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目的观察旋毛虫幼虫侵入前后小鼠肠上皮细胞蛋白的变化,筛选幼虫侵入相关蛋白。方法将旋毛虫感染性幼虫接种至小鼠小肠上皮细胞(intestinal epithelial cells,IECs)单层,于37℃5%CO2条件下培养2h,提取细胞蛋白,进行SDS-PAGE与Western blot分析,并与正常IECs细胞蛋白比较。结果 SDS-PAGE分析表明,IECs与感染性幼虫共孵育后的IECs蛋白带数为44条,与正常IECs对照组的相同;Western blot显示,正常IECs蛋白不能被旋毛虫感染鼠血清识别,与幼虫共孵育后的IECs蛋白有16个组分带能被旋毛虫感染鼠血清识别,分子质量单位分别为125、118、106、98、70、64、50、47、46、40、34、33、31、29、27、26ku。结论小鼠IECs与旋毛虫幼虫共培养可产生能被旋毛虫感染鼠血清识别的细胞蛋白,该组蛋白可能是旋毛虫侵入相关蛋白。
Objective To observe the changes of protein in intestinal epithelial cells of mice before and after invasion of Trichinella spiralis larvae and screen larvae for invasion of related proteins. Methods Infected larvae of Trichinella spiralis were inoculated into monolayers of mouse intestinal epithelial cells (IECs) and cultured for 2 h at 37 ℃ under 5% CO2. Cell proteins were extracted and analyzed by SDS-PAGE and Western blot. Comparison of normal IECs cell proteins. Results SDS-PAGE analysis showed that the number of IECs bands incubated with IECs and infectious larvae was 44, which was the same as that of the normal IECs control group. Western blot showed that the normal IECs protein could not be recognized by the serum of Trichinella infected mice, After co-incubation of IECs protein 16 components can be infected with Trichinella antibody serum identification, molecular mass units were 125,118,106,98,70,64,50,47,46,40,34,33, 31,29,27,26ku. Conclusions Incubation of mouse IECs with Trichinella spiralis larvae can produce cell proteins that can be recognized by the serum of Trichinella spiralis infected mice, which may be involved in invasion of Trichinella spiralis related proteins.