新风胶囊对佐剂性关节炎大鼠心肌纤维化的影响及机制研究

Effects of Xinfeng Capsule on Myocardial Fibrosis in Adjuvant Arthritis Rats and Its Mechanism Research

目的探讨益气健脾通络中药复方新风胶囊对佐剂性关节炎(AA)大鼠心肌纤维化的影响及其作用机制。方法 60只Wistar雄性大鼠随机分为正常对照组(12只)和模型组(48只),除正常对照(NC)组外,每只大鼠右足跖皮内注射弗氏完全佐剂(CFA)0.1mL致炎,制备AA大鼠模型。致炎后第19天将AA模型大鼠组随机分为4组:新风胶囊(XFC)组、甲氨喋呤(MTX)组、雷公藤多甙片(TPT)组和模型对照(MC)组,并分组给药30天后处死,采用透射电镜观察各组大鼠心肌组织超微结构;反转录酶-聚合酶链锁反应(RT-PCR)法及Western blot法测定各组大鼠心肌组织基质金属蛋白酶(MMP9)、基质金属蛋白酶组织抑制因子1(TIMP1)mRNA及蛋白表达水平。结果 (1)电镜观察结果显示,AA大鼠心肌细胞间胶原纤维增生明显,呈现心肌纤维化趋势。MTX、TPT、XFC组均能有效改善心肌超微结构变化,而3组间比较,XFC组大鼠心肌细胞形态完整,细胞核清晰,细胞间仅有少量胶原纤维,明显好于其他两组。(2)与NC组比较,MC组大鼠心肌组织MMP9 mRNA及蛋白表达水平均明显上调,而TIMP1 mRNA和蛋白表达水平明显下调(P<0.01);与MC组比较,XFC组MMP9mRNA及蛋白表达水平均下降,TIMP1 mRNA及蛋白表达水平明显上升,差异有统计学意义(P<0.05,P<0.01)。结论益气健脾通络中药复方新风胶囊可以有效抑制AA大鼠心肌纤维化,其机制可能与降低心肌组织MMP9 mRNA及蛋白表达,提高心肌组织TIMP1 mRNA及蛋白表达水平,调节MMP9/TIMP1平衡,改善心肌细胞外基质代谢有关。

Objective To study effects of Xinfeng Capsule （XFC, a Chinese herbal compound for benefiting qi, invigorating Pi, and dredging collaterals） on myocardial fibrosis in adjuvant arthritis （AA） rats and its mechanism study. Methods Sixty male Wistar rats were randomly divided into the normal control （NC） group （n =12） and the model control （MC） group （n =48）. Except rats in the NC group,Freund′s complete adjuvant （0.1 mL） was intracutaneously injected from the right hind limb to prepare the AA rat model. On the 19th day of inflammation rats in the AA model group were randomly divided into four groups, i.e., the XFC group, the methotrexate （MTX） group, the Tripterygium wilfordii polycoride Tablet （TPT） group, and the MC group,12 in each group. Rats were sacrificed after 30-day medication. The myocardial ultrastructure was observed under transmission electron microscope. The mRNA and protein expression levels of matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （TIMP-1） were detected using RT-PCR and Western blot respectively. Results （1）Results under electron microscope showed obvious hyperplasia of collagen fibers in myocardial cells of AA rats,which indicated the trend of myocardial fibrosis. The myocardial ultrastructural changes could be effectively improved in the MTX group, the TPT group, and the XFC group. The morphology of myocardial cells in the XFC group was contact, with clear nucleus, and little collagen fibers, which was better than the other groups.（2）Compared with the NC group,the mRNA and protein expression levels of MMP-9 in the myocardial tissue of the MC group were obviously up-regulated, while the mRNA and protein expression levels of TIMP-1 were obviously down-regulated （P 0.01）. The mRNA and protein expression levels of MMP-9 decreased, and the mRNA and protein expression levels of TIMP-1 obviously increased in the XFC group, showing statistical difference when compared with the MC group （P 0.05, P 0.01）. Conclusions XFC could effectively inhibit the myocardial fibrosis of AA rats. Its mechanisms might be correlated with inhibiting the mRNA and protein expression levels of MMP-9, improving the mRNA and protein expression levels of TIMP-1, adjusting the balance of MMP-9/TIMP-1,and improving the metabolism of myocardial extracellular matrix.