苦参碱调控PXR-CYP3A4通路对石胆酸诱导肝损伤细胞系的保护作用  

Protective Effect of Matrine on Lithocholic Acid Induced Liver Injury Cell Lines by Regulating PXR-CYP3A4 Pathway

作　　者：阳丽梅[1] 陈淑芳 黄旭慧[2] 庄捷[1] YANG Li-mei;CHEN Shu-fang;HUANG Xu-hui;ZHUANG Jie(Department of Pharmacy,Fujian Prorincial Hospital,Provincial Clinical Medical College of Fujian Medical Uniwersity,Fuzhou 350000,China;Department of Pharmacy,Fujian Provincial Hospital South Branch,Frujian Provincial Hospital Jinshan Branch,Fuzhou 350000,China)

机构地区：[1]福建医科大学省立临床医学院,福建省立医院药学部,福州350000 [2]福建省立医院南院,福建省立金山医院药学部,福州350000

出　　处：《中国药学杂志》2020年第21期1802-1806,共5页Chinese Pharmaceutical Journal

基　　金：福建省医学创新课题资助(2016-CX-15);福建省卫生健康中青年骨干人才培养项目资助(2019-ZQN-34)。

摘　　要：目的探索苦参碱(MT)对石胆酸(LCA)诱导肝损伤细胞系的保护作用及其可能的作用机制。方法选择人张氏肝细胞(Changlivercell)和Huh-7细胞,分别采用含双抗的1640培养基和DMEM培养基培养。将细胞分成空白组、LCA组和不同剂量MT(0.5、1和2μmol·L^-1)组,分别给药后,全自动生化分析仪检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBiL)和碱性磷酸酶(ALP),RT-PCR法检测孕烷X受体(PXR)和CYP3A4 mRNA表达,Western blot法检测PXR和CYP3A4蛋白的表达。结果对于人张氏肝细胞,MT可以显著降低LCA干预后细胞ALT、AST、TBiL和ALP水平;上调细胞的PXRmRNA表达,而下调CYP3A4mRNA表达;下调细胞PXR和CYP3A4蛋白的表达。对于Huh-7细胞,MT可显著降低LCA干预后细胞的ALT、AST和ALP水平;上调PXR和CYP3A4 mRNA表达;下调CYP3A4蛋白的表达,而对PXR蛋白表达无明显影响。结论MT可改善LCA诱导肝损伤细胞的肝功能指标,其机制可能与调控PXR-CYP3A4通路mRNA和蛋白表达有关,其对Changlive细胞和Huh-7细胞的影响不尽相同。OBJECTIVE To objective of this study was to explore the protective effect and potential mechanism of matrine(MT) on lithocholine acid(LCA)-induced liver injury Chang-liver and Huh-7 cell lines. METHODS Chang liver cell line and Huh-7 cell lines were respectively cultured in 1640 medium and DMEM medium which contained penicillin and streptomycin. Four groups which were blank group, LCA group, and different doses of MT group(0.5, 1, 2 μmol·L^-1) were installed. After intervention with agents, ALT, AST, TBIL, ALP of cells were detected by automatic biochemical analyzer. The expression of PXR and CYP3 A4 mRNA was detected by RT-PCR, and the expression of PXR and CYP3 A4 protein was detected by Western blot. RESULTS For Chang liver cell, MT significantly decreased ALT, AST, TBiL and ALP levels, up-regulated PXR mRNA expression, down-regulated CYP3 A4 mRNA expression, and down-regulated PXR and CYP3 A4 protein expression in cells which were treated by LCA. For Huh-7 cell, MT significantly decreased ALT, AST and ALP levels, up-regulated PXR and CYP3 A4 mRNA expression and and down-regulated CYP3 A4 protein expression in cells which were treated by LCA. CONCLUSION This study shows that MT could improve the liver function index in LCA-induced injury liver cells. The mechanism might be related to the regulation of mRNA and protein expression of PXR and CYP3 A4. However, the effects of MT on Chang live cells and Huh-7 cells are different.

关 键 词：苦参碱 肝损伤 人张氏肝细胞系 HUH-7细胞 孕烷X受体 CYP3A4 

分 类 号：R965[医药卫生—药理学]