复方虎杖方体内外抗乙肝病毒的活性评价  

Evaluation of the Anti-Hepatitis B Virus Activity of Compound Huzhang Prescription in vivo and in vitro

作　　者：王云[1] 鞠建明[1] 李振华[1] 刘克冕[1] 汪露露 彭蕴茹[1] 刘超[1] WANG Yun;JU Jian-ming;LI Zhen-hua;LIU Ke-mian;WANG Lu-lu;PENG Yun-ru;LIU Chao(The Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing,210028,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏省中医药研究院,江苏南京210028

出　　处：《南京中医药大学学报》2021年第1期62-66,共5页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：江苏省“六大人才高峰”高层次人才项目(2015-YY-011);江苏省“333高层次人才培养工程”科研资助项目(BRA2015494);江苏省研究生培养创新工程研究生科研与实践创新计划项目(SJCX20_0571)。

摘　　要：目的研究复方虎杖方体内外抗乙肝病毒作用。方法体外实验采用CCK-8染色法检测复方虎杖方水提物、醇提物对HepG2.2.15细胞株的抗增殖活性,采用化学发光微粒子免疫检测法测定HBsAg、HBeAg含量,采用实时荧光定量PCR法检测HBV-DNA含量;体内实验采用血清斑点杂交法(Dot blot)测定给药5、10 d后及停药3 d后鸭血清中的DHBV-DNA含量。结果体外结果显示,复方虎杖方水提物、醇提物能有效抑制HepG2.2.15细胞增殖活性,IC 50分别为51.28 mg/mL和46.78 mg/mL;复方虎杖方能有效抑制HepG2.2.15细胞分泌HBsAg、HBeAg,具有明显的剂量依赖关系,醇提物对HBeAg分泌的抑制作用明显强于水提物;复方虎杖方醇提物能有效抑制HepG2.2.15细胞HBV-DNA复制,但水提物效果不明显。体内结果显示,复方虎杖方能有效抑制鸭血清中DHBV-DNA复制,且能显著抑制停药后的病毒反跳。结论复方虎杖方体内和体外均有一定的抗乙肝病毒作用,研究为该方的开发应用奠定了基础。OBJECTIVE To investigate the anti-hepatitis B virus(HBV)effect of Compound Huzhang Prescription in vivo and in vitro.METHODS In vitro,CCK-8 staining method was used to determine the anti-proliferative activity of Compound Huzhang Prescription against HepG2.2.15 cell lines expressing HBV,chemiluminescent microparticle immunoassay method was used to determine hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)levels,and fluorogenic quantitative PCR method was used to determine HBV-DNA level.In vivo,Dot blot assay was used to investigate the content of DHBV-DNA in serum at the 5th,10th day after administration and the 3th day after drug withdrawal.RESULTS The results showed that water extract and alcohol extract of Compound Huzhang Prescription had a good inhibitory effect on the growth of HepG2.2.15 cells,with a half inhibitory concentration(IC 50)of 51.28 mg/mL and 46.78 mg/mL,respectively.Compound Huzhang Prescription significantly inhibited HBsAg and HBeAg expressions and it showed a significant dose effect relationship.The inhibition of alcohol extract was stronger than that of water extract.The alcohol extract had an inhibitory effect on HBV-DNA replication,but the effect of water extract was not obvious.Compound Huzhang Prescription effectively inhibited DHBV-DNA replication,and significantly inhibited the virus rebound after drug withdrawal in vivo.CONCLUSION Compound Huzhang Prescription shows a certain anti-hepatitis B virus effect in vivo and in vitro,which lays a foundation for the development and application.

关 键 词：复方虎杖方 HEPG2.2.15细胞 鸭乙型肝炎病毒 HBsAg HBEAG 抗乙肝病毒作用 

分 类 号：R285.5[医药卫生—中药学]