目的 研究逍遙散對(duì)高脂飲食誘導(dǎo)的大鼠代謝相關(guān)脂肪性肝?。∕AFLD）的保護(hù)作用及機(jī)制。方法 SD大鼠隨機(jī)分為5組：對(duì)照組、模型組、鹽酸吡格列酮（PH，陽(yáng)性藥，5 mg·kg^-1）和逍遙散低、高劑量（19、38 g·kg^-1）組。對(duì)照組喂食基礎(chǔ)日糧，而其余組喂食高脂肪和高果糖日糧12周，從第13周開(kāi)始ig給藥，每天1次持續(xù)4周。試劑盒法檢測(cè)血清三酰甘油（TG）、總膽固醇（TC）、高密度脂蛋白膽固醇（HDL-C）、低密度脂蛋白膽固醇（LDL-C）、丙氨酸氨基轉(zhuǎn)移酶（ALT）、天冬氨酸氨基轉(zhuǎn)移酶（AST）、內(nèi)毒素（LPS）水平，肝組織白細(xì)胞介素-6（IL-6）、IL-1β、腫瘤壞死因子-α（TNF-α）水平；HE染色觀察肝臟、結(jié)腸、空腸和回腸組織病理?yè)p傷并評(píng)分，油紅O染色觀察肝臟脂質(zhì)累積；檢測(cè)24 h尿液99mTc-噴替酸（DTPA）排泄評(píng)價(jià)腸黏膜屏障完整性；Western blotting法檢測(cè)結(jié)腸組織toll樣受體2（TLR2）、toll樣受體4（TLR4）、髓樣分化因子88（MyD88）、核因子-κB（NF-κB）、閉鎖小帶蛋白-1（ZO-1）、封閉蛋白1（Claudin-1）、緊密連接蛋白-1（Occuludin-1）、質(zhì)膜囊泡相關(guān)蛋白1（PV-1）蛋白表達(dá)；免疫組化分析結(jié)腸和肝組織TLR4、MyD88、NF-κB蛋白表達(dá)；免疫熒光檢測(cè)ZO-1、Claudin-1、E-鈣黏蛋白（E-cadherin）蛋白表達(dá)。通過(guò)16S rRNA基因測(cè)序確定腸道菌群種類變化和結(jié)構(gòu)。結(jié)果 與模型組相比，逍遙散和PH組血清中TG、TC、LDL-C、ALT、AST和LPS水平顯著降低（P＜0.05、0.01、0.001），肝臟IL-1β、IL-6、TNF-α水平顯著降低（P＜0.01、0.001），逍遙散高劑量組血清HDL-C水平顯著升高（P＜0.01）；逍遙散組肝臟、結(jié)腸、空腸、回腸和肝臟病理評(píng)分顯著降低，肝臟油紅O染色區(qū)域占比顯著降低（P＜0.01、0.001）；免疫組化與Western blotting結(jié)果顯示，逍遙散和PH組TLR2、TLR4、MyD88、NF-κB蛋白表達(dá)顯著降低（P＜0.01、0.001）；逍遙散高劑量和PH組大鼠的24 h尿液DTPA排泄顯著降低（P＜0.05）；逍遙散高劑量組和PH組ZO-1、Claudin-1和Occludin-1蛋白表達(dá)顯著升高（P＜0.001），PV-1蛋白表達(dá)顯著降低（P＜0.001），免疫熒光顯示逍遙散高劑量組和PH組的ZO-1、Claudin-1和E-cadherin的表達(dá)明顯升高。與模型組比較，逍遙散增加腸道菌群豐富度和多樣性，減少瘤胃球菌等有害菌、增加擬桿菌等有益菌的相對(duì)豐度。結(jié)論 逍遙散對(duì)高脂飲食誘導(dǎo)的MAFLD大鼠具有改善作用，可通過(guò)減輕肝臟炎癥、調(diào)節(jié)肝功能指標(biāo)，機(jī)制可能與修復(fù)腸道黏膜屏障、調(diào)節(jié)腸道菌群、調(diào)節(jié)TLR4/MyD88/NF-κB通路等相關(guān)。

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

常州市衛(wèi)健委青年人才科技項(xiàng)目(QN202125);常州市衛(wèi)生健康青苗人才培養(yǎng)工程資助項(xiàng)目(CZQM2022015)