藏药波棱瓜子总木脂素对ANIT诱导的小鼠肝损伤的保护作用及机制研究  

Study on Protective Effect and Mechanism of Total Lignans From Tibetan Medicine Herpetospermun caudigerum Wall Against α-Naphthylisothiocyanate-Induced Liver Injury in Mice

作　　者：李娜芝 师艺玮 郭敏侠 朱继孝 梁健 冯育林 LI Nazhi;SHI Yiwei;GUO Minxia;ZHU Jixiao;LIANG Jian;FENG Yulin(Research Center for Traditional Chinese Medicine Resource and Ethnic Minority Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004 Jiangxi,China)

机构地区：[1]江西中医药大学,中药资源与民族药研究中心,江西南昌330004

出　　处：《中药新药与临床药理》2023年第1期49-56,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家重点研发计划项目(2019YFC1712302)。

摘　　要：目的 探讨波棱瓜子总木脂素(TLHPS)对α-萘异硫氰酸酯(ANIT)诱导的小鼠肝内胆汁淤积(intrahepatic cholestasis,IC)型肝损伤的保护作用及其作用机制。方法 将70只小鼠随机分为正常组、模型组、联苯双酯组、TLHPS各剂量组,各组连续给药7 d,在第5天给药2 h后,除正常组灌胃橄榄油溶液外,其余各组小鼠给予ANIT溶液造模。之后按照原剂量继续给药,造模48 h后,对各组小鼠进行摘眼球采血,并摘取肝脏。检测各组小鼠血清中肝功能指标谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、直接胆红素(DBIL)、总胆红素(TBIL)、总胆汁酸(TBA);炎症因子肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)及肝脏抗氧化应激因子谷胱甘肽(GSH-PX)、超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT);采用HE染色检测肝脏病理变化;Western Blot法检测小鼠肝组织中NLRP3、TLR-2、TLR-4、NF-κB、JNK、p38、MyD88、IL-1β蛋白表达水平。结果 与正常组比较,模型组肝脏受损情况严重,ALT、AST、ALP酶活力明显升高(P<0.01),DBIL、TBIL、TBA、MDA、TNF-α、MCP-1含量明显升高(P<0.01),CAT、GSH-PX、SOD酶活性明显降低(P<0.01),NLRP3、TLR-2、TLR-4、NF-κB、JNK、p38、MyD88、IL-1β蛋白表达水平明显升高(P<0.01)。与模型组比较,联苯双酯组及TLHPS各给药组ALT、AST、ALP酶活力明显降低(P<0.01),DBIL、TBIL、TBA、MDA、TNF-α、MCP-1含量明显降低(P<0.01),CAT、GSH-PX、SOD酶活性明显升高(P<0.05,P<0.01),NLRP3、TLR-2、TLR-4、NF-κB、JNK、p38、MyD88、IL-1β蛋白表达水平明显降低(P<0.05,P<0.01)。与联苯双酯组比较,TLHPS(0.4 g·kg^(-1))组ALT、AST、ALP、TBA、TBIL、DBIL含量明显升高(P<0.05,P<0.01),TLHPS各给药组SOD含量明显降低(P<0.01),TLHPS(0.05 g·kg^(-1))给药组JNK、p38、IL-1β蛋白表达水平明显上调(P<0.05,P<0.01),TLHPS(0.4、0.2、0.1 g·kg^(-1))给药组NF-κB蛋白表达明显上调(P<0.05,P<0.01),其余各组差异�Objective To investigate the protective effect of total lignans from Herpetospermun caudigerm Wal(TLHPS) on α-naphthylisothiocyanate(ANIT)-induced intrahepatic cholestatic(IC) liver injury in mice and its mechanism of action.Methods Seventy mice were randomly and evenly divided into normal group,model group,Bifendate Pills group and TLHPS group.All groups were administered continuously for 7 days.After 2-hour of administration on the 5th day,mice in the other groups were modeled by gavage with ANIT solution except for mice in the normal group,which were gavaged with olive oil solution.Then the mice were continued to be administered at the original dose.After 48-hour of modeling,blood was collected from the eyes and the liver was removed in each group.Serum liver function indexes of mice alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),direct bilirubin(DBIL),total bilirubin(TBIL)and total bile acid(TBA),inflammatory factors tumor necrosis factor-α(TNF-α),monocyte chemoattractant protein-1(MCP-1),and liver antioxidant stress factors glutathione(GSH-PX),superoxide dismutase(SOD),malondialdehyde(MDA)and catalase(CAT)in each group were detected.The pathological changes of liver were detected by HE staining.The expression levels of NLRP3,TLR-2,TLR-4,NF-κB,JNK,p38,MyD88 and IL-1β in mice liver tissue were detected by Western Blot.Results Compared with the normal group,severe liver damage was observed in the model group,and the increase in ALT,AST and ALP enzyme activities was found(P<0.01).The content of DBIL,TBIL,TBA,MDA,TNF-α and MCP-1 were significantly increased(P<0.01),while CAT,GSH-PX and SOD enzyme activities were significantly decreased(P<0.01).The protein expression levels of NLRP3,TLR-2,TLR-4,NF-κB,JNK,p38,MyD88 and IL-1β were significantly increased(P<0.01).Compared with the model group,ALT,AST and ALP enzyme activities were significantly reduced in Bifendate Pills group and each TLHPS administration group(P<0.01),the content of DBIL,TBIL,TBA,MDA,TNF-α and MCP-1

关 键 词：波棱瓜子 总木脂素 α-萘异硫氰酸酯(ANIT) 胆汁淤积 肝损伤 炎症 作用机制 氧化应激 小鼠 

分 类 号：R285.5[医药卫生—中药学]