白花蛇舌草对ANIT诱导的胆汁淤积大鼠保肝作用研究  

Study on Hepatoprotective Effect of Hedyotidis diffusae Herba on ANIT-induced Cholestasis in Rats

作　　者：钟艳花 李颖仪[2] 林重[1] 钟映芹 唐东晖 郭洁文 刘昌辉[2] ZHONG Yanhua;LI Yingyi;LIN Zhong;ZHONG Yingqin;TANG Donghui;GUO Jiewen;LIU Changhui(Guangzhou Liwan Hospital of Traditional Chinese Medicine,Guangzhou 510140 Guangdong,China;Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Guangzhou Hospital of Traditional Chinese Medicine,Guangzhou 510130 Guangdong,China)

机构地区：[1]广州市荔湾区中医医院,广东广州510140 [2]广州中医药大学,广东广州510405 [3]广州市中医院,广东广州510130

出　　处：《中药新药与临床药理》2021年第7期952-958,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81773936;81102883);广州市科技计划民生科技重点项目(202002020032)。

摘　　要：目的研究白花蛇舌草对α-萘异硫氰酸酯(ANIT)诱导的胆汁淤积肝病大鼠的保肝作用及机制。方法将48只SD大鼠随机分为正常对照组,模型组,白花蛇舌草高、中、低剂量组(1、0.5、0.25 g·kg^(-1)),熊去氧胆酸组(100 mg·kg^(-1)),每组8只,均连续灌胃给予相应药物5 d。其中第3天除正常对照组外,其他各组采用灌胃给予ANIT(65 mg·kg^(-1))制备ANIT诱导胆汁淤积肝病大鼠模型。检测各组大鼠血清肝功指标谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP),血清黄疸指数总胆汁酸(TBA)、总胆红素(TBIL)及肝脏抗氧化应激因子谷胱甘肽(GSH)和超氧化物歧化酶(SOD)水平;测定各组大鼠胆汁流量以及胆汁中TBIL和TBA的含量;采用HE染色检测肝脏病理变化;Real-time PCR法检测肝脏组织中胆汁酸合成、转运、排泄和重吸收等相关基因的表达水平。结果与正常对照组大鼠相比,模型组大鼠血清的ALT、AST、ALP、TBA、TBIL水平显著增加,而胆汁流量和胆汁中TBA和TBIL的排泄则显著降低,差异均具有统计学意义(P<0.01);胆汁酸合成与转运的主要核受体(FXR、SHP)、胆汁酸顶腔侧膜转运体(BSEP、MRP2、MDR2)、胆汁酸合成酶(CYP7A1、CYP8B1、CYP27A1)和胆汁酸重吸收转运体(NTCP、OATP1A1、OATP1A2)的mRNA表达水平显著降低,而胆汁酸基底侧膜转运体(MRP3、MRP4、OSTα、OSTβ)的mRNA表达水平则显著升高,差异均具有统计学意义(P<0.05,P<0.01)。与模型组相比,白花蛇舌草组大鼠血清的ALT、AST、ALP、TBA、TBIL水平显著降低(P<0.01),而大鼠胆汁流量和胆汁中TBA和TBIL的排泄量则显著升高(P<0.01);肝脏组织病理损伤显著改善;胆汁酸核受体(FXR、SHP)、胆汁酸顶腔侧膜转运体(BSEP、MRP2、MDR2)和胆汁酸基底侧膜转运体(MRP3、MRP4、OSTα、OSTβ)的mRNA表达水平显著升高(P<0.05,P<0.01),而胆汁酸合成酶(CYP7A1)和胆汁酸重吸收转运体(NTCP、OATP1A1)的mRNA表达水平显著降低(P<0.05Objective To study the hepatoprotective effect and mechanism of Hedyotidis diffusae herba(HHD)onα-naphthyl isothiocyanate(ANIT)induced cholestatic liver injury in rats.Methods The cholestatic liver injury rat model was induced by intragastric administration of ANIT(65 mg·kg^(-1)).Rats were randomly divided into control group,ANIT group,HHD high,medium and low dose groups(1,0.5,0.25 g·kg^(-1))and UDCA group(100 mg·kg^(-1)).The serum liver function indexes(ALT,AST and ALP),serum jaundice indexes(TBA,TBIL)and liver antioxidant stress factors(GSH,SOD)were detected.The bile flow and the contents of bilirubin(TBIL)and bile acid(TBA)in bile were determined.The pathological changes of liver were detected by HE staining.The mRNA expression of genes involved in bile acid synthesis,transport,excretion and reabsorption were detected by real-time PCR.Result Compared with the control group,the serum levels of ALT,AST,ALP,TBA and TBIL of rats in ANIT group were significantly increased,while the bile flow and the excretion of TBA and TBIL in bile were significantly decreased(P<0.01);the mRNA expression levels of the main nuclear receptors associated with bile acid synthesis and transport(FXR,SHP),bile acid apical membrane transporters(BSEP,MRP2,MDR2),bile acid synthase(CYP7A1,CYP8B1,CYP27A1)and bile acid reabsorption transporters(NTCP,OATP 1A1,OATP1A2)were significantly decreased,while the mRNA expression levels of bile acid basolateral membrane transporters(MRP3,MRP4,OSTα,OSTβ)were significantly increased(P<0.05,P<0.01).Compared with ANIT group,the levels of ALT,AST,ALP,TBA and TBIL in serum of rats in Hedyotidis diffusae herba treated group were significantly decreased(P<0.01),while the bile flow and the excretion of TBA and TBIL in bile were significantly increased(P<0.01).The pathological damage of liver tissue was significantly improved;the mRNA expression levels of bile acid nuclear receptor(FXR,SHP),bile acid apical membrane transporter(BSEP,MRP2,MDR2)and bile acid basolateral membrane transporters(MRP3,MRP4,OSTα,O

关 键 词：白花蛇舌草 胆汁淤积 Α-萘异硫氰酸酯 胆汁酸 大鼠 

分 类 号：R285.5[医药卫生—中药学]