基于NF-κB信号转导通路探讨丹蒌方治疗冠心病痰瘀互结证的机制研究

Coronary heart disease can be ascribed to category of pectoral stuffiness pain, precordial pain, according to clinical manifestation. There is more the combined Zhenghou than simple Zhenghou of coronary heart disease in clinical practice. Turbid phlegm, blood stasis, blending of turbid phlegm and blood stasis syndrome is most common Zhenghou of coronary heart disease. The prophase study of literature indicated that NF-κB is the important target point in the NF-κB signal pathway. NF-κB signal pathway is throughout the pathological process of lipid disorders and microcirculation disturbance, which are closely related to coronary heart disease with the blending of turbid phlegm and blood stasis syndrome. The prophase study results suggested that ApoE and eNOS single or double-knockout mouse model could be used to simulate different state of turbid phlegm, blood stasis as well as blending of turbid phlegm and blood stasis of coronary heart disease. So, in the study, our group would reveal the main regulation mechanism and target point of coronary heart disease with the blending of turbid phlegm and blood stasis syndrome based on NF-κB signal pathway and endocrine system by the ApoE and eNOS single or double-knockout mouse model and detect the NF-κB correlated modulin, the level of inflammatory factor, matrix metalloproteinases and thyroid hormone by the methods of ELISA, histopathologic slide and so on. And then investigate the mechanism of coronary heart disease with the blending of turbid phlegm and blood stasis syndrome and provide the experimental evidence for coronary heart disease with the blending of turbid phlegm and blood stasis syndrome treated with Danlou Fang.

冠心病根据其临床表现可归于中医学胸痹、心痛范畴。冠心病单纯证候较少，临床上以复合证候为主，其中痰浊、血瘀、痰瘀互结证为冠心病的主要证候类型。前期文献研究表明NF-κB是NF-κB信号转导通路的重要靶点，而NF-κB信号转导通路的调控作用又贯穿于与冠心病痰瘀互结证密切相关的脂代谢紊乱、微循环障碍的整个病理过程。本课题组前期实验初步建立的ApoE和eNOS单敲/双敲小鼠模型能够模拟冠心病痰浊、血瘀、痰瘀互结证的不同状态，因此本研究拟利用ApoE和eNOS单敲/双敲小鼠模型，通过ELISA、病理组织切片等方法，检测NF-κB相关调控蛋白、炎症因子、基质金属蛋白酶以及甲状腺激素的水平，从NF-κB信号转导通路以及内分泌系统等多角度，揭示其对冠心病痰瘀互结证的主要作用靶点，探讨丹蒌方调治冠心病痰瘀互结证的作用机制，为丹蒌方治疗冠心病痰瘀互结证提供实验依据。

冠心病根据其临床表现可归于中医学胸痹、心痛范畴。冠心病单纯证候较少，临床上以复合证候为主，其中痰浊、血瘀、痰瘀互结证为冠心病的主要证候类型。前期文献研究表明NF-κB是NF-κB信号转导通路的重要靶点，而NF-κB信号转导通路的调控作用又贯穿于与冠心病痰瘀互结证密切相关的脂代谢紊乱、微循环障碍的整个病理过程。本研究利用喂养高脂饲料的ApoE基因敲除小鼠模拟“痰浊”，利用注射异丙肾上腺素的ApoE基因敲除小鼠模拟“血瘀”，利用给ApoE基因敲除小鼠喂养高脂并注射异丙肾上腺素的小鼠模拟“痰瘀互结”，通过观察小鼠的一般状态、检测小鼠血脂水平，运用小动物超声检测小鼠左心室功能共同评价小鼠模型的建立方法，结果表明此模型建立方法可以用于课题组后续的药物干预实验和机制研究。在研究过程中，通过Western Blot、ELISA、病理组织切片等方法，检测了NF-κB相关调控蛋白、炎症因子、基质金属蛋白酶以及甲状腺激素的水平，从NF-κB信号转导通路以及内分泌系统等多角度，初步揭示了其对冠心病痰瘀互结证的主要作用靶点，探讨了丹蒌方治疗冠心病痰瘀互结证的作用机制。. 该项目主要创新点是利用ApoE基因敲除小鼠通过高脂饲料喂养结合异丙肾上腺素诱导建立了冠心病痰瘀互结证小鼠模型的基础上，模拟了小鼠痰浊、血瘀、痰瘀互结的3种不同状态，从NF-κB信号转导通路和内分泌系统等多角度初步揭示了丹蒌方治疗冠心病痰瘀互结证的作用靶点和机制，为丹蒌方治疗冠心病痰瘀互结证提供实验依据。