全蝎和蜈蚣的多肽粗提物对肝癌细胞中HBV的抑制作用  

Inhibition of Hepatitis B virus in liver cancer cells by preliminary peptide extracts of scorpion and centipede

作　　者：马青[1] 杨扬 马韬[3] 徐卫东[1] 赵明[1] MA Qing;YANG Yang;MA Tao;XU Weidong;ZHAO Ming(Pharmacy College,Jiangsu University,Zhenjiang Jiangsu 212013;Shanghai Institute of Planned Parenthood Research,Shanghai 200032;Department of General Surgery,Ruijin Hospital Affiliated to Shanghai Jiao Tong University,Shanghai 200025,China)

机构地区：[1]江苏大学药学院,江苏镇江212013 [2]上海计划生育科学研究所,上海200032 [3]上海交通大学附属瑞金医院普外科,上海200025

出　　处：《江苏大学学报（医学版）》2021年第5期431-437,共7页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81573815);江苏省2019年研究生创新计划(SJCX19-0575)。

摘　　要：目的:研究全蝎尾部多肽粗提物(peptide extract from scorpion tail,PEST)与蜈蚣头部多肽粗提物(peptide extract from centipede head,PECH)对肝癌细胞乙型肝炎病毒(Hepatitis B virus,HBV)合成的影响及其可能机制。方法:超滤法提取PEST与PECH。选取肝癌HepAD38和HepG2细胞,将其分组:对照组,用含10%胎牛血清的培养基培养;PEST组,不同浓度PEST(0.125、0.250、0.500和1.000 mg/mL)处理;PECH组,不同浓度PECH(0.125、0.250、0.500和1.000 mg/mL)处理;阳性对照组,拉米夫定或恩替卡韦或四环素处理。MTT法检测细胞活力,qRT-PCR检测HepAD38细胞上清液HBV DNA含量。另取HepAD38细胞,分为PEST组(1 mg/mL PEST处理)、PECH组(1 mg/mL PECH处理)、对照组、阳性对照组和阴性对照组;ELISA法检测细胞上清液中HBsAg和HBeAg含量,qRT-PCR检测HBV X、S、preC、P mRNA表达量,蛋白印迹法检测HBV核心蛋白(HBV core protein,HBc)表达。结果:PEST或PECH处理的肝癌细胞HepG2和HepAD38细胞活力均在70%以上;与对照组相比,PEST组(0.250、0.500和1.000 mg/mL)和PECH组(0.125、0.500和1.000 mg/mL)HBV DNA拷贝数明显降低(P均<0.05);与对照组相比,PEST组和PECH组HBsAg和HBeAg含量明显降低(P均<0.05),HBV P mRNA相对表达明显降低(P均<0.05),PEST组HBc表达几乎无差异,而PECH组HBc表达减少。结论:PEST和PECH在HepAD38细胞株中表现出抗HBV作用,可能与其抑制HBV P mRNA合成有关。Objective:To explore the effects of scorpion peptide preliminary extract(peptide extract from scorpion tail,PEST)and the centipede preliminary peptide extract(peptide extract from centipede head,PECH)on Hepatitis B virus synthesis and potential mechanism in liver cancer cells.Methods:Ultrafiltration was used to extract PEST and PECH.HepAD38 and HepG2 cells were selected and divided into control group,cells were cultured with high sugar medium containing with 10%fetal bovine serum;PEST group,cells were treated with different concentrations(0.125,0.250,0.500 and 1.000 mg/mL)of PEST;PECH group,cells were treated with different concentrations(0.125,0.250,0.500,1.000 mg/mL)of PECH;positive control group,cells were treated with lamivudine or entecavir or tetracycline;MTT assay was used to detect cell viability.The qRT-PCR was used to detect Hepatitis B virus DNA content in the supernatant of HepAD38 cells.In addition,HepAD38 cells were divided into PEST group(cells were treated with 1 mg/mL PEST),PECH group(cells were treated with 1 mg/mL PECH),control group,positive control group and negative group;HBsAg and HBeAg content were detected by ELISA;HBV X,S,preC,P mRNA relative expression level were detected by qRT-PCR;Hepatitis B virus core protein synthesis was detected by Western blotting.Results:The cell viability of HepG2 and HepAD38 cells treated with PEST or PECH were both above 70%;compared with the control group,PEST(0.250,0.500 and 1.000 mg/mL)or PECH(0.125,0.500 and 1.000 mg/mL)group showed significantly decreased Hepatitis B virus DNA copy number(all P<0.05);compared with the control group,PEST or PECH group showed significantly lower content of HBsAg and HBeAg and expression level of Hepatitis B virus P mRNA(all P<0.05),PEST group had almost no effect on core protein expression,while PECH group decreased Hepatitis B virus core protein expression.Conclusion:PEST and PECH exerted anti-Hepatitis B virus effect on HepAD38 cell,which may be related to its inhibition on P mRNA synthesis of Hepatitis B virus.

关 键 词：乙型肝炎病毒 全蝎 蜈蚣 多肽 HepAD38细胞 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]