基于胆固醇代谢的复方甘草酸苷片配伍雷公藤多苷片减轻肝损伤机制  

Compound Glycyrrhizin Tablets Ameliorate Liver Injury Induced by Tripterygium Glycosides Tablet by Regulating Cholesterol Metabolism

作　　者：付小桐 曹春雨[1] 李春[1] 卢晨娜 刘婷[1] 杨依霏 FU Xiaotong;CAO Chunyu;LI Chun;LU Chenna;LIU Ting;YANG Yifei(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100029,China)

机构地区：[1]中国中医科学院中药研究所,北京100029

出　　处：《中国实验方剂学杂志》2025年第2期46-55,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：北京市自然科学基金项目(7232301);中国中医科学院创新工程项目(CI2021A04804);中央级公益性科研院所基本科研业务费专项(ZXKT22018,ZXKT21009,ZZ14-YQ-025)。

摘　　要：目的:基于胆固醇代谢,研究雷公藤多苷片(TG)的肝损伤作用机制,及复方甘草酸苷片(CG)配伍TG减轻TG所致胆固醇代谢异常的分子机制。方法:雄性SD大鼠按体质量随机分为正常组(纯净水),TG低、高剂量(TG-L、TG-H)组(TG 189.0、472.5 mg·kg^(-1)·d^(-1)),TG与复方甘草酸苷片配伍(TG-L+CG、TG-H+CG)组(给药剂量分别为TG 189.0 mg·kg^(-1)·d^(-1)+CG 20.25 mg·kg^(-1)·d^(-1)和TG 472.5 mg·kg^(-1)·d^(-1)+CG 20.25 mg·kg^(-1)·d^(-1)),每组6只,共5组。每日给药1次,连续3周。末次给药结束后,通过实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠肝组织内肝X受体-α(LXR-α)、低密度脂蛋白受体(LDLR)、三磷酸腺苷结合盒转运蛋白A1(ABCA1)、三磷酸腺苷结合盒转运蛋白G1(ABCG1)、3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)、胆固醇7α羟化酶(CYP7A1)、胆固醇12α羟化酶(CYP8B1)、甾醇27α羟化酶(CYP27A1)在信使核糖核酸(mRNA)水平的变化;利用蛋白免疫印迹法(Western blot)进一步验证关键蛋白表达水平的变化;通过酶联免疫吸附测定法(ELISA)检测胆固醇合成的调控酶羟甲基戊二酸单酰辅酶A还原酶(HMG-CoAR)的含量。以人肝癌细胞(HepG2)作为体外实验对象,观察TG对HepG2细胞的增殖抑制作用。设置TG提取物低、中、高剂量(TG-l、TG-m、TG-h)组(TG 135、45、15 mg·L^(-1)),非诺贝特(FB)组(10μmol·L^(-1))和CG提取物组,以及配伍组(TG-h+FB、TG-m+FB、TG-l+FB、TG-h+CG、TG-m+CG、TG-l+CG,给药浓度分别为TG 135 mg·L^(-1)+FB 10μmol·L^(-1)、TG 45 mg·L^(-1)+FB 10μmol·L^(-1)、TG 15 mg·L^(-1)+FB 10μmol·L^(-1)、TG 135 mg·L^(-1)+CG 60 mg·L^(-1)、TG 45 mg·L^(-1)+CG 60 mg·L^(-1)、TG 15 mg·L^(-1)+CG 60 mg·L^(-1)),通过Real-time PCR、Western blot检测HepG2细胞内LXR-α、ABCG1、LDLR、CYP7A1、CYP8B1、CYP27A1在mRNA水平及蛋白水平的表达变化,进一步验证CG配伍TG的减毒机制。结果:SD大鼠在体实验,与正常组比较,TG-H组大鼠肝组织CYPObjective:To investigate the mechanism of liver injury induced by tripterygium glycosides tablets(TG)and the molecular mechanism of compound glycyrrhizin tablets(CG)in alleviating the abnormalities of cholesterol metabolism caused by TG via cholesterol metabolism.Methods:According to the body weights,male Sprague-Dawley(SD)rats were randomly grouped as follows:control(pure water),low-dose TG(TG-L,189.0 mg·kg^(-1)·d^(-1)),high-dose TG(TG-H,472.5 mg·kg^(-1)·d^(-1)),TG-L+CG(189.0 mg·kg^(-1)·d^(-1)TG+20.25 mg·kg^(-1)·d^(-1)CG),and TG-H+CG(472.5 mg·kg^(-1)·d^(-1)TG+20.25 mg·kg^(-1)·d^(-1)CG),with 6 rats in each group.Rats were administrated with corresponding drugs once daily for 3 weeks.At the end of the last administration,the mRNA and protein levels of liver X receptor-alpha(LXR-α),low-density lipoprotein receptor(LDLR),adenosine triphosphate-binding cassette transporter A1(ABCA1),adenosine triphosphate-binding cassette transporter G1(ABCG1),3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGCR),cholesterol 7α-hydroxylase(CYP7A1),cholesterol 12α-hydroxylase(CYP8B1),and sterol 27-hydroxylase(CYP27A1)in the liver tissue were determined by Real-time PCR and Western blotting,respectively.The level of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMG-CoAR),a regulatory enzyme of cholesterol synthesis,was measured by enzyme-linked immunosorbent assay(ELISA).HepG2 cells were used to observe the effect of TG on the cell proliferation in vitro.Specifically,HepG2 cells were grouped as follows:Low-dose TG(TG-l,15 mg·L^(-1)),medium-dose TG(TG-m,45 mg·L^(-1)),high-dose TG(TG-h,135 mg·L^(-1)),fenofibrate(FB,10μmol·L^(-1)),CG extract,TG-h+FB(135 mg·L^(-1)TG+10μmol·L^(-1)FB),TG-m+FB(45 mg·L^(-1)TG+10μmol·L^(-1)FB),TG-l+FB(15 mg·L^(-1)TG+10μmol·L^(-1)FB),TG-h+CG(135 mg·L^(-1)TG+60μmol·L^(-1)CG),TG-m+CG(45 mg·L^(-1)TG+60μmol·L^(-1)CG),and TG-l+CG(15 mg·L^(-1)TG+60μmol·L^(-1)CG).The mRNA and protein levels of LXR-α,ABCG1,LDLR,CYP7A1,CYP8B1,and CYP27A1 in HepG2 cells were determined by Real-tim

关 键 词：雷公藤多苷片 复方甘草酸苷片 肝损伤 配伍减毒 胆固醇代谢 

分 类 号：R575[医药卫生—消化系统] R285[医药卫生—内科学] R256[医药卫生—临床医学]