非酒精性脂肪肝（NAFLD）的发病率越来越高，已经成为许多国家慢性肝病的主要病种，单纯性脂肪肝引发肝细胞癌的病例数量也在不断上升。近期研究发现高脂饮食诱发的肠道菌群结构失衡在NAFLD等代谢性疾病的发生发展过程中起着重要作用。并且目前还缺乏针对NAFLD的有效治疗方法。本项目组研究提示益气清化方在改善脂质代谢和减轻肝细胞炎症的同时，也具有改善NAFLD动物模型菌群紊乱的作用。项目组拟运用高脂饮食建立非酒精性脂肪肝大鼠模型，通过观察肠道菌群紊乱、及伴随的门静脉内毒素升高、Th17/Treg细胞失衡和其相关的细胞因子网络变化等，从组织学、微生态学和细胞分子生物学等方面研究NAFLD的发病机制，并研究益气清化方干预的主要作用靶点，为中医药治疗和预防NAFLD提出新的依据和方法。

With the increasing incidence of NAFLD has become a major cause of chronic liver disease in many countries.Particular concern in recent years, simple fatty degeneration caused by the number of cases of HCC are rising. Recent studies have found that high-fat diet-induced intestinal microflora imbalance plays an important role in the occurrence of NAFLD and metabolic diseases. However, there is lack of effective treatment for NAFLD at present. The experiment by the Chinese intervention on non-alcoholic fatty liver disease in rats, by observing the blank group, the control group rats, the different structure of the gut microbia, serum ALT, gamma-GT, TC, TG and serum drug intervention change of toxins and liver pathology to study Chinese method of Yiqi-Qinghua through the regulation of intestinal flora disorders to improve non-alcoholic fatty liver disease in rats. Therefore, the research group intends to establish the use of high-fat diet model of non-alcoholic fatty liver in rats, by observing the proportion of cells in the intestinal flora, serum endotoxin, and Th17/Treg and associated changes in cytokine network imbalance; from histology, micro-aspects of cell and molecular biology to study NAFLD, such as ecology and pathogenesis of NAFLD. And study of traditional Chinese medicine Yiqi Qinghua Fang’s main targets of intervention for the treatment and prevention of NAFLD with TCM to propose new experimental evidence and methods.

非酒精性脂肪性肝病（non-alcoholic fatty liver disease, NAFLD） 是包括单纯性脂肪肝到脂肪性肝硬化的一组疾病谱，已经成为许多国家慢性肝病的主要病因。近年来大量研究表明，高脂饮食诱导的Th17/Treg细胞失衡及肠道菌群结构紊乱在 NAFLD 等代谢性疾病的发生过程中起着重要作用。本研究观察中药复方在治疗高脂饮食诱导的NAFLD大鼠模型过程中对Th17/Treg细胞失衡及肠道菌群的调节作用。研究将60只雄性Wistar大鼠分为空白组、黄连素对照组及低、中、高中药组，每组10只。空白组给予普通饲料，模型组、对照组及各中药组给予高脂饲料。中药组给予中药复方药液高、中、低剂量灌胃，对照组给予黄连素液灌胃，模型组和空白组灌服等量的0.9%NaCl溶液，各组均干预12周。实验结束后，采用流式细胞术检测大鼠外周血及肠道组织的Th17/Treg细胞比例，通过对16S rDNA的变性梯度凝胶电泳测序技术和ERIC-PCR指纹图谱检测大鼠肠道菌群结构的变化，并且测定大鼠血清肝功能（ALT、AST、GGT、TG、TC），血浆内毒素（LPS）及相关细胞因子（TGF-β、TNF-α、IL-6、IL-8、IL-10、IL-17）水平；采集大鼠肝组织，HE染色后观察肝脂肪变性情况。NAFLD大鼠外周血Th17细胞比率及相关细胞因子（IL-17、IL-6、IL-8）较空白组明显增高（P<0.05）, 中药高剂量组、中剂量组及对照组较模型组显著降低（P<0.05）。模型组NAFLD大鼠肠道固有层Treg细胞比例显著低于空白组(P<0.05)；模型组大鼠肠道固有层Th17/Treg细胞比例高于空白组（P<0.05）。模型组较空白组肠道菌群数目及丰富度明显降低，各中药治疗组大鼠均较模型组肠道菌丰富度更高，而与空白组相似；在门水平上，空白组厚壁菌门数低于模型组；空白组大鼠肠道菌群中的Flintibacter butyricus数量高于模型组，经中药治疗后数量则明显升高。本研究结果显示，NAFLD大鼠的外周血中Th17/Treg比例及相关的细胞因子水平高于空白组，并且存在肠道菌群紊乱。中药复方干预可能通过调节Th17/Treg失衡、纠正肠道菌群紊乱，从而减轻NAFLD大鼠肝细胞的炎症和肝脏的脂肪沉积。