摘要
目的:观察佐剂性关节炎(Adjuvant Arthritis,AA)大鼠心功能、MMP-9及TIMP-1蛋白在心肌中表达。方法:将24只大鼠随机分为正常对照组(Normal Control,NC)、模型对照组(Model Control,MC),每组12只,分别向模型对照组动物右后足跖皮内注射弗氏完全佐剂(Freund's Adjuvant Complete,CFA)致炎,致炎48天后,观察两组大鼠足跖肿胀度及关节炎指数(Arthritis Index,AI),分别通过小动物多道生理记录仪和超声心动图检测心功能,HE染色观察心脏组织病理学改变,免疫组化染色法检测基质金属蛋白酶9(Matrix Metallo Proteinase 9,MMP-9)、基质金属蛋白酶抑制因子-1(Tissue Inhibitor of MetalloProteinase-1,TIMP-1)的蛋白表达情况,ELISA法测定血清细胞因子的变化,流式细胞仪测定调节性T细胞(Regulation T cell,Treg)的表达。结果:①与NC组比较,MC组大鼠致炎12天体重出现下降,于致炎48天体重下降至最高峰;致炎18天足跖肿胀度达最高峰。②与NC组比较,MC组的舒张早期血流峰值速度(early diastolic peak flow velocity,E)、E/A、短轴缩短分数(left ventricular fraction shortening,FS%)显著降低(P<0.05或P<0.01),舒张晚期血流峰值速度(late diastolic peak flow ve-locity,A)显著升高(P<0.01)。与NC组比较,MC组心率(heart rate,HR)、心脏质量指数(heart index,HI)、左室收缩末压(Left Ventricular Systolic Pressure,LVSP)、左室舒张末压(Left Ventricular End-diastolic Pressure,LVEDP)显著升高(P<0.05),左室内压上升/下降最大速率(Maximum Rate of Ventricular Pressure of development or decline,±dp/dtmax)显著降低(P<0.05)。③与NC组比较,MC组TNF-α、BNP、IL-17显著升高(P<0.05或P<0.01),IL-10、CD4+Treg、CD4+CD25+Treg显著降低(P<0.01)。④与NC组比较,MC横截面心肌细胞横径值(TDM/μm)显著升高(P<0.05),MMP-9表达量在MC组大鼠心肌细胞显著升高(P<0.01);TIMP-1的表达量则显著降低(P<0.01)。⑤Spearman相关性分析显示:心功能参数E峰与TIMP-1蛋白染色指数呈正相关;A峰与TIMP-1蛋白染色指数呈正相关,与MMP-9呈负相关;心脏质量指数(HI)与关节炎指数(AI)呈正相关;+dp/dtmax与IL-17呈显著负相关,与TIMP-1蛋白染色指数呈正相关;-dp/dtmax与IL-10呈负相关(P<0.05)。结论:AA大鼠存在心功能下降。其机制可能是大鼠在致炎后对抗原刺激呈高敏反应状态,免疫调节功能紊乱,释放大量细胞因子和炎症介质,导致局部关节的病变的同时,心肌细胞及细胞外基质亦发生损害,从而发生AA心功能降低。
Objective：To observe the cardiac function,MMP9 and TIMP1 protein expression in myocardium in adjuvant arthritis rats.Methods：24 rats were randomly divided into control group（NC） and model control group（MC）.Except for control group,the rats of model control group were induced to AA model by intradermally injecting Freund′s complete adjuvant into the right paw.48 days after inducing AA model,the swelling degree of voix pedis and arthritis index（AI） were observed in rats.Cardiac function was detected by the small animal multi-channel physiological recorder and heart echocardiography.Heart tissue pathological were observed by HE staining and matrix metalloproteinase-9（Matrix metallo proteinase 9,MMP-9）,matrix metalloproteinase inhibitor-1（Tissue inhibitor of metallo proteinase-1,TIMP-1） expression of the protein were detected by immunohistochemistry staining in AA rats.Serum cytokines were measured by ELISA method and the expression of regulation T cells（Treg） were observed by flow cytometry.Results：①Compared with those in NC group,MC group declined inflammatory 12 d weight,weight loss on inflammatory 48 d to peak;18 d inflamed paw swelling reached its peak.② Compared with those in NC group,E peak,E/A,FS% of MC group were significantly lower（P0.05 or P0.01）,A peak was significantly higher（P0.01）.Compared with those in NC group,HR,HI,left ventricular end systolic pressure（LVSP）,left ventricular end diastolic pressure（LVEDP） of MC group increased significantly（P0.05）,left ventricular pressure rise / fall maximum rate（± dp/dtmax） were significantly lower（P0.05）.③ Compared with those in NC group,TNF-α,BNP,IL-17 of MC group were significantly higher（P0.05 or P0.01）,IL-10,CD4＋ Treg,CD4＋ CD25＋ Treg were significantly lower（P0.01）.④ Compared with those in NC group,cross-sectional myocardial cell diameter values（TDM/μm） of MC group was significantly higher（P0.05）,MMP-9 expression in MC was significantly higher in myocardial cells,significant difference（P 0.01）,while TIMP-1 expression level was significantly lower（P0.01）.⑤ Spearman correlation analysis showed that,cardiac function in AA rats with peak E and TIMP-1,peak A and TIMP-1,HI and AI,﹢dp/dtmax and TIMP-1 were positively correlated（P0.05）.Peak A and MMP-9,＋dp/dtmax and IL-17,-dp/dtmax and IL-10 showed a significant negative correlation（P0.05）.Conclusion：The cardiac function of AA rats was reduced.The mechanism may be results from inflammation of AA rats which showed a high sensitive response to antigen stimulation and disorder of immune regulation,released of a large number of cytokines and inflammatory mediators,leaded to damage of local joints as well as cardiac cells and extracellular matrix.Finally,cardiac function was reduced in AA rats.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2012年第3期251-256,265,共7页
Chinese Journal of Immunology
基金
国家中医药重点学科中医痹病学建设项目(国中医药发[2009]30号)
安徽省科技厅科研计划项目(No.11010402170)
安徽中医内科应用基础与开发研究省级实验室项目(科条[2008]150号)
