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目的探讨日本血吸虫成虫呕吐物和排泄分泌物的蛋白质组成,寻找高丰度蛋白分子。方法将日本血吸虫成虫浸泡于无菌水中15～30min,收集成虫的呕吐和排泄分泌物。通过蛋白质二维电泳,考马斯亮蓝染色与飞行质谱分析,确定各蛋白点的基因种类。结果日本血吸虫成虫的呕吐及排泄分泌物的二维电泳获得1012个蛋白斑点,456个斑点的蛋白质种类被质谱分析成功鉴定。这些蛋白可归为139种,其中76种为虫源性蛋白,主要是由一些与血吸虫生命代谢、生长发育、免疫调控相关的蛋白组成,其中烯醇化酶、70kDa热休克蛋白(HSP70)、果糖二磷酸醛缩酶、三磷酸甘油醛脱氢酶、磷酸甘油酸激酶、谷胱甘肽转移酶、抑免蛋白、硫氧还蛋白过氧化物酶、14-3-3蛋白等含量丰富,是高丰度的排泄分泌蛋白。结论本研究揭示了日本血吸虫成虫呕吐和排泄分泌物的蛋白组成,为进一步开发用于控制血吸虫病或其他寄生虫病的疫苗、新的治疗药物及诊断方法创造了条件。
Objective To investigate the protein composition of adult vomitus and excretory secretion of Schistosoma japonicum and to search for high abundance protein molecules. Methods Adult Schistosoma japonicum soaked in sterile water for 15 ~ 30min to collect the vomit and excretion of adult worms. By protein two-dimensional electrophoresis, Coomassie brilliant blue staining and flight mass spectrometry to determine the gene types of each protein spots. Results 1012 protein spots were obtained by two-dimensional electrophoresis of the secretions of Schistosoma japonicum adults with vomiting and excretion. The protein types of 456 spots were successfully identified by mass spectrometry. These proteins can be grouped into 139 species, of which 76 are insect-derived proteins, which are mainly composed of proteins related to the metabolism, growth and immune regulation of Schistosoma japonicum. Enolase, 70 kDa heat shock protein (HSP70), fructose Diphosphate aldolase, glyceraldehyde triphosphate dehydrogenase, phosphoglycerate kinase, glutathione transferase, inhibin, thioredoxin peroxidase, 14-3-3 protein content is rich, is High abundance of secreted secreted protein. Conclusions The present study revealed the protein composition of adult vomit and excretion secretions of Schistosoma japonicum and provided the conditions for the further development of vaccines, new therapeutic drugs and diagnostic methods for controlling schistosomiasis or other parasitic diseases.