杞黄葛复方对组织工程肝脏非酒精性脂肪性肝病模型的肝脏保护作用研究  

Study on the protective effect of Qihuangge compound on the non-alcoholic fatty liver disease model of tissue engineering liver

作　　者：黄龙 刘霜[1] 陈煜[1] 段钟平[1] HUANG Long;LIU Shuang;CHEN Yu;DUAN Zhongping(Beijing Key Laboratory of Liver Failure and Artificial Liver Treatment, Liver Diseases Center of Beijing You’an Hospital Affiliated to Capital Medical University, Beijing 100069, China)

机构地区：[1]首都医科大学附属北京佑安医院肝病中心/肝衰竭与人工肝治疗研究北京市重点实验室,北京100069

出　　处：《现代中西医结合杂志》2022年第5期591-595,704,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：大数据精准医疗高精尖创新中心项目(1212040205);北京市自然科学基金资助项目(7222094);首都卫生发展科研专项项目(2021-1G-2181)。

摘　　要：目的借助新型组织工程人工肝脏非酒精性脂肪性肝病模型,探讨杞黄葛复方对肝脏组织细胞氧化应激、脂肪酸代谢的影响。方法取大鼠肝脏,去细胞后加入人HepG2细胞制备组织工程肝脏。再细胞化完成后,实验分为3组:正常组用完全培养基培养;模型组用高脂培养基培养;杞黄葛复方组用高脂培养基培养,于第6天同时灌注100μg/mL的杞黄葛复方培养3 d。3组均于第8天培养结束收集细胞。采用CCK8法检测不同浓度(1 ng/mL、10 ng/mL、100 ng/mL、1μg/mL、10μg/mL、100μg/mL、1 mg/mL)的杞黄葛复方浓缩液干预后肝细胞存活率;按照试剂盒说明检测正常组、模型组和杞黄葛复方组HepG2细胞中三酰甘油(TG)、活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量;采用RT-qPCR检测模型组和杞黄葛复方组HepG2细胞中肝脏脂质合成途径相关酶[酸转位酶(CD36)、载脂蛋白B(ApoB)、脂肪酸合酶(FAS)、ATP柠檬酸裂解酶(ACLY)、脂肪酸去饱和酶2(FADS2)、乙酰辅酶A羧化酶(ACC)、脂肪酸结合蛋白1(FABP1)]和转录因子[过氧化物酶增殖物激活受体γ(PPARγ)]mRNA表达情况。结果杞黄葛复方各浓度组高脂培养的人HepG2细胞存活率与空白组比较差异均无统计学意义(P均>0.05)。模型组HepG2细胞中TG、ROS、MDA含量均明显高于正常组(P<0.05),SOD含量明显低于正常组(P<0.05);杞黄葛复方组HepG2细胞中TG、ROS、MDA含均明显低于模型组(P<0.05),SOD含量明显高于模型组(P<0.05)。杞黄葛复方组HepG2细胞中CD36、FAS、ACLY、FADS2、ACC、FABP1、PPARγmRNA相对表达量均明显低于模型组(P均<0.05),ApoB mRNA相对表达量与模型组比较差异无统计学意义(P>0.05)。结论杞黄葛复方可以通过下调肝脏脂质合成途径相关酶和转录因子的表达抑制游离脂肪酸的摄取,减少TG合成;可以通过减少ROS、MDA产生和提高SOD活性而增强细胞抗氧化能力,从而保护肝脏。Objective It is to explore the effect of Qihuangge compound on oxidative stress and fatty acid metabolism in liver tissue cells with the help of a non-alcoholic fatty liver disease model of new tissue-engineered artificial liver.Methods The rat livers were taken,decellularized and then added with human HepG2 cells to prepare tissue-engineered livers.After the recellularization was completed,the experiment was divided into 3 groups:the normal group was cultured with complete medium;the model group was cultured with high-fat medium;the Qihuangge compound group was cultured with high-fat medium,and 100μg/mL Qihuangge compound was simultaneously perfused on the 6th day.The cells were collected from the 3 groups at the end of the 8th day of culture.The survival rate of liver cells after intervention with different concentrations(1 ng/mL,10 ng/mL,100 ng/mL,1μg/mL,10μg/mL,100μg/mL,1 mg/mL)of Qihuangge compound concentrate were detected by CCK8 method;the contents of triacylglycerol(TG),reactive oxygen species(ROS),malondialdehyde(MDA)and superoxide dismutase(SOD)in HepG2 cells in the normal group,model group and Qihuangge compound group were detected according to the kit instructions;the mRNA expression of hepatic lipid synthesis pathway-related enzymes[acid translocase(CD36),apolipoprotein B(ApoB),fatty acid synthase(FAS),ATP citrate Lyase(ACLY),fatty acid desaturase 2(FADS2),acetyl-CoA carboxylase(ACC),fatty acid binding protein 1(FABP1)]and transcription factors[peroxidase proliferator-activated receptor gamma(PPARγ)]was detected by RT-qPCR.Results There was no significant difference in the survival rate of human HepG2 cells cultured with high fat in each concentration group of Qihuangge compound compared with the blank group(all P>0.05).The contents of TG,ROS and MDA in HepG2 cells of the model group were significantly higher than those of the normal group(all P<0.05),and the content of SOD was significantly lower than that of the normal group(P<0.05);the contents of TG,ROS and MDA in HepG2 cells of Qihuangge c

关 键 词：新型组织工程人工肝脏 非酒精性脂肪性肝病 杞黄葛复方 氧化应激 脂肪酸代谢 

分 类 号：R-332[医药卫生]