日本血吸虫不同发育期虫卵内抗氧化酶检测的研究

摘要

The aim is to examine antioxidant levels in Schistosoma japonicum (S. japonicum) eggs in different develop ment stage, and evaluate the function of antioxidant staining to identify eggs trapped in human tissue. Mice were randomly di vided into four groups: 50 or 90 day post infection groups, 30 or 90 day post treatment groups, with viable S. japonicum in the first two groups and no viable S. japonicum in the last two groups, respectively. Antioxidase stained eggs was separated from the liver and colonic mucous of different groups with 3,3 Dimethylbenzidine and hydrogen peroxide as its substrates, and then the enzyme levels of different groups were comparatively observed by coloration and distribution for evaluation. Compared with undyed eggs, the positive dark brownish yellow was the most visible when the free eggs were staining at room temperature and oscillation for 30 min. No coloration was observed once the eggs boiled with DAB substrates for 30 min. The positive rates in these four groups were 92. 34% , 46. 03% , 13. 68% and 0. 00 %. The positive degree showed that immature eggs presented dark brownish yellow, while mature eggs were deep yellow located in miracidium body. The current degradation eggs were partly dyed in which most immature eggs and the degradation eggs and shell showed no positive reaction. The findings suggest that the viable S. japonicum eggs are rich in antioxidase, particularly in immature eggs, while the enzyme levels fade away as eggs develop from maturity to death. So the results suggest that antioxidase staining of S. japonicum eggs can be a markerrnto discriminate viable or dead eggs.%目的 检测日本血吸虫(Sj)不同发育期虫卵内抗氧化酶水平,分析其是否可作为鉴别虫卵死与活的标志物.方法 采用3,3-二甲基联苯胺盐酸盐(DAB)+H2O2底物作为抗氧化酶检测方法对完整Sj虫卵进行染色观察,并探索其染色条件.建立Sj感染后50 d和90 d的小鼠模型和吡喹酮治疗后30 d和90 d的小鼠模型.用最佳染色条件对从各模型组小鼠肝肠组织分出的虫卵作检测,比较观察对未成熟卵、成熟卵、近期变性卵和远期变性卵的显色程度与其分布来评价抗氧化酶水平,分析其用于判别Sj虫卵死与活的可能性.结果 与不做染色虫卵比,DAB加入完整游离活卵中经室温震动反应30 min时,所呈深棕黄色阳性反应的反差最明显;DAB对沸腾处理30 min虫卵染色,未见呈显色反应;DAB对Sj感染50 d、90 d及治后30 d和90 d的模型鼠肝肠来源虫卵染色的阳性率分别为92.34%、46.03%、13.68%和0.00 %.其阳性反应程度:未成熟虫卵普遍呈深棕黄色;成熟卵多呈深黄色,分布于毛蚴体部;近期变性卵中部分有显色;远期变性卵和卵壳均无见显色.结论 Sj活虫卵内均富含抗氧化酶成分,其水平则以未成熟卵内为最高,且随着虫卵发育成熟至死亡而逐渐减低.提示Sj虫卵作抗氧化酶染色可作为虫卵死与活鉴别的标志物.