日本血吸虫血红素加氧酶活性及免疫组化研究

Studies on the Activity and Immunohistochemistry of Heme Oxygenase in Schistosoma japonicum

目的 探讨日本血吸虫成虫是否具有血红素加氧酶 (HO)活性 ,并对其进行免疫学定位。方法 从日本血吸虫成虫匀浆中提取微粒体蛋白 ,与HO的特异性底物氯化血红素共孵育 ,观察血红素的降解情况及 pH值对降解活性的影响。制作日本血吸虫成虫组织切片和培养细胞涂片 ,用间接免疫荧光法及碱性磷酸酶免疫细胞化学试验研究HO在血吸虫体内的分布。结果 成虫的微粒体蛋白可在体外将血红素降解为胆红素 ,酶活性为 5 6 7nmol/(mg·min) ,最适 pH为 8 7。成虫组织切片显示 ,特异性荧光在虫体内呈散在性分布 ,细胞碱性磷酸酶免疫染色试验表明该酶位于成虫的细胞质。结论 首次证实日本血吸虫具有HO。

Objective To explore the activity of heme oxygenase and immunolocate the enzyme in the adult worms of Schistosoma japonicum .Methods Microsomal protein was isolated from the homogenate of adult S\^japonicum , heme degradation and effect of different pH conditions and buffers on degrading reaction were investigated by incubating microsomal protein with hemin. The slices of whole worm and cells of S\^japonicum were prepared, distribution of HO in schistosome was studied by immunofluorescent and alkaline phosphatase(AP) \|immunocytochemical assays.Results Microsomal protein of adult worms can degrade the heme in vitro , the activity being 56\^7 nmol bilirubin/(mg·min).The optimal pH was 8\^7. Immunofluorescent and AP\|immunocytochemical assays revealed that the HO distributed dispersively in the worm, and located in cytoplasm.Conclusion The presence of HO was firstly proved in S\^japonicum .