基于LXR-AVP/RAAS复杂式调控通路的慢性心力衰竭“温阳消饮”法多靶点疗效作用机制研究

Abstract：Chronic heart failure（CHF）is classified as Traditional Chinese medicine（TCM） phlegm Category.Numerous clinical and experimental studies confirm: Wen Yang Xiao Yin therapy is the classic effective treatment method of the syndrome,and the mechanism is Worthy of further exploration. Supported by the the early Natural Science Foundation Project findings, Our group found that Lingguizhugan and Tinglidazaoxiefei decoction,as the representative prescription of the therapies,can significantly improve the CHF rats excessive activation of RAS system, cardiac remodeling ,and abnormal expression of aquaporin in lung, kidney and other tissues,and the microarray shows the differentially expressed genes mainly related adipokines pathways. As the transcription factor, LXR is the downstream regulatory targets of the adipocytes pathway, Regulating the body's fluid balance by the LXR-AVP-AQPs pathway,and multi-target effecting on the RAAS system, and ultimately improve myocardial remodeling. Based on this, under the guidance of the TCM phlegm theory,the research will use an entry point that transcription factor LXR regulate gene expression in space-time polymorphism, and be carried out closely around the hypothesis "LXR-AVP / RAAS complex pathways involved in the regulation of CHF mouse liquid water balance and neuroendocrine mechanisms to improve cardiac remodeling is the critical core mechanism of Wenyangxiaoyin treatment ‘s multi-target effect on CHF".Intending to adopt FXR knockout mice, ELISA and other biological techniques,the research explore the critical core mechanism of Wenyangxiaoyin treatment ‘s multi-target effect on CHF, provide the scientific basis for experimental basis To illustrate the theory of phlegm and Wenyangxiaoyin treatment.

慢性心力衰竭（CHF）属中医痰饮范畴，温阳消饮法是该病证经典有效治法，其作用机制值得进一步探索。前期自然基金课题研究发现：温阳消阴法可显著改善CHF大鼠RAAS系统过度激活、心肌重塑及肺、肾组织多种水通道蛋白表达异常，基因芯片检测示该治法差异表达基因主要与脂肪细胞因子通路等相关。转录因子LXR作为脂肪细胞因子通路下游调节靶点，通过LXR-AVP-AQPs调节机体水液平衡，并对RAAS系统有多靶点效应，最终改善心肌重塑。本课题在痰饮理论认识指导下，以转录因子LXR调控基因表达时空多态性为切入点，密切围绕“LXR-AVP/RAAS复杂式调控通路参与调节CHF小鼠水液平衡和神经内分泌机制从而改善心肌重塑是温阳消饮法治疗CHF多靶点疗效的关键核心机制”假说开展研究。拟采用LXR敲除小鼠、EMSA等生物学技术，探索该法的CHF多靶点疗效关键核心机制，为阐释痰饮理论及温阳消饮治法的科学基础提供实验依据

慢性心力衰竭（CHF）属中医痰饮范畴，温阳消饮法是该病证经典有效治法。课题组研究发现：温阳消饮法可显著改善CHF大鼠RAAS系统过度激活、心肌重塑及肺、肾组织多种水通道蛋白表达异常，基因芯片检测提示该治法差异表达基因 主要与脂肪细胞因子通路等相关。本研究在痰饮理论认识指导下，以转录因子LXR调控基因表达时空多态性为切入点，研究温阳消饮法基于LXR-AVP/RAAS/NF-κB复杂式调控通路调节CHF小鼠水液平衡和神经内分泌失调从而治疗CHF多靶点疗效机制。研究首先成功复制阿霉素诱导CHF小鼠模型，实施温阳消饮法等相应干预，研究结果证实在小鼠心脏组织温阳消饮法可改善心肌细胞凋亡、心室重构，其作用机制与下调LXRαmRNA以及蛋白水平表达，进而致NF-κB及下游调节因子iNOS和TNF-α表达水平上调有关；温阳消饮法对小鼠血清RAAS系统水平和AVP水平均有下调作用，在下丘脑和肾脏组织下调LXRβmRNA和蛋白水平表达，上调肾脏组织AQP2mRNA和蛋白水平表达，通过对RAAS系统和AVP-AQP2 神经内分泌激素调节改善CHF小鼠水液代谢异常。温阳消饮法在心脏可改善心脏细胞代谢和心功能，体现“温阳”之功，同时也可促进水液代谢，发挥“利水”之效，具有多靶点疗效，且温阳消饮法中“茯苓-桂枝”药对具有双向调节作用，体现了痰饮治疗的原则“和之”，丰富了“温药和之”的又一层含义，根植于中医治疗追求以平为期的人本思想。研究结果为阐释中医经典痰饮理论及代表治法温阳消饮法的科学内涵的创新性阐释。