中国寄生虫学与寄生虫病杂志 ›› 2004, Vol. 22 ›› Issue (3): 8-159.

• 论著 • 上一篇    下一篇

诱导型一氧化氮合酶在感染日本血吸虫小鼠肝组织中的表达

龙小纯1,李雍龙1*,Andreas Ruppel 2
  

  1. 1 华中科技大学同济医学院病原生物学系,武汉 430030 2 德国海德堡大学热带卫生研究所,德国海德堡
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2004-06-30 发布日期:2004-06-30

Expression of Inducible Nitric Oxide Synthase in the Livers of Mice Infected with Schistosoma japonicum

LONG Xiao-chun,LI Yong-long*,Andreas Ruppel
  

  1. Department of Parasitology,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,430030,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2004-06-30 Published:2004-06-30

摘要:   目的 研究诱导型一氧化氮合酶 (iNOS)在感染日本血吸虫小鼠肝脏中的动态表达。 方法 日本血吸虫尾蚴经皮肤感染NMRI小鼠,感染后第0、21、28、38和45天取其肝组织,用逆转录聚合酶链反应 (RTPCR)半定量法检测iNOS转录水平的动态表达 ,用蛋白质印迹法 (Westernblotting)测定表达的蛋白变化,并用免疫组化方法确定iNOS在肝组织中的表达定位。 结果 RTPCR结果显示,未感染小鼠肝组织中未见iNOS的表达 ,感染后第21天可见到iNOS在肝组织中的转录表达,第28天明显表达,第38天达峰值,第45天下降,与第38天相比,差异无显著性意义 (P>0.05)。Westernblotting结果表明,感染后第38天和第45天分别可见到iNOS的表达。间接荧光抗体试验结果表明,iNOS主要在肝虫卵肉芽肿细胞中表达。 结论 日本血吸虫能诱导宿主肝组织iNOS的动态表达,肝组织中的虫卵可能与iNOS表达有关。

关键词: 日本血吸虫, 诱导型一氧化氮合酶, 虫卵肉芽肿, 逆转录聚合酶链反应, 蛋白质印迹法

Abstract:  Objective To study the expression of inducible nitric oxide synthase (iNOS) in livers of mice infected with Schistosoma japonicum. Methods The livers of NMRI mice infected with S. japonicum were collected on day 21,28,38,45 post infection(p.i.), total RNA of livers were extracted and kinetics of the mRNA expression of iNOS were detected by RT-PCR, the protein expression of iNOS was then confirmed by Western blotting and the distribution of iNOS in the infected liver was determined by immunohistochemical methods. Results The mRNA expression of iNOS was not detectable in the uninfected liver, iNOS mRNA expression was detected on day 21 p.i, the expression increased on day 28 p.i and peaked on day 38 p.i, then decreased slightly on day 45 p.i. Western blotting showed an iNOS expression in the livers only on day 38, 45 p.i. IFA test showed that the expression of iNOS was maily distributed in the granuloma of the livers. Conclusion S. japonicum infection can induce the expression of iNOS in a time-dependent manner in the liver of the host,and eggs may be the main factor in inducing the expression.

Key words: Schistosoma japonicum, Inducible nitric oxide synthase, Egg granuloma, RT-PCR, Western blotting