基于Chymase介导非ACE依赖的AngII生成途径探研冠心V号干预心室重构机制

GuanXin Ⅴ Formula (a hospital preparation) is effective in treating coronary atherosclerotic heartdisease(CAD).Our team had revealed that the formula may improve cardiac function and decelerate ventricular remodeling by inhibiting inflame-immune reaction,down-regulating expression of AT1R and up-regulating AT2R in post-MI model of Rats.，which is related to RAAS and differ from ARBs.As the latest research indicates one of the key factors of Postinfarct ventricular remodeling is the mast cell and its bioactive molecule Chymase, which has crosstalk with the RAAS.Wehypothesize that GuanXin Ⅴ Formula decelerate the remodeling by Chymase way. The main experimental contains in vivo and in vitro. The former includes making models of postinfarct left ventricular remodeling(induced by left anterior descending coronary artery ligation) in hamsters, administering these animals with GuanXin ⅤFormula and other contrast medicine, observation the activity of Chymase-related ACE-independent AngII production way. The latter includes cultivation primary hamster peritoneal mast cells , cardiac fibroblasts and myocardial cells, activation the cells with C48/80 or purified Chymase, observation the effect of serum ingredients of GuanXin Ⅴ Formula by molecularbiologic and immunologic techniques, such as Western blotting, siRNA and immunohistochemistry. In a word, the study will elucidate the key mechanism of the formula on the postinfarct left ventricular remodeling at cellular and molecular level.

冠心Ⅴ号（GXⅤ）是临床治疗冠心病的有效中药复方制剂（医院制剂），可抑制冠心病患者及心肌梗死（MI）模型大鼠的炎症免疫反应，降低AT1R及增加AT2R的表达，延缓心室重构（VR）、改善心功能。前期研究发现GXⅤ干预VR的机制与RAAS密切相关但又不同于ARBs。关键蛋白酶Chymase通过转化生成AngⅡ、促心肌纤维化及心肌细胞凋亡等机制促进VR，是与RAAS有交互作用的重要因子。故课题组提出GXⅤ通过Chymase介导途径干预VR的假说。设计整体动物及体外细胞实验以验证假说：拟观察GXⅤ干预MI模型仓鼠VR，从Chymase介导非ACE依赖AngⅡ生成途径探研其关键药效机制；用GXⅤ含药血清预处理仓鼠腹腔肥大细胞、心肌成纤维细胞、心肌细胞，观察C48/80、Chymase诱导或干预上述细胞后的效应。涉及siRNA、IHC、Western blot、流式细胞技术等分子生物、免疫学方法。

冠心Ⅴ号（GXⅤ）是临床治疗冠心病的有效中药复方制剂（医院制剂），可抑制冠心病患者及心肌梗死（MI）模型大鼠的炎症免疫反应，降低AT1R及增加AT2R的表达，阻遏心室重构（VR）、改善心功能。前期研究发现GXⅤ干预VR的机制与RAAS和氧化应激等密切相关，本研究围绕此关键环节展开研究以探讨GXV阻遏甚至逆转MI后VR的生物学机制。我们发现GXV可以通过Chymase介导非ACE依赖Ang Ⅱ生成途径并通过抑制TGF-β信号通路来抑制RAAS系统激活和肥大细胞聚集，抑制心肌细胞凋亡、纤维化和心室重构，同时这一过程中的TGF-β1关键分子与Vimentin存在直接互作关系。