紫花地丁抗乙型肝炎病毒的实验研究  

Anti-HBV effect of Viola yedoensis Makino in vitro and in vivo

作　　者：王玉[1] 吴中明[1] 敖弟书[1] 

机构地区：[1]遵义医学院,贵州563003

出　　处：《中药药理与临床》2011年第5期70-74,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：贵州省硕士研究经费;贵州省遵义医学院基金项目(No F-339)

摘　　要：目的:观察紫花地丁水浸出物抗乙型肝炎病毒的作用。方法:以HepG2.2.15细胞作为体外实验模型,50μg/ml拉米夫定(3TC)和不加药培养液作为阳性、阴性对照,给予含有0.12、0.6、3mg/ml紫花地丁浓度的培养液处理3、6、9天后,通过MTT法观察药物对细胞生长的影响;采用Real-time PCR法和ELISA法检测细胞上清液中HBV DNA和HBsAg、HBeAg的含量。体内试验选1日龄麻鸭,人工感染DHBV后随机分为7组:紫花地丁水浸出物1.2、6、30mg/kg.d,3TC 1mg/kg.d和紫花地丁水浸出物1.2mg/kg.d联合用药,对照组采用生理盐水和2mg/kg.d 3TC。每组5只,每天灌胃给药1次,连续10天,观察药物对DHBV的作用。结果:紫花地丁所有实验浓度对HepG2.2.15细胞均无毒性,且对HBsAg、HBeAg都有一定的抑制作用。其中0.6mg/ml组对HBsAg的抑制作用随着培养时间延长逐渐增强,第9天达到35.4%。3mg/ml组对HBeAg的抑制率在第6天最高,达到38.6%。紫花地丁水浸出物能有效地抑制HBV DNA复制,最高抑制率达到86.1%;3mg/ml组在第9天抑制率仍有42.3%,明显高于3TC(24.2%)。在体内试验中发现6mg/kg.d紫花地丁水浸出物具有抑制DHBV复制作用。结论:紫花地丁在体内、外试验中均有抗HBV活性作用。Objective ： To study the anti-Hepatitis B virus effect of Viola yedoensis Makino.Methods ： HepG2.2.15 cells as study model were treated with aqueous extract of Viola yedoensis Makino that were diluted into 0.12 mg/ml,0.6 mg/ml and 3mg/ml groups for 3 days,6 days and 9 days.The experiment set up positive control group（3TC,50μg/ml） and normal control group.The cell toxicity of Viola yedoensis Makino was determined with MTT method,and the effects of Viola yedoensis Makino on HBsAg and HBeAg extracted by HepG2.2.15 cells were determined with ELISA method.Real-time fluorescence quantitative PCR was used for determining extracted HBV DNA in the culture medium.One-day old ducks infected DHBV were randomly divided into seven groups： different dose concentrations of aqueous extract of Viola yedoensis Makino（1.2 mg/kg/d、6 mg/kg/d and 30mg/kg/d）;drug combination group of 3TC 1mg/kg/d and aqueous extract of Viola yedoensis Makino 1.2mg/kg/d;positive control drug： 3TC 2 mg/kg/d;virus control group and blank control group were administered by normal saline.Five animals were used in each group.Drugs were intragastric administration,once daily for 10 days,then the influence of drugs on DHBV-DNA on ducks was observed.Results ： All concentrations of aqueous extract of Viola yedoensis Makino had no cytotoxicity and could inhibit HBsAg and HBeAg.0.6mg/ml group of aqueous extract of Viola yedoensis Makino had inhibitory effect on HBsAg in time-depentent manner,and the inhibitory rates reached the peak of 35.4% at 9th day;3mg/ml group attained the highest inhibitory rates at 38.6% after 6 days.Compared to the positive group,aqueous extract of Viola yedoensis Makino had a significant inhibition effect on HBV DNA,the highest inhibitory rates are 86.1%.But 3mg/ml group had stronger inhibition effect than 3TC at 9th day,the inhibition rate was 42.3%.The middle-dose concentration of aqueous extract of Viola yedoensis Makino shows apparent anti-DHBV effect in vivo.Conclusion： Viola yedoensis Makino has effective anti-

关 键 词：紫花地丁 乙型肝炎病毒 体外实验 体内试验 

分 类 号：R285[医药卫生—中药学]