健肝消脂方通过GPx4信号通路对非酒精性脂肪肝小鼠铁死亡的调控机制研究  

Study on Regulatory Mechanism of Jiangan Xiaozhi Prescription on Ferroptosis in Non-Alcoholic Fatty Liver Disease Mice Through GPx4 Signaling Pathway

作　　者：史玉龙 廖加抱 张能华[3] SHI Yulong;LIAO Jiabao;ZHANG Nenghua(Intensive Care Unit,Jiaxing Hospital of Traditional Chinese Medicine,Jiaxing Zhejiang 314001,China;Graduate School of Yunnan University of Chinese Medicine,Kunming Yunnan 650500,China;Ministry of Science and Education,Jiaxing Hospital of Traditional Chinese Medicine,Jiaxing Zhejiang 314001,China)

机构地区：[1]嘉兴市中医医院重症医学科,浙江嘉兴314001 [2]云南中医药大学研究生院,云南昆明650500 [3]嘉兴市中医医院科教部,浙江嘉兴314001

出　　处：《新中医》2025年第5期209-214,共6页New Chinese Medicine

基　　金：嘉兴市科技计划项目(2023AY11035)。

摘　　要：目的:观察健肝消脂方对非酒精性脂肪肝(NAFLD)模型小鼠的治疗效果及通过谷胱甘肽过氧化物酶4(GPx4)信号通路对铁死亡的影响。方法:60只C57BL/6J小鼠随机选取50只给予高脂饮食诱导建立NAFLD模型,另外10只作为正常组给予普通饮食。将造模后的小鼠分为模型组、抑制剂组及中药低、中、高剂量组,每组10只。以灌胃的方式对药物处理组给予不同浓度的药物,正常组和模型组分别给予等体积的0.9%氯化钠溶液。检测各组小鼠体质量、肝指数、血脂指标,苏木素-伊红(HE)染色肝石蜡切片评估健肝消脂方对NAFLD模型小鼠的治疗作用;通过对丙二醛(MDA)、4-羟基壬烯醛(4-HNE)、活性氧(ROS)水平的检测来评估健肝消脂方对NAFLD模型小鼠脂质过氧化水平的影响;通过检测总铁离子(Fe)水平以及转铁蛋白(Transferrin)、前列腺六段跨膜上皮抗原1(Steap1)、铁蛋白重链1(FTH1)、GPx4蛋白水平来评估健肝消脂方对NAFLD模型小鼠铁死亡及GPx4信号通路的影响。结果:正常组小鼠肝板排列整齐,结构正常;模型组小鼠肝板排列紊乱且有大量炎细胞浸润,提示肝脏具有严重脂肪变性;抑制剂组和中药高剂量组的病理缓解最为明显。与正常组比较,模型组小鼠体质量、肝组织中Fe含量降低(P<0.05),肝指数、谷丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、甘油三酯(TG)、总胆固醇(TC)、MDA、ROS以及4-HNE水平升高(P<0.05)。与模型组比较,抑制剂组、中药中剂量组、中药高剂量组小鼠体质量、肝组织中Fe含量升高(P<0.05),肝指数、ALT、AST、TG、TC、MDA、ROS以及4-HNE水平降低(P<0.05)。qPCR结果显示,与正常组比较,模型组小鼠Steap1、Transferrin表达升高,FTH1、GPx4表达降低(P<0.05)。与模型组比较,抑制剂组、中药中剂量组、中药高剂量组小鼠Steap1、Transferrin表达降低,FTH1、GPx4表达升高(P<0.05)。Western blotting结果显示,与正常组比较Objective:To observe the therapeutic effect of Jiangan Xiaozhi Prescription on non-alcoholic fatty liver disease(NAFLD)model mice and its impact on ferroptosis through the glutathione peroxidase 4(GPx4)signaling pathway.Methods:A total of 60 C57BL/6J mice were randomly selected,with 50 mice being induced to establish a NAFLD model with a high-fat diet,and the other 10 mice being treated as the normal group with a regular diet.After modeling,the mice were divided into the model group,the inhibitor group,and the Chinese medicine groups with low-,medium-,and high-doses,with 10 mice in each group.The Chinese medicine groups and the inhibitor group were given different concentrations of drugs by gavage,and the normal group and the model group were given equal volumes of 0.9%sodium chloride solution.Detected the body mass,liver index,and blood lipid indicators of each group of mice,and evaluated the therapeutic effect of Jiangan Xiaozhi Prescription on NAFLD model mice by staining liver paraffin sections with hematoxylin eosin(HE).Evaluated the effect of Jiangan Xiaozhi Prescription on lipid peroxidation levels in NAFLD model mice by detecting the levels of malondialdehyde(MDA),4-hydroxynonenal(4-HNE),and reactive oxygen species(ROS).The effects of Jiangan Xiaozhi Prescription on ferroptosis and GPx4 signaling pathway in NAFLD model mice were evaluated by detecting the levels of total Ferric ions(Fe),transferrin,six transmembrane epithelial antigen of the prostate 1(STEAP1),ferritin heavy chain 1(FTH1),and GPx4 protein.Results:The liver plates of the normal group mice were arranged neatly and structurally normal;the liver plate arrangement of the model group mice was disordered and there was a large amount of inflammatory cell infiltration,indicating severe hepatic steatosis;the pathological relief was most significant in the inhibitor group and the high-dose Chinese medicine group.Compared with the normal group,the body mass and Fe content in the liver tissue in the model group mice were decreased(P<0.05),while the li

关 键 词：非酒精性脂肪肝 健肝消脂方 铁死亡 脂质过氧化 谷胱甘肽过氧化物酶4 

分 类 号：R285.5[医药卫生—中药学]