鲤鱼过敏原(胶原蛋白)的体外模拟消化

The Vitro Simulated Digestion of Carp Allergens Collagen

首先采用酸法提取纯化鲤鱼过敏原（胶原蛋白）,依据美国药典提供的模拟胃液、肠液配方,分别利用胃蛋白酶、胰蛋白酶、胰凝乳蛋白酶对纯化的胶原蛋白进行酶解消化,以SDS-PAGE和免疫印迹分析测定纯化的胶原蛋白在体外模拟胃液、肠液中的消化稳定性及免疫原性.结果表明,鲤鱼胶原蛋白经模拟胃液（胃蛋白酶）消化15min出现较为明显的降解,消化1h的产物仍具有一定的免疫原性;经模拟肠液（胰蛋白酶）消化30min出现较为明显的降解,消化3h的产物仍具有一定的免疫原性;经模拟肠液（胰凝乳蛋白酶）消化1～4h均未出现明显降解,其消化产物仍具有较强的免疫原性.结果提示,鲤鱼胶原蛋白的耐消化性及免疫原性可能是导致食物性过敏反应的重要原因.

Firstly,acid method was used to extract the collagen from the skin and muscle of carp. Then pepsin,trypsin and chymotrypsin were used to digest the extracted collagen at different conditions based on the simulated gastric fluid＇s and intestinal fluid＇s formula provided by US pharmacopoeia. And the digested products were detected the digestion stability and immunogenicity in the simulated gastric fluid and intestinal fluid by SDS-PAGE and Western blot. It was found that the collagen could be digested by pepsin at 15 min, and it still had a certain degree of immunogenicity after 1 h. The collagen could be digested by trypsin at 30 min,and it still had a certain degree of immunogenicity after 3 h. The collagen wasn＇t almost digested by chymotrypsin at 1 ～ 4 h,and it still had the immunogenicity after 4 h. These results suggested that the digestion stability and immunogenicity of carp collagen were the major cause to induce the food allergy.