基于Nrf2/SLC7A11/GPX4通路探讨葛根芩连汤通过抑制铁死亡改善非酒精性脂肪性肝病作用机制  

Mechanism of action of Gegen QinLian Decoction in improving non-alcoholic fatty liver disease by inhibiting ferroptosis based on the Nrf2/SCLC7A11/GPX4 pathway

作　　者：骆倩 罗涛[2] 宋针珍 梁芳 黎俊森 彭淑红 曹岚 赵海平 章常华[3] LUO Qian;LUO Tao;SONG Zhenzhen;LIANG Fang;LI Junsen;PENG Shuhong;CAO Lan;ZHAO Haiping;ZHANG Changhua(Jiangxi Provincial Engineering Research Center of Development and Evaluation of TCM Classic Prescriptions,Nanchang 330004,China;the First Affiliated Hospital of Nanchang University,Nanchang 330006;School of Pharmacy,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004;School of Physical Education,Yuzhang Normal University,Nanchang 330103;Jiangxi Key Laboratory of TCM Etiology and Biology,Nanchang 330004;Research Center of Traditional Chinese Medicine Resources and Ethnomedicine,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004)

机构地区：[1]江西省中医经典名方(验方)开发与评价技术工程研究中心,南昌330004 [2]南昌大学第一附属医院,南昌330006 [3]江西中医药大学药学院,南昌330004 [4]豫章师范学院体育学院,南昌330103 [5]江西省中医病因生物学重点实验室,南昌330004 [6]江西中医药大学中药资源与民族药研究中心,南昌330004

出　　处：《中国比较医学杂志》2025年第2期72-84,共13页Chinese Journal of Comparative Medicine

基　　金：赣鄱俊才-江西省主要学科学术和技术带头人培养项目(领军人才-学术类)(20232BCJ22022);国家自然科学基金(82060741);江西省教育厅科技项目(GJJ213111)。

摘　　要：目的基于核转录因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)/谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)信号通路,通过体内外实验研究葛根芩连汤治疗非酒精性脂肪性肝病(NAFLD)的作用机制。方法以高脂饲料长期喂养大鼠24周完成NAFLD的诱导建模,成模大鼠随机分为正常组、模型组、葛根芩连汤高、中、低剂量组和二甲双胍组。自第25周起,按照分组情况灌胃相应药物两周直至取材。生化试剂盒检测各组大鼠肝组织氧化应激水平:丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)。Fe^(2+)试剂盒检测各组大鼠肝组织Fe^(2+)含量。实时荧光定量逆转录PCR(qRT-PCR)法检测各组大鼠肝组织Nrf2、血红素加氧酶1(heme oxygenase-1,HO-1)、SLC7A11、谷胱甘肽合成酶(glutathione synthetase,GSS)、GPX4和酰基辅酶A合成化酶4(acyl coenzyme A synthase,ACSL4)mRNA的表达水平。采用1 mmol/L游离脂肪酸(FFA)诱导肝癌HepG2细胞产生脂质堆积模拟NAFLD构建体外模型,加入不同浓度葛根芩连汤和二甲双胍含药血清进行治疗。油红O染色检测各组细胞脂质堆积情况。试剂盒测定不同组HepG2细胞MDA和GSH含量。细胞专用Fe^(2+)试剂盒检测各组细胞Fe^(2+)含量。qRT-PCR法检测各组细胞中Nrf2、HO-1、SLC7A11、GSS、GPX4和ACSL4 mRNA的表现水平。结果动物实验中,模型组大鼠肝组织MDA和Fe^(2+)含量较正常组显著升高,GSH含量显著降低(P<0.01);葛根芩连汤高、中剂量组和二甲双胍组可大幅度降低MDA和Fe^(2+)含量,升高GSH含量,与模型组比较(P<0.01,P<0.05)。与模型组相比,葛根芩连汤高、中剂量组和二甲双胍组均能增加Nrf2、HO-1、GSS、GPX4 mRNA的表达水平,降低ACSL4 mRNA的水平(P<0.01,P<0.05)。在细胞实验中,HepG2细胞模型组红色脂滴较正常组明显增多,葛根芩连汤和二甲双胍的使用导致脂滴明显减少。试�Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.MethodsRats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe^(2+))in rat liver tissues was detected using a Fe^(2+)kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid accumulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.ResultsIn the animal experiments,MDA and Fe^(2+)liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P<0.01).GGQLT-H,GGQLT

关 键 词：葛根芩连汤 非酒精性脂肪性肝病 铁死亡 氧化应激 Nrf2/SLC7A11/GPX4通路 

分 类 号：R-33[医药卫生]