商陆抗病毒蛋白在急性人乙型肝炎病毒感染小鼠体内的抑制作用

Inhibition of pokeweed antiviral protein on hepatitis B virus replication in acute human hepatitis B virus infected mice

目的探讨商陆抗病毒蛋白（PAP）在急性HBV感染小鼠模型体内抗HBV的效果。方法通过尾静脉注射具有复制能力的HBV真核表达质粒来建立小鼠急性HBV感染模型，根据注射后第1天小鼠血清HBsAg和HBeAg水平，从35只小鼠中选出24只进行配对，分为PAP治疗组（腹腔注射0．25mg／kgPAP）和对照组。于PAP注射前、注射后第1、3、5、7天，时问分辨免疫荧光分析法检测小融血清HBsAg、HBeAg和抗HBc水平，荧光定量PCR检测小鼠血清HBVDNA水平。注射后第7天HE染色检测肝组织病理变化，免疫组织化学检测肝组织HBsAg、HBeAg表达。采用配对t检验进行统计学分析。结果与对照组相比，在PAP腹腔注射后第1、3、5天对HBsAg的抑制率分别为23％（t=116．3，P〈O．05）、47％（t=38．2，P〈0．05）、68％（t=23．7，P〈0．05），对HBeAg的抑制率分别为36％（t=34．2，P〈0．05）、55％（t=61．6，P〈0．05）、83％（t=98．8，P〈0．05），对HBVDNA的抑制率分别为70．7％（t=6．6，P〈0．05）、86．9％（t=5．9，P〈0．05）、95．2％（t=36．6，P〈0．05）、95．3％（t=19．7，P〈0．05）。结论0．25mg／kg剂量的PAP在急性HBV感染小鼠模型体内对血清和肝组织的HBsAg、HBeAg的表达及HBVDNA的复制均有明显的抑制效果。

Objective To explore the antiviral effects of pokeweed antiviral protein （PAP） in mouse model of acute hepatitis B virus （HBV） infection. Methods Mouse model of acute HBV infection was established by hydrodynamictail vein injection with eukaryotic expression plasmid containing replicable HBV. Based on serum levels of hepatitis B surface antigen （HBsAg） and hepatitis B e antigen （HBeAg） at day 1 after injection, 24 mice were selected from 35 mice and averagely divided into PAP treated group （intraperitoneally injected with 0. 25 mg/kg PAP） and control group. Before injection and 1, 3, 5, 7 days after PAP injection, serum levels of HBsAg, HBeAg and anti-HBc were detected by enzyme-linked immunosorbent assay （ELISA）. Serum level of HBV DNA was determined by fluorescence quantitative PCR （FQ-PCR）. After 7 days of PAP injection, liver histopathological changes were examined using HE staining and expressions of HBsAg and HBeAg in the liver were assayed by immunohistochemical staining. The data were analyzed by paired t test. Results Compared with control group, the inhibition rates of HBsAg at dayl, 3, 5 after PAP intraperitoneal injection were 23 %, 47 % and 68 %, respectively （t = 116.3, t = 38.2, t = 23.7 ; all P〈0.05）, those of HBeAg were 36 %, 55% and 83%, respectively （t=34.2,t=61.6,t=98.9; all P〈0.05）, those of HBV DNA were 70.7%, 86.9%, 95.2% and 95.3%, respectively （t=6.6, t=5.9,t=36.6,t=19.7;all P〈0. 05）. Conclusion PAP with dose of 0. 25 mg/kg may significantly inhibit expressions of HBeAg and HBeAg and replication of HBV DNA in serum and liver tissues in acute HBV infected mice.