人参皂苷F_(2)对胆汁淤积肝损伤小鼠抗炎、抗氧化、抗纤维化和抗凋亡的作用机制  

Study on mechanism of ginsenoside F_(2) on inflammation,oxidation,and fibrosis in mice with cholestatic liver injury

作　　者：徐华明[1] 杨柳 闫五玲 王培育[1] 郑思嘉 刘延鑫[1] 杨念[1] 张雪琳 聂闪闪 付银娜 邢伟鸽 王富利[2] XU Huaming;YANG Liu;YAN Wuling;WANG Peiyu;ZHENG Sijia;LIU Yanxin;YANG Nian;ZHANG Xuelin;NIE Shanshan;FU Yinna;XING Weige;WANG Fuli(Henan University of Chinese Medicine,Zhengzhou 450046,China;The Third Affiliated Hospital,Henan University of Chinese Medicine,Zhengzhou 450008,China;The First Affiliated Hospital,Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南中医药大学第三附属医院,河南郑州450008 [3]河南中医药大学第一附属医院,河南郑州450000

出　　处：《药物评价研究》2025年第1期38-50,共13页Drug Evaluation Research

基　　金：2024年度河南省中医药科学研究专项重点项目(2024ZY1030)。

摘　　要：目的探讨人参皂苷F_(2)(GF_(2))对α-萘异硫氰酸酯(ANIT)诱导的胆汁淤积肝损伤(CLI)小鼠的作用及机制。方法将70只雄性昆明小鼠随机分为7组(n=10):对照组、单给GF_(2)(100 mg·kg^(-1))组、模型组、熊去氧胆酸(UDCA,40 mg·kg^(-1))组和GF_(2)低、中、高剂量(25、50、100 mg·kg^(-1))组。小鼠连续ig给药7 d,于第5天ig给予ANIT(100 mg·kg^(-1))建立胆汁淤积模型。自动生化仪测量血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、总胆汁酸(TBA)、总胆红素(TBIL)、直接胆红素(DBIL)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSHPx)水平,肝组织匀浆上清MDA、SOD、GSH-Px和过氧化氢酶(CAT)水平;ELISA法检测肝组织匀浆上清液中炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-1β和脂多糖(LPS)水平;HE染色和Masson染色进行肝组织病理学分析;TUNEL染色观察肝细胞凋亡;免疫组化法观察免疫细胞标志物[中性粒细胞标志物CD11b和Ly6g、巨噬细胞标志物F4/80和T细胞标志物CD3]、肝纤维化标志物[α-平滑肌肌动蛋白(α-SMA)、I型胶原(Collagen I)]、Toll样受体4(TLR4)/髓分化因子88(MyD88)/核因子κB(NF-κB)相关蛋白、转化生长因子-β1(TGF-β1)/Smad相关蛋白、Bcl-2相关X蛋白(Bax)蛋白表达;Western blotting检测TLR4/Myd88/NF-κB、Nrf2/HO-1/NQO1和TGF-β1/Smad、B淋巴细胞瘤-2(Bcl-2)/Bax通路蛋白表达。结果与模型组比较,GF_(2)组AST、ALT、ALP、TBA、TBIL和DBIL水平显著降低(P<0.05、0.01、0.001),GF_(2)明显改善模型组肝细胞肿胀、空泡化、肝内炎症细胞浸润和坏死,胆管增生和扩张;GF_(2)显著降低ANIT诱导的炎症因子TNF-α、IL-6、IL-1β、LPS水平(P<0.05、0.01、0.001),明显降低CD11b、Ly6g、F4/80、CD3表达,显著降低TLR4、Myd88、NF-κB p65和pIκBα的蛋白表达(P<0.05、0.01);GF_(2)组MDA水平降低,SOD、CAT活性和GSH水平恢复,差异显著(P<0.05、0.01、0.00Objective To investigate the effect of ginsenoside F_(2)(GF_(2))onα-naphthalene isothiocyanate(ANIT)-induced cholestatic liver injury(CLI)in mice and its mechanism.Methods 70 male Kunming mice were randomly divided into seven groups(n=10):control group,single GF_(2)(100 mg∙kg^(-1))group,model group,UDCA(40 mg∙kg^(-1))group,and low,medium,and high dose GF_(2)(25,50,100 mg∙kg^(-1))groups.The mice were administered orally with the drugs for seven consecutive days,and ANIT(100 mg∙kg^(-1))was ig administered on the 5th day to establish a cholestasis model.The automatic biochemistry analyzer was used to measure serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bile acid(TBA),total bilirubin(TBIL),direct bilirubin(DBIL),malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)levels,and the levels of MDA,SOD,GSH-Px,and catalase in the liver tissue homogenate supernatant;the enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β,and LPS)in the liver tissue homogenate supernatant;hematoxylin and Masson staining were used for histopathological analysis of the liver tissue;TUNEL staining was used to observe hepatocyte apoptosis;immunohistochemistry was used to detect the expression of immune cell markers(neutrophil markers CD11b and Ly6g,macrophage markers F4/80 and T cell markers CD3),liver fibrosis markers(α-smooth muscle actin(α-SMA),type I collagen(Collagen I)),Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B(NF-κB)related protein,transforming growth factor-β1(TGF-β1)/SMAD-associated protein,Bcl-2 associated X protein(Bax)protein expression.Western blotting was used to detect the expression of TLR4/Myd88/NF-κB,Nrf2/HO-1/NQO1,TGF-β1/Smad,B-lymphoblastoma-2(Bcl-2)/Bax pathway proteins.Results Compared with the model group,the levels of AST,ALT,ALP,TBA,TBIL and DBIL were significantly lower in the GF_(

关 键 词：人参皂苷F_(2) 胆汁淤积 肝损伤 炎症 氧化应激 肝纤维化 凋亡 TLR4/Myd88/NF-κB通路 Nrf2/HO-1/NQO1通路 TGF-β1/Smad通路 

分 类 号：R285.5[医药卫生—中药学]