扁塑藤素对HepG2.2.15细胞生长的影响及其对乙肝病毒的抑制作用  

Effect of pristimerin on the growth of HepG2. 2. 15 and inhibition of the hepatitis B virus replication

作　　者：张欣蕊[1] 易学瑞 孔祥平 ZHANG Xinrui;YI Xuerui;KONG Xiangping(Department of Infection and Immunity,the Second Hospital of Tianjin Medical University,Tianjin,300211 P.R. China;The Air Force Hospital of Southern Theater Command,Guangzhou,Guangdong,510600 P.R. China)

机构地区：[1]天津医科大学第二医院感染免疫科,天津300211 [2]南部战区空军医院,广东广州510600

出　　处：《华西药学杂志》2019年第5期444-447,共4页West China Journal of Pharmaceutical Sciences

基　　金：国家科技重大专项(2012ZX10004503-010);广东省科技计划资助项目(2013B060300015)

摘　　要：目的研究扁塑藤素在体外对肝癌细胞株HepG2. 2. 15细胞生长的影响及对乙肝病毒(HBV)的抑制作用。方法将不同浓度的扁塑藤素作用于转染HBV全基因组DNA的肝癌细胞株HepG2. 2. 15,作用24 h后收集上清,采用MTT法测定扁塑藤素对HepG2. 2. 15细胞生长的影响,通过形态学观察扁塑藤素对HepG2. 2. 15细胞生长的影响;采用改良的两步灌流法分离HBV转基因小鼠原代肝细胞;采用MTT法观察不同浓度扁塑藤素对原代肝细胞的毒性;应用ELISA法和PCR法测定乙肝表面抗原(HBsAg)和HBV-DNA含量。结果扁塑藤素可以影响HepG2. 2. 15细胞的生长,显微镜下亦可见被药物作用后的细胞脱落明显,脱落细胞悬浮于培养液中,形态变圆,折光性差;扁塑藤素对原代肝细胞的最大无毒浓度为2μmol·L^-1,最大无毒浓度以下,浓度为2、0. 2、0. 02μmol·L^-1的扁塑藤素可以有效抑制原代肝细胞HBsAg和HBV-DNA的表达,且随着扁塑藤素浓度的减低对HBsAg和HBV-DNA的抑制作用减弱。结论扁塑藤素可以影响HepG2. 2. 15细胞的生长;在HBV转基因小鼠原代肝细胞中,一定浓度的扁塑藤素可以抑制其HBsAg和HBV-DNA的表达。OBJECTIVE To investigate the effect of pristimerin on the growth of hepatoma cell line HepG2. 2. 15 and inhibition of the hepatitis B virus( HBV) replication. METHODS We used the HepG2. 2. 15 cells,which were treated with different concentrations of pristimerin for 24 hours. MTT assay was used to detect the effects of cell growth,and cell morphology was observed by microscope. We separated the primary hepatocytes from HBV transgenic( HBV-Tg) mice with the modified two-steps method. The cytotoxicity of pristimerin on the primary hepatocytes was analyzed also by MTT assay. The changes of hepatitis B surface antigen( HBs Ag) and HBV-DNA in the supernatant were determined with ELISA and PCR. RESULTS We found pristimerin can influence the growth of HepG2. 2. 15 cells. Microscope showed that the cells were dropped significantly after treated with pristimerin,and suspended in the supernatant with round shape and poor refraction. The maximum avirulent concentration of pristimerin on the primary hepatocytes was 2 μmol·L^-1. HBs Ag and HBV-DNA of the cells treated with pristimerin was effectively inhibited at the concentration of 2-0. 02 μmol·L^-1. As the concentration of pristimerin was reduced,there was a corresponding reduce in inhibition in HBs Ag and HBV-DNA. CONCLUSION A certain concentration of pristimerin can not only influence the growth of HepG2. 2. 15 cells,but also can inhibit the expression of the HBs Ag and HBV-DNA in the primary hepatocytes.

关 键 词：扁塑藤素 HepG2. 2. 15 乙型肝炎病毒转基因小鼠原代肝细胞 乙型肝炎病毒 原发性肝癌 蛋白酶体抑制剂 乙肝表面抗原 乙型肝炎病毒-DNA 

分 类 号：R96[医药卫生—药理学]