CREG＋/＋ mice （P〈0.05）. After Ang Ⅱ stimulation for 3, 7 and 14 days, the expression of CREG1 in heart tissue declined significantly in both CREG＋/ and CREG＋/＋ mice （P〈0.05）, especially in CREG＋/- mice （P〈0.01）. With HE and Masson staining, it was also found that CREG1 deficiency aggravated myocardial fibrosis and cardiac function deterioration in response to Aug Ⅱ stimulation （P〈0.05）. Conversely, exogenous infusion of recombinant CREG1 protein significantly inhibited the occurrence of myocardial apoptosis （P〈0.05）, thus ameliorated cardiac function （P〈0.05）. Conclusions CREG1 deficiency may aggravate the deterioration of cardiac function in mouse with myocardial fibrosis induced by Aug Ⅱ stimulation. The deterioration of cardiac function can be imp

关 键 词：E1A激活基因阻遏子 心肌纤维化 心功能 

分 类 号：R542.23[医药卫生—心血管疾病]