摘要
目的观察日本血吸虫成虫单酚氧化酶(monophenoloxidase,MPO)和二酚氧化酶(diphenoloxidase,DPO)的同工酶酶谱形式。方法将收集到的42d活成虫置含0.05%戊巴比妥钠的RPMI1640培养基中,23℃孵育8h,分别研磨雌、雄成虫呈匀浆,显色单酚氧化酶的样品匀浆时另加5%的TX-100。超声粉碎、低温高速离心取上清(含PO),进行聚丙烯酰胺凝胶电泳(PAGE)及酶染色后分析两种酚氧化酶同工酶酶谱形式。结果单酚氧化酶的同工酶酶谱与二酚氧化酶的同工酶酶谱基本一致,电泳位置相同,表现为迁移率相同的一条带,但是单酚氧化酶的酶谱谱带较二酚氧化酶的酶谱显色弱。同时,雌虫及雄虫的单酚氧化酶和二酚氧化酶亦表现为迁移率相同的一条主带;但雄虫的酶带显色反应较雌虫弱。结论日本血吸虫成虫体内酚氧化酶存在两种形式:单酚氧化酶和二酚氧化酶。而且不仅雌性虫含有酚氧化酶,雄性成虫也含有少量的酚氧化酶且两者酶分子相同。雄性成虫含有少量酚氧化酶的生理意义有待进一步研究。
Objective To study the isoenzyme zymogram of monophenol oxidase (MPO) and diphenol oxidase(DPO) of Schistosoma japonicum adult worm. Methods Living adult worms(aged 42d) were incubated in RPMI1640 containing 0.05% sodium phenobarbital for 8h at 23℃. Then female worms and male worms were separated and collected. The worms were homogenized and fragmented by ultra sonication. The suspensions were centrifuged and the supernatants were collected as the protein samples. The isoenzyme zymogram of phenol oxidase (MPO; DPO) was analyzed by means of polyacrylamide gel electrophoresis (PAGE). Results The isoenzyme zymogram patterns of DPO and MPO were almost identical except the color intensity of MPO was less than that of DPO. The similarity of isoenzyme zymogram patterns from females and males were also found, including a major band and a same migrating rate, but the color intensity of the zymogram of males was less than that of females. Conclusions There were two kinds of phenol oxidase forms(MPO; DPO) in Schistosoma japonicum. Their isoenzyme zymogram patterns were similar. Furthermore the phenol oxidase of S.japonicum does exist not only in female worms but also in male worms. Their zymogram patterns were similar in the number and migrating rate of enzyme band, but the color intensity of the male zymogram pattern was lower. The understanding of the physiological role of PO in male worms needs further investigation.
出处
《热带医学杂志》
CAS
2003年第3期298-300,共3页
Journal of Tropical Medicine
基金
国家自然科学基金资助项目(No.30240074)
湖北省教育厅资助项目(No.2001A14009)。
关键词
日本血吸虫
单酚氧化酶
二酚氧化酶
酶谱
Schistosoma japonicum
monophenol oxidase
diphenol oxidase
zymogram
