抗尖吻蝮蛇毒卵黄抗体口服给药保护剂的研究

Research on Protectant for Immunoglobulin Yolk against the Venom of Deinagkistrodon acutus

【目的】比较硫糖铝和5种糖类对抗尖吻蝮蛇(Deinagkistrodon acutus)毒卵黄抗体(IgY)的保护作用,为开发抗尖吻蝮蛇毒IgY口服制剂提供依据。【方法】通过免疫白来航鸡(Gallus domesticus),从所产鸡蛋中制备抗尖吻蝮蛇毒IgY。在不同pH值(2.0,3.0)条件下,于IgY中加入相同剂量的保护剂,用ELISA法测定IgY活性;在pH值为2.0且加入条件下,加入等于或高于胃内正常质量浓度的胃蛋白酶,后加入不同剂量的硫糖铝,再用ELISA法测定IgY抗体活性;用不同剂量的保护剂与IgY混合后灌胃小鼠(Mus musculus),在不同时间点(1.5,2.5,3.5h)取血,采用ELISA和Western法测定血清中的IgY活性。【结果】在pH值为2.0和3.0条件下,硫糖铝对IgY抗体活性的保护效果均为最佳;在pH值为2.0且加入不小于胃内正常的胃蛋白酶质量浓度的条件下,体积分数在30%以上的硫糖铝均可提高IgY活性;IgY灌胃后的小鼠血清中,体积分数为50%的硫糖铝可有效保护IgY活性。【结论】加入硫糖铝作保护剂后能有效提高IgY对胃酸和胃蛋白酶的抵御作用。

[Purposes]This study compared the protective effect of sucralfate and five kinds of sugars（glucose,sucrose,synanthrin,trehalose,lactose）on activities of Anti-Deinagkistrodon acutus venom IgY in order to build a base for developing the oral IgY.[Methods]Antibodies against D.acutus venom came from eggs immunized with the snake venom which were laid by white leghorn hens（Gallus domesticus）.Under different pH values（2.0,3.0）,all protectants at the same concentrations were added into IgY solutions,and then activities of IgY in the mixture solutions were determined by ELISA.Under the same pH（2.0）,Pepsin in the normal or over-normal concentrations in mouse（Mus musculus）stomach and the sucralfate in various concentrations were added into IgY solutions,finally the activities of IgY in the mixture solutions were determined by ELISA.Using the different concentrations of sulcralfate to mix with IgY solution,then we gave mice the mixture solutions by oral administration,and at the different time points（1.5,2.5,3.5h）we took the blood of the mice and measured the activities of IgY in the serum using ELISA and Western blot.[Findings]The results showed that under pH 2.0and 3.0,the protection effects of sucralfate were higher than all of the other sugar protectants.Under pH2.0and the normal or over-normal concentrations of pepsin,more than 30%（V/V）concentrations of sucralfate may effectively protected IgY＇s activities.Following oral administration,50%（V/V）sucralfate concentration may effectively protected IgY＇s activities in mice serum.[Conclusions]Addition of sucralfate as a protective agent can effectively improve the resistance of IgY to gastric acid and pepsin.