柴胡人参药对上调FXR表达调节胆汁酸代谢及肠道菌群分布 防治非酒精性脂肪性肝病的机制研究

Based on the results of our recent studies, the hypothesis was made that Drug Pair of Ginseng and Radix bupleuri (DPGR) on behalf of soothing liver and invigorating spleen recipe（SLISR） might be used to treat NAFLD due to regulating bile acid metabolism disorders and constitute of intestinal flora by farnesoid X receptor (FXR) pathway. NAFLD rat model is prepared by high lipid and sugar recipe. The histology and biochemical indexes of rats can be used for the general assessment of the therapeutic effect of DPGR. Immunofluorescence, Laser confocal scaning microscopy(LCSM), Real-time quantitative PCR and Western Blot analytic techniques are adopted to observe bile acid metabolism regulation of FXR mediated pathway, combined with the whole genome sequencing methods to detect the constitute of intestinal flora of NAFLD rat. Through this topic, we try to reveal the relationship of those three factors about “farnesoid X receptor—bile acid metabolism—intestinal flora” Under the intervention of DPGR, and then to elucidate the action and therapeutic targets for its treatment and research the modern medical mechanism of SLISR for NAFLD. So that will provide new ideas for prevention and treatment of NAFLD by traditional Chinese medicine.

基于我们前期的工作基础，提出“代表疏肝健脾法的柴胡人参药对治疗NAFLD的主要机制可能是通过法尼醇X受体（FXR）途径调控胆汁酸代谢紊乱及肠道菌群构成”的假说。高脂高糖配方制备NAFLD大鼠模型，利用普通病理学和生化学方法综合评判柴胡人参药对治疗大鼠NAFLD的疗效；采用免疫荧光和激光共聚焦检测、实时荧光定量PCR及Western Blot分析，观察药对对FXR介导的胆汁酸代谢调节通路的影响，结合全基因组重测序方法检测NAFLD大鼠肠道菌群构成，揭示柴胡人参药对干预下“法尼醇X受体—胆汁酸代谢—肠道菌群”三者之间的关系，阐明其治疗NAFLD的作用机制和治疗靶点，从实验医学角度研究疏肝健脾法治疗NAFLD的现代医学机理。为中医药防治NAFLD提供新思路。

基于我们前期的工作基础，提出“具有疏肝健脾功能的柴胡人参药对治疗NAFLD的主要机制可能是通过法尼醇X受体（FXR）途径调控胆汁酸代谢紊乱及肠道菌群构成”的假说。高脂高糖配方制备NAFLD大鼠模型，利用普通病理学和生化学方法综合评判发现柴胡人参药对能够减轻大鼠肝脏脂肪蓄积，减低血清甘油三脂，调节大鼠糖代谢，药物干预后初级胆汁酸CA、GCA、GCDCA、TCA和次级胆汁酸6_ketoLCA、TDCA、muroCA、HDCA、GHDCA、GLCA、TLCA、TUDCA、THDCA、TDCA水平升高；采用实时荧光定量PCR及Western Blot分析，发现柴胡人参药对能够上调FXR及SHP蛋白表达，下调相关下游蛋白表达，从而调节胆汁酸。利用16s rRNA技术分析发现药物干预后大鼠肠道菌群多样性提高，利用功能与通路预测分析发现p108-pwy,pwy-1269,colansyn-pwy,pwy-7315,pwy-6470 pwy-6969三羧酸循环V(2-氧戊二酸:铁氧还蛋白氧化还原酶)功能存在显著差异。提示柴胡人参药对防治非酒精性脂肪性肝病可能与铁氧还蛋白氧化还原酶的活性有关，为后续提供研究方向。