商陆抗病毒蛋白抑制乙型肝炎病毒复制的分析-analysis of pokeweed antiviral protein inhibiting hepatitis b virus replication.docx

华 华 中 科 技 大 学 博 士 学 位 论 文 PAGE 14 PAGE 14 免疫组化染色显示 PAP12 组及 PAP12-HBc 融合蛋白组小鼠肝脏中 HBsAg 阳性细胞数 量较生理盐水组减少。PAP12 组、PAP12-HBc 融合蛋白组的 HBV mRNA 较生理盐水组 少。 结论：PAP12 及 PAP12-HBc 融合蛋白在慢性 HBV 感染小鼠体内对 HBV 有抑制作 用，并未见明显肝肾毒性。 关键词： PAP；HBc；乙型肝炎病毒；抗病毒； The Inhibition of Hepatitis B virus replication by pokeweed antiviral protein Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology Doctor Candidate: Chen Xiliu Supervisor: Prof.He Yongwen ABSTRACT Part Ⅰ Preparation of the pokeweed antiviral protein from seeds and assay of its toxicity Objective: To purified the pokeweed antiviral protein from seeds (PAP-S) from Phytolacca Americana and test its acute and subacute toxicity in mice. Methods: The PAP-S was purified from the seeds of Phytolacca Americana using ion exchange chromatography CM 50 and CM 52 and molecular sieve G 50. The molecular weight was tested by SDS/polyacrylamide-gel electrophoresis (SDS) and the concentration of protein was checked by BCA Protein Assay Kit. 80 male Kunming mice were divided into 8 groups randomly. In each group, PAP-S dissolved in 100 μl of saline was injected intraperitoneally once at different doses ranging from 5.6 to 0.5 mg/kg of body weight, with a ratio between doses of 1.4. The health condition and mortality of mice was observed for 14 days after injection. 40 mice were assigned to 4 groups randomly. PAP-S was injected intraperitoneally at different doses of 0, 0.125, 0.25, 0.5 mg/kg of body weight per day for 14 days. 24 hours after the last administration the mice were euthanized. Sera samples were collected to test the levels of alanine aminotransferase (ALT) and blood urea nitrogen (BUN). The tissue of liver and kidney were taken to be stained with haematoxylin and eosin for histological analysis. Results: The effluent representing PAP-S was freeze-dried to protein powder approximate to 200 mg and demonstrated a single 30-kDa band in SDS. The LD50 values of PAP-