抗蝰蛇毒IgY灌胃对蝰蛇伤小鼠保护作用的研究

Protection Effect of anti-viper venom IgY by intragastric administration against viper envenomation in mice

目的探讨抗蝰蛇蛇毒鸡卵黄抗体(immunoglobulin yolk,IgY)经腹腔注射或灌胃对蝰蛇伤小鼠保护作用,为其口服制剂的应用奠定理论基础。方法用蝰蛇原毒单一抗原免疫母鸡,收集第12天以后的鸡蛋,用水稀释法提取抗蝰蛇毒IgY;间接法ELISA检测IgY的效价和灌胃小鼠体内IgY的吸收时间;以胃排空率实验确定蛇伤前灌胃IgY的最佳时间,以腹腔注射和不同浓度IgY灌胃实验分别检验IgY对蝰蛇伤小鼠的保护作用。结果初次免疫后第12天开始收集抗体,水稀释法提取的IgY效价为1∶6400;不同浓度IgY灌胃后,2.5～3.5h胃排空率均达61.8%以上,血浆中抗体效价亦相应达高峰;腹腔注射和IgY灌胃实验对蛇伤小鼠的保护作用明显,小鼠的存活时间与阴性IgY组小鼠相比差异显著(P<0.05);同等效应下,灌胃IgY的有效剂量大约为腹腔注射的10～15倍。结论抗蝰蛇蛇毒IgY经腹腔注射和灌胃实验均能有效保护蝰蛇伤小鼠。

Objective The protection effect of anti-viper snake venom IgY was evaluated against viper venom by intragastric administration in mice, supplying the theory foundation for oral dosage forms of anti-viper snake venom IgY. Methods Single viper venom injected into the leghorn hen. Anti-viper snake venom IgY was extracted by water dilution. The tilter of IgY in vitro and the plasma concentration was observed in indirect ELISA assay after anti-viper snake venom IgY taken orally. The gastric emptying function test was used for determining optimization time after gastric administration of anti-viper snake venom IgY. The protection effect of anti-viper venom IgY was compared between intragastric administration and intraperitoneal injection in mice with viper envenomation. Results Anti-viper sake venom IgY was extracted by water dilution 12 days after the first immunization. The titer of anti-viper snake venom IgY was 1 ： 6400. In three concentrations（75.150.300 mg · 0. 5 ml^-1 · 20 g^-1 body weight）, at the time of 2. 5 hour to 3.5 hour after anti-viper snake venom IgY taken orally was excelled, and the gastric evacuation rate of mice was over 61.8%, with the plasm concentration in peak. The survival time of mice with viper snake envenomation was extremely prolonged （P=0. 000 〈0. 001）,after anti-viper snake venom IgY taken by intragastric administration or intraperitoheal injection. With similar effects of anti-viper snake venom IgY, the effective dose of intragastric administration was 10 to 15 times of that intraperitoneal injection. Conclusion Through two ways（by intragastric administration and by intraperitoneal injeetion）, anti-viper snake yenom IgY could effectively protect mice against viper snake venom.