哈蟆油的免疫调节作用及其微球的制备工艺

Immune regulation of oviductus Ranae and preparation of its microspheres

目的:研究哈蟆油对免疫系统的影响,并探讨其制备成微球的工艺。方法:50只ICR小鼠随机分为空白对照组、模型组、哈蟆油低剂量组(0.1g.kg-1)、哈蟆油中剂量组(0.4g.kg-1)和哈蟆油高剂量组(1.0g.kg-1),每组10只。采用流式细胞术检测小鼠脾T淋巴细胞亚群含量,ELISA法测定脾T淋巴细胞中白细胞介素2(IL-2)含量。以喷雾干燥法制备哈蟆油微球,采用正交试验法考察进风温度、出风温度、物料比、进料速度,以平均粒径、粒径分布、载药量、包封率为考察指标,确定哈蟆油微球最佳制备工艺。结果:与空白对照组比较,模型组小鼠的脾T淋巴细胞亚群CD3+、CD4+水平和CD4+/CD8+比值明显增加,差异均有统计学意义(P<0.001);与模型组比较,哈蟆油低剂量组小鼠的脾T淋巴细胞亚群CD3+和CD4+水平明显降低(P<0.001),CD4+/CD8+比值明显降低(P<0.05);与空白对照组比较,模型组小鼠的脾T淋巴细胞中IL-2含量明显降低(P<0.001);与模型组比较,哈蟆油低、中、高剂量组小鼠的脾T淋巴细胞中IL-2含量均显著增高(P<0.001)。采用喷雾干燥法制备微球,最佳工艺为A3B1C1D1,即物料比1∶3、进风温度130℃、出风温度100℃、进料速度30mL.min-1。结论:哈蟆油具有调节免疫力的作用;将其制备成微球,工艺简单合理、适合大工业生产。

Objective To study the effects of oviductus ranae on the immune system and prepare its microspheres.Methods Fifty ICR mices were randomly divided into control group,model group,low dose oviductus ranae group（0.1 g·kg^-1）,middle dose oviductus ranae group（0.4 g·kg^-1）and high dose oviductus ranae group（1.0 g·kg^-1）,the contents of spleen T lymphocyte subpopulations in mice were determined by flow cytometry,and the content of IL-2 in spleen T lymphocytes was detected by ELISA.The microspheres of oviductus ranae were prepared by spray drying,orthogonal experiment was used to investigate the material ratio,inlet temperature,outlet temperature,feed flow rate with average particle size,range of particle size,drug loading and the degree of encapsulated efficiency,the optimum preparation of oviductus ranae microspheres was determined.Results Compared with control group,the contents of spleen T lymphocyte subpopulations CD3^＋,CD4^＋ levels and CD4^＋/CD8^＋ ratio in the mice in model group were significantly increased（P〈0.001）;Compared with model group,the contents of spleen T lymphocyte subpopulations CD3^＋,CD4^＋ levels in the mice in low dose oviductus ranae group were decreased significantly（P〈0.001）,CD4^＋/CD8^＋ ratio was also decreased significantly（P〈0.05）.Compared with control group,the content of IL^-2 in spleen T lymphocytes in the mice in model group was decreased significantly（P〈0.001）;Compared with model group,the contents of IL^-2 in spleen T lymphocytes in the mice in low,middle and high dose oviductus ranae groups were significantly increased（P〈0.001）;Spray drying was used to prepare microspheres,the optimum preparation was A3B1C1D1,the material ratio was 1∶3,the inlet temperature was 130℃,the outlet temperature was 100℃,and the feed flow rate was 30 mL·min^-1.Conclusion Oviductus ranae has immune regulation,they can be prepared into microspheres,the technology is stable and suitable for industrial production.