约氏疟原虫感染诱导大劣按蚊AdSP1和AdSP3的转录  

Enhanced transcript abundance of AdsP1 and AdsP3 from Anopheles dirus haemocytes after Plasmodium yoelii infection

作　　者：徐文岳[1] 黄复生[1] 夏丽莎[1] 段建华[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486-489,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　观察丝氨酸蛋白酶AdSP1和AdSP3在大劣按蚊主要组织中的表达 ,分析血餐和约氏疟原虫感染对大劣按蚊血细胞AdSP1和AdSP3转录的影响。方法　离心收集血细胞 ,解剖分离大劣按蚊蚊胃和唾液腺 ,提取相应的总RNA ,RT PCR扩增 ;然后将同批 3～ 5d龄大劣按蚊分为正常组 (N)、吸正常血组 (B)和感染约氏疟原虫组 (I) ,分别在血餐后 1、2、3、4、7d和 11d ,收集 3组各 5 0只雌大劣按蚊血细胞和提取总RNA ,以Ads7为内参照 ,进行半定量RT PCR分析。结果　丝氨酸蛋白酶AdSP1和AdSP3在大劣按蚊血细胞和唾液腺中表达 ,不在蚊胃表达 ;吸血和约氏疟原虫感染后 ,大劣按蚊血细胞中AdSP1和AdSP3的转录上调 ,尤以约氏疟原虫黑化期间明显。结论　AdSP1和AdSP3可能参与大劣按蚊对约氏疟原虫的黑化反应 ,并可能是大劣按蚊的PPAE。Objective To investigate the expression patterns of serine proteases Adsp1 and Adsp3 in the main tissue of Anopheles dirus and the effects of blood feeding and Plasmodium infection on the transcript abundance of Adsp1 and Adsp3 from Anopheles dirus haemocytes. Methods Haemocytes were collected by centrifugation. The midguts and salivary glands were dissected from 50 adult female Anopheles dirus of 3-5 d old for the extraction of total RNA for amplification by RT PCR. Anopheles dirus of 3-5 d old were divided into normal group (N), blood feeding group (B), and Plasmodium yoelii infection group (I). Haemocytes of 50 adult female Anopheles dirus from each group after blood feeding were also collected, and the total RNA was isolated at 1, 2, 3, 4, 7, and 11 d, respectively. Then, the same volume (10 μl) of total RNA was analyzed by semi quantitative RT PCR with Ads7 as the internal control. Agarose gel analysis was performed on PCR products from each group. Results Expression of both Adsp1 and Adsp3 mRNA was found in the haemocytes and salivary glands, but not in the midguts. Semi quantitative RT PCR indicated that transcript abundance of AdsP1 and AdsP3 from Anopheles dirus haemocytes was enhanced after blood feeding and Plasmodium yoelii infection, especially during melanotic encapsulation of Plasmodium yoelii . Conclusion AdsP1 and AdsP3 may be involved in the melanotic encapsulation response of Plasmodium yoelii by Anopheles dirus , and may be the prophenoloxidase activating enzyme of Anopheles dirus .

关 键 词：大劣按蚁 约氏疟原虫 丝氨酸蛋白酶 RT-PCR 

分 类 号：R382.31[医药卫生—医学寄生虫学] R384.1[医药卫生—基础医学]