中药新药与临床药理2025，Vol.36Issue(4)：506-514,9.DOI:10.19378/j.issn.1003-9783.2025.04.003
基于CYP酶/Caspase-3途径探讨何首乌及其主要成分致药物性肝损伤的作用机制
Mechanistic Study on the Mechanism of Drug-Induced Liver Injury Induced by Polygoni Multiflori Radix and Its Main Components Based on the CYP Enzyme/Caspase-3 Pathway
摘要
Abstract
Objective To explore the mechanism of drug-induced liver injury(DILI)induced by Polygoni Multiflori Radix and its main components based on CYP enzyme/Caspase-3 pathway.Methods(1)In vitro experiment:HepG2 cells were divided into blank group,emodin low-,medium-and high-dose groups(50,100,150 µmol·L-1),rhein low-,medium-and high-dose groups(100,150,200 µmol·L-1),gallic acid low-,medium-and high-dose groups(100,150,200 µmol·L-1),and stilbene glycoside low-,medium-and high-dose groups(200,500,1 000 µmol·L-1).Cell viability was detected by CCK-8 method.The apoptosis rate was detected by flow cytometry.The mRNA expression levels of CYP1A1,CYP3A4 and Caspase-3 in hepatocytes were detected by qRT-PCR.The expression levels of CYP1A1,CYP3A4 and Cleaved Caspase-3 in hepatocytes were detected by Western Blot.(2)In vivo experiment:Kunming mice were randomly divided into blank group,Polygoni Multiflori Radix group(8 g·kg-1),prepared Polygoni Multiflori Radix group(8 g·kg-1),emodin group(37.8 mg·kg-1),rhein group(312 µg·kg-1)and gallic acid group(4.58 g·kg-1),with 10 mice in each group.Intragastric administration was performed once a day for 4 weeks.The liver function indexes of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),total bilirubin(TBIL)and indirect bilirubin(IBIL)were detected by automatic microplate reader.The pathological changes of liver tissue were observed by HE staining.The mRNA expression levels of CYP1A1,CYP3A4 and Caspase-3 m in liver tissue were detected by qRT-PCR.The expression levels of CYP1A1,CYP3A4 and Cleaved Caspase-3 in liver tissue were detected by Western Blot.Results(1)In vitro experiments:Compared with the blank group,medium-and high-doses of emodin,as well as different concentrations of rhein and gallic acid,significantly inhibited HepG2 cells(P＜0.01),showing concentration dependence.Stilbene glycoside at all concentrations showed no significant inhibitory effect on HepG2 cells(P＞0.05).Compared with the blank group,the morphology of cells in all treatment groups changed with increasing drug concentration,showing shrinkage,rounding,darkening,and partial necrosis and detachment.Severe necrosis was observed in the medium-and high-dose emodin groups and the medium-and high-dose gallic acid groups.The apoptosis rate significantly increased in the medium-and high-dose emodin,rhein,and gallic acid groups(P＜0.05,P＜0.01).The mRNA expression of CYP1A1 was significantly upregulated in all dose groups of emodin,rhein,and gallic acid(P＜0.05,P＜0.01).The mRNA expression of CYP3A4 was significantly upregulated in the high-dose emodin group,medium-and high-dose rhein groups,and all dose groups of gallic acid(P＜0.01).The mRNA expression of Caspase-3 was significantly upregulated in the medium-and high-dose emodin groups,high-dose rhein group,and medium-and high-dose gallic acid groups(P＜0.05,P＜0.01).The protein expressions of CYP1A1,CYP3A4,and Cleaved Caspase-3 were significantly upregulated in all dose groups of emodin,rhein,and gallic acid(P＜0.05,P＜0.01).(2)In vivo experiments:Based on the degree of liver tissue pathological damage,the order was roughly gallic acid group＞rhein group＞emodin group＞Polygoni Multiflori Radix group＞prepared Polygoni Multiflori Radix group,with the prepared Polygoni Multiflori Radix group showing the least liver tissue damage.Compared with the blank group,the serum levels of IBIL,TBIL,AST,and ALP were significantly increased in the Polygoni Multiflori Radix group,emodin group,and gallic acid group(P＜0.05,P＜0.01).The serum levels of AST and ALP were significantly increased in the rhein group(P＜0.05,P＜0.01),and the serum ALT level was significantly increased in the gallic acid group(P＜0.05).The mRNA expressions of CYP1A1,CYP3A4,and Caspase-3 in liver tissues were significantly upregulated in the Polygoni Multiflori Radix group,prepared Polygoni Multiflori Radix group,emodin group,and gallic acid group(P＜0.05,P＜0.01),and the mRNA expression of CYP3A4 was significantly upregulated in the rhein group(P＜0.05).The protein expressions of CYP1A1 and CYP3A4 wrere significantly upregulated in the Polygoni Multiflori Radixgroup,prepared Polygoni Multiflori Radix group,and gallic acid group(P＜0.05,P＜0.01),and the protein expression of Cleaved Caspase-3 was significantly upregulated in the Polygoni Multiflori Radix group,rhein group,and gallic acid group(P＜0.05,P＜0.01).Conclusion The mechanism of liver injury induced by Polygoni Multiflori Radix and its main components may be related to the activation of CYP1A1 and CYP3A4 pathways and the promotion of Caspase-3 apoptosis factor expression,among which gallic acid has the strongest liver injury effect.关键词
何首乌/制首乌/大黄素/大黄酸/没食子酸/药物性肝损伤/CYP酶/Caspase-3途径/HepG2细胞/小鼠Key words
Polygoni Multiflori Radix/prepared Polygoni Multiflori Radix/emodin/rhein/gallic acid/drug-induced liver injury/CYP enzyme/Caspase-3 pathway/HepG2 cells/mice分类
医药卫生引用本文复制引用
刘思雨,高峰,张依娜,李敏,雷一鸣,王丽平,卫培峰,欧莉,白杨,董泰玮..基于CYP酶/Caspase-3途径探讨何首乌及其主要成分致药物性肝损伤的作用机制[J].中药新药与临床药理,2025,36(4):506-514,9.基金项目
陕西省科技厅重点项目(2021JQ-728) （2021JQ-728）
中医药"双链融合"中青年科研创新团队建设项目(2022-SLRH-YQ-008) （2022-SLRH-YQ-008）
陕西省"特支计划"人才项目. （）
