目的 研究逍遥散对高脂饮食诱导的大鼠代谢相关脂肪性肝病（MAFLD）的保护作用及机制。方法 SD大鼠随机分为5组：对照组、模型组、盐酸吡格列酮（PH，阳性药，5 mg·kg^-1）和逍遥散低、高剂量（19、38 g·kg^-1）组。对照组喂食基础日粮，而其余组喂食高脂肪和高果糖日粮12周，从第13周开始ig给药，每天1次持续4周。试剂盒法检测血清三酰甘油（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、内毒素（LPS）水平，肝组织白细胞介素-6（IL-6）、IL-1β、肿瘤坏死因子-α（TNF-α）水平；HE染色观察肝脏、结肠、空肠和回肠组织病理损伤并评分，油红O染色观察肝脏脂质累积；检测24 h尿液99mTc-喷替酸（DTPA）排泄评价肠黏膜屏障完整性；Western blotting法检测结肠组织toll样受体2（TLR2）、toll样受体4（TLR4）、髓样分化因子88（MyD88）、核因子-κB（NF-κB）、闭锁小带蛋白-1（ZO-1）、封闭蛋白1（Claudin-1）、紧密连接蛋白-1（Occuludin-1）、质膜囊泡相关蛋白1（PV-1）蛋白表达；免疫组化分析结肠和肝组织TLR4、MyD88、NF-κB蛋白表达；免疫荧光检测ZO-1、Claudin-1、E-钙黏蛋白（E-cadherin）蛋白表达。通过16S rRNA基因测序确定肠道菌群种类变化和结构。结果 与模型组相比，逍遥散和PH组血清中TG、TC、LDL-C、ALT、AST和LPS水平显著降低（P＜0.05、0.01、0.001），肝脏IL-1β、IL-6、TNF-α水平显著降低（P＜0.01、0.001），逍遥散高剂量组血清HDL-C水平显著升高（P＜0.01）；逍遥散组肝脏、结肠、空肠、回肠和肝脏病理评分显著降低，肝脏油红O染色区域占比显著降低（P＜0.01、0.001）；免疫组化与Western blotting结果显示，逍遥散和PH组TLR2、TLR4、MyD88、NF-κB蛋白表达显著降低（P＜0.01、0.001）；逍遥散高剂量和PH组大鼠的24 h尿液DTPA排泄显著降低（P＜0.05）；逍遥散高剂量组和PH组ZO-1、Claudin-1和Occludin-1蛋白表达显著升高（P＜0.001），PV-1蛋白表达显著降低（P＜0.001），免疫荧光显示逍遥散高剂量组和PH组的ZO-1、Claudin-1和E-cadherin的表达明显升高。与模型组比较，逍遥散增加肠道菌群丰富度和多样性，减少瘤胃球菌等有害菌、增加拟杆菌等有益菌的相对丰度。结论 逍遥散对高脂饮食诱导的MAFLD大鼠具有改善作用，可通过减轻肝脏炎症、调节肝功能指标，机制可能与修复肠道黏膜屏障、调节肠道菌群、调节TLR4/MyD88/NF-κB通路等相关。

Objective To investigate the protective effects of Xiaoyao San (XYS) on metabolically-related fatty liver disease (MAFLD) induced by high-fat diet in rats and its underlying mechanism. Methods SD rats were randomly divided into five groups: control group, model group, phlorizin (PH, positive drug, 5 mg·kg^-1), XYS low-dose group (19 g·kg^-1), and high-dose group (38 g·kg^-1). The control group was fed a basal diet, while the remaining groups were fed a high-fat and high-fructose diet for 12 weeks. From week 13, the rats were ig administered the drugs once daily for four weeks. Serum triglycerides (TG), total cholesterol (TC), highdensity lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and endotoxin (LPS) levels, and interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) in the liver were determined by kits. The liver, colon, jejunum, and ileum were stained with hematoxylin and eosin (HE) and scored, and the liver was stained with oil red O to detect lipid accumulation. The integrity of the intestinal mucosal barrier was evaluated by measuring the excretion of diethylene triamine pentaacetic acid (DTPA). Western blotting was used to detect the expression of Tolllike receptor 2 (TLR2), Toll-like receptor 4 (TLR4), MyD88, nuclear factor-κB (NF-κB), ZO-1, Claudin-1, Occuludin-1, and PV-1 proteins in colon tissue; immunohistochemistry was used to analyze the expression of TLR4, MyD88, and NF-κB proteins in colon and liver tissue; and immunofluorescence was used to detect the expression of ZO-1, Claudin-1, and E-cadherin proteins. 16S rRNA gene sequencing was used to determine the changes in the types and structure of the intestinal microbiome. Results Compared with the model group, the levels of TG, TC, LDL-C, ALT, AST, and LPS in the serum of the XYS and PH groups were significantly lower (P < 0.05, 0.01, 0.001), and the levels of IL-1β, IL-6, and TNF-α in the liver were significantly lower (P < 0.01, 0.001). The level of HDL-C in the serum of the XYS high-dose group was significantly higher (P < 0.01). The scores of liver, colon, ileum, jejunum, and liver pathological grades were significantly lower in the XYS group, and the area stained with oil red O in the liver was significantly lower (P < 0.01, 0.001). The immunohistochemistry and Western blotting results showed that the expression of TLR2, TLR4, MyD88, and NF-κB proteins was significantly lower in the XYS and PH groups (P < 0.01, 0.001). The excretion rate of DTPA in 24 h urine of the rats in the XYS high-dose group and PH group was significantly lower (P < 0.05). In the high-dose group of XYS and the PH group, the expression of ZO-1, Claudin-1 and Occludin-1 proteins was significantly increased (P < 0.001), while the expression of PV-1 protein was significantly decreased (P < 0.001). Immunofluorescence showed that the expression of ZO-1, Claudin-1 and E-cadherin was significantly increased in the high-dose group of XYS and the PH group. Compared with the model group, XYS increased the richness and diversity of the intestinal microbiome, reduced the relative abundance of harmful bacteria such as Ruminococcus and increased the relative abundance of beneficial bacteria such as Bacteroides. Conclusion XYS has an ameliorative effect on MAFLD rats induced by high-fat diet, which can be achieved by alleviating liver inflammation, regulating liver function indicators. The mechanism may be related to repairing the intestinal mucosal barrier, regulating intestinal microbiota, and regulating the TLR4/MyD88/NF-κB pathway.

R285.5

常州市卫健委青年人才科技项目(QN202125);常州市卫生健康青苗人才培养工程资助项目(CZQM2022015)