约氏疟原虫感染斯氏按蚊与大劣按蚊前后中肠前酚氧化酶的变化  

Changes of prophenoloxidase in the midguts of Anopheles stephensi and Anopheles dirus before and after infection with Plasmodium yoelii

作　　者：邱宗文[1] 张锡林[1] 徐文岳[1] 王英[1] 

机构地区：[1]第三军医大学基础医学部病原生物学教研室,重庆400038

出　　处：《第三军医大学学报》2004年第6期490-493,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 2 0 0 2 37)~~

摘　　要：目的　比较分析约氏疟原虫感染斯氏按蚊与大劣按蚊前后中肠前酚氧化酶 (prophenoloxidase ,PPO)的分布及其变化 ,探讨中肠PPO与按蚊抗疟原虫感染免疫防御反应的关系。方法　解剖分离约氏疟原虫感染前和感染后 3、5、7、11和 15d斯氏按蚊与大劣按蚊的中肠。利用烟草天蛾PPO抗体分别进行免疫组化染色和免疫印迹分析。结果　斯氏按蚊与大劣按蚊在未感染约氏疟原虫前的中肠循环管道内均已存在PPO ,而感染后循环管道内的PPO扩散到中肠组织间隙并聚集成片状、块状或条索状 ,大劣按蚊中肠PPO的聚集程度大于斯氏按蚊 ;在感染前和感染后不同时期免疫印迹均可检测到斯氏按蚊与大劣按蚊中肠PPO蛋白带 ,分子量约为 67× 10 3,感染后的PPO蛋白带型比感染前明显增强 ,而且大劣按蚊中肠PPO蛋白带型比斯氏按蚊尤为明显。结论　约氏疟原虫感染前按蚊中肠循环管道内存在的PPO可能是经与血淋巴相通的循环管道扩散而来 ,感染后导致的中肠内PPO不同分布及其变化可能与疟原虫感染按蚊密切相关 。Objective To investigate the relationship between the immune defence reaction against Plasmodium infection and the prophenoloxidase (PPO) of the midgut by comparative analysis of the distributions and the changes of PPO in the midgut of Anopheles stephensi and Anopheles dirus before and after infection with Plasmodium yoelii . Methods Midguts were dissected out from both Anopheles stephensi and Anopheles dirus at 3, 5, 7, 11 and 15 d before and after infection with Plasmodium yoelii . Immunohistochemistry and Western blotting were performed respectively on the collected midguts using Manduca Sexta PPO IgG polyantibody. Results PPO in the midguts from both Anopheles stephensi and Anopheles dirus was mainly located in the circulation conduit of midgut before infection with Plasmodium yoelii , but aggregated and distributed at the interspace of midguts as pieced or/and stripped forms after infection. Furthermore, PPO in the midgut of Anopheles dirus was more concentrated than that of Anopheles stephensi . Western blotting revealed that the PPO band with about molecular weight of 67×10 3 was detected in the midguts of both Anopheles stephensi and Anopheles dirus before and after Plasmodium yoelii infection. There was significant difference before and after infection, and the PPO band was obviously enhanced after infection. PPO bands in the midgut of Anopheles dirus were more prominent than those of Anopheles stephensi . Conclusion PPO in the midgut of Anopheles mosquitoes may come from the hemolymph by the circulation conduit before Plasmodium yoelii infection. However, the different distributions and changes of PPO in the midguts resulted from the Anopheles mosquitoes infected with Plasmodia may be closely correlated with Plasmodia infection, which may be of important physiological significance and may be involved in the immune defensive reaction against Plasmodium .

关 键 词：约氏疟原虫 斯氏按蚊 大劣按蚊 中肠 前酚氧化酶原 

分 类 号：Q554[生物学—生物化学] R382.31[医药卫生—医学寄生虫学]