基于代谢性核受体的代谢综合征痰证分子机制的研究

Metabolic nuclear receptors (MNRs) are a group of nuclear hormone receptor transcription factors, which are very closely related with metabolic syndrome (MS) including insulin resistance. Our preliminary studies showed that phlegm was the main traditional Chinese medicine (TCM) pathological factor of MS and an important factor for causing MS insulin resistance, indicating that there exists certain relevance between phlegm syndrome and MNRs. Therefore, this subject will illustrate the intrinsic relations between phlegm syndrome of MS and metabolic nuclear receptors based on the theory of correspondence between syndrome and prescription. First, this project will set the compound data of clinical common prescription of clearing phlegm for MS, which collected from the chemical structure knowledgebase of TCM constructed by our former work and MNRs (PPARs, LXRs, FXRs and RXRs) as the study subjects. And then, we will establish MNRs activity predicted models to screen molecular targets of MS phlegm syndrome preliminarily by computer virtual screening methods. And then, we will adopt wendan tang to interven the animal model of MS phlegm syndrome and detect the molecular targets expressions of MS phlegm syndrome in order to find the final targets. Finally, we will search for action pathway of those targets and make biological functions notes from genetic databases to construct collaborative networks of phlegm syndrome target functions by use of network clustering analysis technology. In summary, this project will be hoped to discover the multidimensional molecular mechanism of MS phlegm syndrome from the levels of computer molecular simulation, protein and cell, and also to provide some molecular basis for MS phlegm syndrome.

代谢性核受体(MNRs)是一组与代谢调节相关的配体激活核受体转录因子，与代谢综合征(MS)包括胰岛素抵抗等密切相关。课题组前期研究显示痰是MS最主要的中医病理因素，是导致MS胰岛素抵抗的重要病因。故MS痰证与MNRs存在一定的相关性。因此，为了阐明二者的内在关系，本课题基于方证相应理论，在前期建设中医药信息数据库基础上，先以MS临床常用化痰基础方温胆汤所含化合物数据和MNRs (PPARs, LXRs, FXRs和RXRs) 为研究对象，通过计算机分子筛选方法建立MNRs活性预测模型，初步筛选MS痰证分子靶标；再通过温胆汤干预MS痰证动物模型，用免疫组化对计算机初筛的分子靶标表达情况进行检测，确定MS痰证分子靶标；最后通过对MS痰证分子靶标的作用通路进行检索、生物功能注释和网络聚类，构建MS痰证分子靶标协同网络通路，从不同角度探讨MS痰证的多维调控分子机制，为MS痰证诊断提供部分分子基础。

代谢综合征(metabolic syndrome, MS)是以中心性肥胖、糖调节受损、血脂异常等多重危险因素在个体集聚的复杂症候群，主要临床后果为心脑血管疾病和2型糖尿病，严重危害健康。痰可引起人体各脏腑系统功能失常，而MS的病变涉及各脏腑系统，故痰是引起MS发病的重要原因。课题组前期研究显示痰是MS最主要的中医病理因素，是导致MS胰岛素抵抗的重要病因。代谢性核受体(MNRs)是一组与代谢调节相关的配体激活核受体转录因子，与MS包括胰岛素抵抗等密切相关。故MS痰证与MNRs存在一定的相关性。因此，为了阐明二者的内在关系，本课题在前期建设中医药信息数据库基础上，先以MS临床常用化痰基础方温胆汤所含化合物数据和MNRs (PPARs, LXRs, FXRs和RXRs) 为研究对象，通过计算机分子筛选方法建立MNRs活性预测模型，初步筛选MS痰证分子靶标；再通过温胆汤干预MS痰证动物模型，用免疫组化对计算机初筛的分子靶标表达情况进行检测，确定MS痰证分子靶标；最后通过对MS痰证分子靶标的作用通路进行检索、生物功能注释和网络聚类，构建MS痰证分子靶标协同网络通路，探讨MS痰证的调控分子机制。计算机分子筛选研究结果研究表明MS痰证与PPARα, PPARβ, PPARγ, LXRα ,LXRβ和RXRα六个靶标密切相关；中药-靶标-生物通路网络分析显示，19条生物通路在WDD干预MS痰证中受到了影响，WDD的主要药理作用涉及到维持脂质和葡萄糖代谢和抗癌活性以及免疫调节和保肝作用,说明MS痰证涉及到脂质和葡萄糖代谢异常及免疫失调且可致癌和肝脏病变。动物实验研究结果表明高糖高脂可以建立大鼠MS痰证模型，温胆汤能改善MS大鼠肥胖指标、改善体内血脂及胰岛素抵抗的作用，且与二甲双胍相比，温胆汤干预作用更显著，为临床从痰论治MS奠定实验基础。MS痰证大鼠内脏HE染色实验结果表明，痰致病广泛，在肝脏、腹部脂肪等器官组织均可致病。免疫组化实验结果表明MS痰证大鼠肝脏的靶标受体表达受到影响，与计算机分子筛选研究部分结果相互印证，说明计算机分子筛选研究结果可以为动物实验研究提供指导，对中医药现代化研究具有重要意义。