基于Nrf2信号通路探究参苓白术散调节铁死亡对酒精性肝损伤大鼠的影响  

Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway

作　　者：周相宇 周素芳 李玉茹 蔡世揿 廖佳佳 叶佐玉 ZHOU Xiangyu;ZHOU Sufang;LI Yuru;CAI Shiqin;LIAO Jiajia;YE Zuoyu(The First Clinical Medical College of Guizhou University of Traditional Chinese Medicine(TCM),Guiyang 550002,China;First Afiliated Hospital of Guizhou University of TCM,Guiyang 550001,China)

机构地区：[1]贵州中医药大学第一临床医学院,贵阳550002 [2]贵州中医药大学第一附属医院,贵阳550001

出　　处：《中国实验方剂学杂志》2023年第5期104-113,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81460683);贵州省科技项目计划项目(黔科合支撑[2021]一般015)。

摘　　要：目的:观察参苓白术散通过调节核因子E2相关因子2(Nrf2)信号通路的干预调节铁死亡对酒精性肝损伤大鼠的影响。方法:将40只SD大鼠随机分为模型组、多烯磷脂酰胆碱组、参苓白术散高、中、低剂量组,每组8只,另取8只SD大鼠作为正常组。模型组、多烯磷脂酰胆碱组、参苓白术散高、中、低剂量组予以10 mL·kg^(-1)灌胃造模,正常组予以等体积蒸馏水灌胃;每日灌酒4 h后,予以多烯磷脂酰胆碱组143.64 mg·kg^(-1)药物灌胃、参苓白术散高、中、低剂量组分别予以15、7.5、3.75 mg·kg^(-1)的药物,正常组与模型组予以等体积蒸馏水灌胃,灌胃连续6周。全自动生化仪检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、谷氨酰转肽酶(GGT)、总胆固醇(TC)、甘油三酯(TG)水平,酶联免疫吸附测定法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平,生化检测法检测脂多糖(LPS)、丙二醇(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、铁离子(Fe^(+))水平,苏木素-伊红(HE)和油红O染色观察肝脏病理变化,实时荧光定量聚合酶链式反应(Real-time PCR)检测Nrf2、血红素加氧酶-1(HO-1)、谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链1(FTH1)、核转录因子-κB(NF-κB) mRNA表达,蛋白免疫印迹法(Western blot)检测Nrf2、HO-1、GPX4、FTH1、p65、磷酸化(p)-p65蛋白表达。结果:与正常组比较,模型组肝功能(ALT、AST、GGT)、血脂(TC、TG)水平明显升高(P<0.05),肝脏出现明显脂肪变性,有大量脂肪沉积,氧化应激水平及炎症因子明显升高(P<0.05),Fe^(+)水平明显增高(P<0.05),Nrf2、HO-1、GPX4、FTH1蛋白表达明显下调(P<0.05),p65、p-p65蛋白表达明显上调(P<0.05),Nrf2、HO-1、GPX4、FTH1 mRNA表达明显下调(P<0.05),NF-κB mRNA表达明显上调(P<0.05);与模型组比较,参苓白术散高、中剂量组肝功能(ALT、AST、GGT)、血脂(TC、TG)水平明显下调(P<0.05),肝脏脂肪变性明显改Objective:To observe the effect of Shenling Baizhusan on the intervention of the nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathway by regulating ferroptosis in rats with alcoholic liver injury. Method:Forty SD rats were randomly divided into model group, polyene phosphatidylcholine group, and high, medium, and low-dose Shenling Baizhusan groups, with 8 rats in each group. Another 8 SD rats were taken as blank group. The model group, polyene phosphatidylcholine group, high, medium, and low-dose Shenling Baizhusan groups were given 10 mL·kg^(-1)liquor by gavage for modeling, and the blank group was given equal volume of distilled water by gavage. After 4 h of daily alcoholic administration, 143.64 mg·kg^(-1)of polyene phosphatidylcholine group was given to the polyene phosphatidylcholine group, 15, 7.5, 3.75 mg·kg^(-1)of Shenling Baizhusan were given to Shenling Baizhusan high, medium, and low-dose groups, respectively, and the blank group and the model group were given equal volume of distilled water. The gavage lasted for 6weeks. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), glutamyl transpeptidase(GGT), total cholesterol(TC), and triglyceride(TG) were detected by automatic biochemical analyzer. The levels of tumor necrosis factor-α(TNF-α) and interleukin-β(IL-β) were detected by the enzymelinked immunosorbent assay(ELISA). The levels of lipopolysaccharide(LPS), malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and Fe^(+)were detected by biochemical assay. The pathological changes in the liver were observed by hematoxylin-eosin(HE) staining and oil red O staining. The mRNA expression levels of Nrf2, heme oxygenase-1(HO-1), glutathione peroxidase 4(GPX4), ferritin heavy polypeptide 1(FTH1), and nuclear factor-κB(NF-κB) were detected by Real-time polymerase chain reaction(Real-time PCR). The protein expression levels of Nrf2, HO-1, GPX4, FTH1, p65, and phosphorylation(p)-p65 were detected by Western blot. Result:As compared with the blank g

关 键 词：参苓白术散 酒精性肝损伤 核因子E2相关因子2(Nrf2) 氧化应激 铁死亡 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33