蟾酥缓释注射液的免疫佐剂活性

Adjuvant effect of sustained-release injection of Venenum bufonis

为考察蟾酥缓释注射液体外释药行为,并研究其耳根穴注射免疫佐剂作用。本试验采用紫外分光光度法测定蟾酥缓释注射液在无膜溶出模型中的体外释放度;ICR小鼠皮下注射卵清白蛋白(OVA),同时耳根穴注射蟾酥缓释注射液,二免2周后分离血清和脾细胞,ELISA法检测血清OVA特异性抗体IgG水平,MTT法检测脾淋巴细胞刺激指数。结果:药物体外释放行为遵循零级动力学方程,其累积药物释放百分率(Q)与时间(t)的回归方程为Q=3.989t+0.322(r=0.999);蟾酥缓释注射液和蟾酥注射液耳根穴注射后血清OVA特异性抗体IgG水平和淋巴细胞刺激指数均显著高于正常免疫组(P<0.05),其中蟾酥缓释注射液组优于蟾酥注射液组,并减少了注射次数。结论蟾酥缓释注射液具有缓释作用,可以通过穴位注射提高抗原免疫水平。

To investigate the in vitro drug release behavior of the sustained-release injection of Venenum Bufonis and evaluate its adjuvant effect by ear acupoint injection.UV spectrophotometry was established to determine the release rate of the sustained-release injection of Venenum Bufonis in the memberless model.ICR mice were immunized subcustaneously twice with ovalbumin（OVA） and injected with the sustained-release injection of Venenum Bufonis at ear acupoint.Two weeks after the boosting,OVA-specific IgG antibody levels in serum and splenocyte proliferation were determined.The drug release behavior followed the zero order kinetics and cumulative drug release（Q） equations was Q=3.989t＋0.322（r=0.999）.Serum OVA-specific IgG and splenocyte stimulation indexes were significantly enhanced by the sustained-release injection of Venenum Bufonis and Venenum Bufonis injection compared with the control immune group（P〈0.05）,while the adjuvant potential of the sustained-release injection of Venenum Bufonis was better with lower injection times.The sustained-release injection of Venenum Bufonis can enhance the antigenic immune response by acupoint injection.