目的 基于核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）信号通路，研究黄芪赤风汤（HQCFT）对代谢相关脂肪性肝病（MAFLD）小鼠的作用机制。方法 将32只C57BL/6J小鼠采用高脂饲料与基础饲料混合饲喂，并在10 d内逐步减少基础饲料添加占比、增加高脂饲料占比，以达到适应性过渡喂养，后喂食高脂饲料持续12周，8只对照组小鼠接受正常饲料。将造模小鼠随机分为4组：模型组、阿托伐他汀钙片(Ato，阳性药，0.005 g·kg^-1)组和HQCFT低、高剂量(1.95、3.90 g·kg^-1)组，第13周开始ig给予各剂量HQCFT和Ato，持续28 d，而对照组和模型组给予等量的0.9%氯化钠溶液。观察小鼠一般状态；肝脏组织进行苏木素-伊红（HE）染色与油红O染色；试剂盒法检测血清总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、白细胞介素-1β（IL-1β）和IL-18含量；Western blotting分析测定肝脏p-核因子（NF）-κB/NF-κB、NLRP3、半胱氨酰天冬氨酸特异性蛋白酶-1（Caspase-1）以及凋亡相关斑点样蛋白（ASC）表达；通过Auto Dock软件对HQCFT中7种活性成分（芍药内酯苷、芍药苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮、芒柄花素、黄芪甲苷）和核心靶点（NLRP3、Caspase-1、ASC、NF-κB）进行分子对接。结果 与对照组相比，模型组出现明显活动量降低，毛发黏连，饮食在造模期较对照组增加，在造模成功后相较于对照组有一定减少，大便明显发臭，体质量上升；肝脏组织有明显脂肪空泡和炎症反应浸润；血清TC、LDL-C、HDL-C、AST、ALT、IL-1β、IL-18含量显著升高，肝脏组织中p-NF-κB/NF-κB、NLRP3、Caspase-1、ASC蛋白水平显著上调（P<0.05、0.01）；与模型组相比，HQCFT组小鼠状态好转，体质量降低，肝脏组织脂质沉积明显改善，炎症浸润减少，血清TC、LDL-C、HDL-C、AST、ALT、IL-1β、IL-18水平显著降低，肝组织pNF-κB/NF-κB、NLRP3、Caspase-1、ASC蛋白水平显著下调（P<0.05、0.01）。分子对接显示，HQCFT活性成分芍药内酯苷、芍药苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮、芒柄花素、黄芪甲苷与ASC、NF-κB、Caspase-1、NLRP3具有高亲和力。结论 HQCFT可明显改善高脂饮食诱导的MAFLD小鼠肝脏脂质沉积，与其抑制NLRP3炎症小体诱导的炎症反应密切相关。

Objective Based on the nucleotide-binding oligomerization domain（NOD）-like receptor 3（NLRP3） signaling pathway, the mechanism of Huangqi Chifeng Tang（HQCFT） on metabolic associated fatty liver disease（MAFLD） mice was studied. Methods Thirty-two C57 BL/6J mice were fed with a mixture of high-fat diet and basic diet. Over a period of 10 d, the proportion of the basic diet was gradually reduced and the proportion of the high-fat diet was increased to achieve an adaptive transition feeding. Then, the mice were fed with the high-fat diet for 12 weeks. Eight mice in the control group were given normal diet. The mice were randomly divided into four groups: the model group, the Ato(positive drug, 0.005 g·kg^-1) group, and the low and high doses of HQCFT(1.95 and 3.90 g·kg^-1) groups. From the 13 th week, each dose of HQCFT and Ato was ig given for 28 d, while the control group and the model group were given the same amount of 0.9% sodium chloride solution. The general condition of the mice was observed; liver tissues were stained with hematoxylin-eosin（HE） and oil red O; Serum total cholesterol（TC）, triglyceride（TG）, low-density lipoprotein cholesterol（LDL-C）, high-density lipoprotein cholesterol（HDL-C）, alanine aminotransferase（ALT）, aspartate aminotransferase（AST）, interleukin-1β（IL-1β） and IL-18 contents were detected by kits; Western blotting was used to analyze the expression of p-NF-κB/NF-κB, NLRP3, cysteinyl aspartate-specific protease-1（Caspase-1）, and ASC proteins in the liver; the seven active components in HQCFT（paeonol glycoside, paeonol, anthocyanidin glucoside of sophorae, sophorin, anthocyanidin, sophoroside, astragaloside） and the core targets [NLRP3, Caspase-1, apoptosis-related speck-like protein（ASC）, NF-κB] were subjected to molecular docking using Auto Dock software. Results Compared with the control group, the model group showed significant reduction in activity level, hair adhesion, increased food intake during the modeling period, a certain decrease compared to the control group after successful modeling, significantly more foul-smelling feces, and increased body weight; Liver tissue showed obvious fat vacuoles and inflammatory infiltration; The contents of serum TC, LDL-C, HDL-C, AST, ALT, IL-1β, and IL-18 significantly increased, and the protein levels of p-NF-κB/NF-κB, NLRP3, Caspase-1, and ASC in liver tissue significantly upregulated（P < 0.05, 0.01） compared with the model group; compared with the model group, mice in the HQCFT group improved their condition, had lower body weight, significantly improved lipid deposition in liver tissue, reduced inflammatory infiltration, and significantly lower serum TC, LDL-C, HDL-C, AST, ALT, IL-1β, and IL-18 levels, and significantly downregulated the protein levels of p-NF-κB/NF-κB, NLRP3, Caspase-1, and ASC in liver tissue（P < 0.05, 0.01）. Molecular docking showed that the active components of HQCFT, such as peucedane glycoside, peucedanin, anthocyanidin glucoside, penicilin glycoside, anthocyanidin, penicilin, and astragaloside IV, had high affinity with ASC, NF-κB, Caspase-1, and NLRP3. Conclusion HQCFT can significantly improve lipid deposition in the liver of mice with MAFLD induced by high-fat diet, and is closely related to its inhibition of the inflammatory response induced by NLRP3 inflammasome.

R285.5

黑龙江省自然科学基金项目(PL2024H239)