康赛宁对体外培养的人肝癌细胞的影响及与乙肝疫苗联合免疫对小鼠的免疫增强作用

Influence of Kanserin on Human Hepatoma Cells Cultured In Vitro and Enhancement of Immunity of Mice by Combined Vaccination with Kanserin and Hepatitis B Vaccine

目的研究A群溶血性链球菌制剂(康赛宁)对体外培养的人肝癌细胞系Hep3B和HepG22.2.15的影响及与乙肝疫苗联合免疫对小鼠的免疫增强作用。方法用不同浓度的康赛宁作用于人肝癌细胞Hep3B和HepG22.2.15,观察细胞形态学,MTT比色法检测细胞活性,EIA法检测培养上清HBsAg、HBeAg含量,吖啶橙直接染色检测细胞凋亡,Southernblot分析细胞DNA片段。给NIH小鼠腹腔注射康赛宁和乙肝疫苗,检测血清中HBs抗体、转氨酶水平。给普通小鼠腹腔注射康赛宁和乙肝疫苗,观察注射前后的体重变化。结果康赛宁可抑制HBsAg及HBeAg的分泌,使染色体DNA发生有规律的断裂,促使细胞发生凋亡。与乙肝疫苗联合免疫,可提高小鼠血清中HBs抗体水平。无论单用或合用康赛宁,对小鼠均无明显毒副作用。结论康赛宁对体外培养的人肝癌细胞系有影响,与乙肝疫苗联合免疫对小鼠有免疫增强作用。

Objective To study the influence of Kanserin, a preparation of group A hemoloytic streptococci, on the human heparoma cells cultured in vitro and the enhancement of immunity of mice by combined vaccination with Kansefin and hepatitis B vaccine. Methods Add the Kansefin at various concentrations into the human hepatoma cells Hep3B and HepG22. 2.2. 15 cultured in vitro, observe the cell morphology and determine the cell activity by MTT method. Analyze the HBaAg and HBeAg contents in Culture supernatant by EIA,the cell DNA fragments by Southern blot and the cell apoptosis by direct aetinide orange staining. Inject i.p. NIH mice with Kanserin and hepatitis B vaccine and determine the anti-HBs antibody and transaminase levels in sera. Inject i.p. common mice with Kanserin and hepatitis B vaccine and observe the change of bodyweight before and after injection. Resdts Kanserin inhibited the secretion of HBsAg and HBeAg, caused the regular chromosome DNA fragmentation and leaded the cells to apoptosis. The combined vaccination with Kanserin and hepatitis B vaccine increased the anti-HBs level in murine sera. Neither combined vaccination with Kanserin and hepatitis B vaccine nor vaccination with hepatitis B alone caused adverse reactions. Conclusion Kanserin showed significant influence on human hepatoma cells cultured in vitro. The combined vaccination with Kanserin and hepatitis B vaccine enhanced the immunity of mice.