基质金属蛋白酶-9抑制剂治疗自身免疫性心肌炎大鼠模型的实验研究  

Effects of matrix metalloproteinase-9 inhibitor in Lewis rats with experimental autoimmune myocarditis

作　　者：韩丽娜[1] 李铁岭[2] 张亚晶[1] 杨庭树[1] 丁宇[1] 赵晓宁[1] 郭树理[1] 

机构地区：[1]中国人民解放军总医院南楼心血管一科,北京100853 [2]中国人民解放军总医院南楼干部理疗科,北京100853

出　　处：《中华心血管病杂志》2011年第2期118-123,共6页Chinese Journal of Cardiology

基　　金：2007年北京市优秀人才培养资助个人项目（20071D1600200391）;2008年北京市科技新星计划（B类Jf2008B54）.

摘　　要：目的 观察基质金属蛋白酶（MMP）-9抑制剂米诺环素对实验性自身免疫性心肌炎（EAM）Lewis大鼠模型的治疗效果，并探讨其机制。方法 Lewis大鼠60只，6～8周龄，体重150～170g。双足底注射心肌C蛋白片段和完全弗氏辅佐剂的油状混合物，腹腔注射百日咳毒素建立自身免疫性心肌炎大鼠模型。根据药物干预的时期不同将大鼠分为早期干预组、中期干预组和晚期干预组3组，每组20只，不同的干预组又分为米诺环素治疗组（治疗组）和对照组两个亚组，每个亚组10只。早期干预组治疗组为腹腔注射50me／kg的米诺环素，1次／d，连续21d，免疫注射后第1～2l天。中期干预组治疗组为腹腔注射50me／kg的米诺环素，1次／d，连续21d，免疫注射后第8～28天。晚期干预组治疗组为腹腔注射50mg／kg的米诺环素，1次／d，连续21d，免疫注射后第15～35天。各对照组均为在相同时间给予与治疗组相同体积的生理盐水腹腔注射。干预结束后，处死动物，心脏取材，进行系列检测。组织病理学石蜡切片苏木素-伊红（HE）染色检测心肌炎症分级，天狼猩红染色检测心肌胶原纤维含量，免疫组织化学染色检测心肌巨噬细胞和T细胞浸润，实时定量PCR检测MMP-2、MMP-9的mRNA表达水平，冰冻切片用于原位明胶酶谱法检测明胶酶活性。结果 早期干预组和中期干预组的治疗组大鼠心肌组织石蜡切片显示心肌组织炎症积分均显著低于其相应对照组[分别为1．51±0．36比3．03±1．35（P〈0．05）和2．11±0．82比3．75±0．29（P〈0．01）]，免疫组织化学显示心肌巨噬细胞和T淋巴细胞浸润数目均显著低于其相应对照组，天狼猩红染色显示心肌间质纤维积分和纤维含量均明显低于其相应对照组[分别为1．51±0．35比2．75±0．29（P〈0．01）和1．61±0．42比2．50±0．41（P〈0．05）]，实时定量PCR检测心肌MMP-Objective To investigate the effects of matrix metalloproteinase-9 （MMP-9） inhibitor minocyclin hydrochloride in Lewis rats with experimental autoimmune myocarditis （EAM）. Methods EAM was induced by injection of cardiac C protein emulsified in completed Freund adjuvant in double footpad and intraperitoneal injection of pertussis toxin on 6-to 8-week old Lewis rats. Sixty EAM Lewis rats were divided into 3 groups （ early, middle and late intervention groups, n = 20 each ： 10 minocyclin treated and 10 control rats）. In early intervention group, rats in treatment group received intraperitoneal injection of minocyclin hydrochloride from 1 ^st to 21^st day after immunization; in middle intervention group, rats were treated from 8^th to 28^th day after immunization and in late intervention group, rats were treated from 15^th to 35^th day after immunization （50 mg/kg body weight, once daily）. Control rats received iutraperitoneal injection of same volumetric physiological saline at corresponding time periods. At the end of intervention, rats were euthanatized and hearts were harvested. Paraffin sections were used for hematoxylin and eosin stain to determine the inflammatory score, for picrosirius stain to determine fibrosis score and collagen content, and for immunohistologieal stain to determine macrophages and T lymphocytes. Real time PCR was used to detect mRNA expression of myocardial MMP-2 and MMP-9. Cryostat sections were used for in situ zymography to detect protein activity of gelatinase. Results Inflammatory score in cardiac paraffin slides, number of cardiac macrophages and T lymphocytes, cardiac interstitial fibrosis score and content, expression of MMP-2, 9 mRNA and activity of gelatinase in treatment group were all significantly lower than in control group for early and middle intervention groups （ inflammatory score ： early control group vs. treatment group ： 3.03± 1.35 vs 1.51 ± 0. 36,P 〈 0. 05, middle control group vs. treatment group ： 3. 75 ± 0. 29 vs. 2. 11 ± 0. 82,P �

关 键 词：心肌炎 MMP-9 米诺环素 

分 类 号：R965[医药卫生—药理学]