基于网络药理学清热燥湿中药调节脂质代谢药效物质及作用机制研究

Although alleviates fever eliminating dampness Traditional Chinese Medicines contain various type of natural products, they showed almost the same improve effects on abnormal lipid metabolism in traditional clinic. We suggested that different constituents of alleviates fever eliminating dampness Traditional Chinese Medicines act at different key target of lipid metabolism network to display intervention lipid metabolism through a multi-direction pattern. In this project, Five kind of most common used fever eliminating dampness Traditional Chinese Medicines, Scutellaria baicalensis, Coptis chinensis, Phellodendron chinense, Gentiana scabra and Sophora flavescens were selected as research objects, serum containing herb medicines will be administrated to oleic acid induced triglyceride accumulation liver cell lines, and then gene chip and antibody chip will be used to find the gene and protein expression difference, further, using a network pharmacology method, a multi-level structural network analysis method will be established to evaluate total effectiveness and relative relationship between targets of metabolism regulation effect, the key node and functional module will be confirmed. Next, combined with phytochemistry research result, using virtual screening and molecular docking method to predict pre-active constituent, and the regulation effects on lipid metabolism will be validated in vivo and in vitro. Finally, the mechanism and active compounds of fever eliminating dampness Traditional Chinese Medicines will be partly clarified and the common rule will be summarized. From above research, the new target for lipid metabolism regulation is hopeful to be found. The research results can also provide scientific evidence to Traditional Chinese Medicine clinical rational administration and new drug development.

清热燥湿中药化学成分结构差异较大,但临床改善脂质代谢异常作用类似,推测清热燥湿中药不同化学成分分别作用于脂质代谢网络关键靶点，通过多向调节方式形成类似的干预脂质代谢效果。本项目拟选取黄芩、黄连、黄柏、龙胆、苦参等五种临床常用调节脂质代谢异常清热燥湿中药，制备含药血清,对油酸诱导脂肪蓄积肝细胞模型给药,采用基因芯片和抗体芯片技术比较基因和蛋白的表达差异；采用网络药理学方法，建立多层次结构化网络分析方法，评价清热燥湿中药调节脂质代谢的整体影响以及靶点间相互作用关系,确定关键节点和功能模块；结合化学成分研究结果,采用分子对接和虚拟筛选方法，预测可能药效物质；开展体内、外活性验证，明确清热中药改善脂质代谢异常药效物质物质及作用机制，总结规律，以期从中药临床经验中发现改善脂质代谢异常新靶点，为指导中药临床合理用药及创新药物发现提供科学依据。

本项目根据申请书假说，推测清热燥湿所含中药化学成分虽然不同，但临床改善脂质代谢异常作用类似，不同化学成分分别作用于脂质代谢网络关键靶点，通过多向调节方式形成类似的干预脂质代谢效果。.本项目选取临床常用调节脂质代谢异常清热燥湿中药，制备含药血清,对油酸诱导脂肪蓄积肝细胞模型给药,采用基因芯片和抗体芯片技术比较基因和蛋白的表达差异；采用网络药理学方法，建立多层次结构化网络分析方法，评价了清热燥湿中药调节脂质代谢的整体影响以及靶点间相互作用关系，结合文献报道，明确了以AMPK信号通路是清热药调节脂代谢异常的共性通路, 发表ESI高被引论文1篇；重点对文献报道对AMPK具有激活作用的化学成分进行了虚拟筛选结合实验验证研究。.采用乳清酸诱导脂质代谢异常大鼠以及KKAy小鼠，以AMPK为核心开展了黄连、黄柏、龙胆、苦参调节脂代谢异常作用机制研究，并详述了黄芩的研究结果。发现这些清热药水提取物均能上调p-AMPK的水平，促进其下游的p-SREBP-1c和p-ACC的表达，并且能够下调SREBP-1c活性蛋白形式的表达，抑制脂肪酸合成。黄芩中主要成分黄芩苷能调节乳清酸诱导的NAFLD大鼠肝脏中FFA代谢，上调p-AMPK、p-SREBP-1c和p-ACC的表达，对SREBP-1c活性蛋白形式的表达有下调作用，黄芩苷还能下调与FFA合成相关的ACC、FAS、SCD基因转录和蛋白翻译的水平，从而抑制FFA的合成；同时还能促进与脂肪酸氧化相关的CPT-1与Nrf2基因表达，从而促进FFA的氧化，改善FFA水平异常升高引起的肝脂质沉积。进一步采用细胞水平研究明确了其作用机制。.结合本项目研究与项目组前期研究基础，归纳分析了清热药改善肝脏脂质代谢紊乱的主要机制，总结出：清热燥湿中药可通过激活AMPK，增加脂肪酸氧化、减少甘油三酯的合成而发挥调节脂质代谢作用。.通过对清热药调控脂质代谢的研究，部分阐明了中药临床治疗代谢性疾病的基本规律，为中医临床干预糖脂代谢辨证用药提供了科学依据。.本研究在国内外学术刊物上发表论文 6 篇，其中 SCI 收录论文 5 篇；培养研究生 5 名, 申请专利 2项。.