目的探讨大劣按蚊血淋巴中丝氨酸蛋白酶与约氏疟原虫感染的相关性。方法首先利用二维电泳分离感染约氏疟原虫的和吸食正常血的大劣按蚊血淋巴蛋白;然后进行Western blot,用丝氨酸蛋白酶抗体进行识别,并比较感染组与正常对照组间的区别;选择感染后不同时相点对大劣按蚊蚊胃和唾液腺进腺镜检,以观察蚊体内疟原虫情况。结果Western blot显示,正常对照组无阳性蛋白点,感染组有2个阳性蛋白点,分子质量单位分别为44ku和27ku,等电点分别为8.0和5.0;镜检显示,感染7d时可见卵囊颗粒样病变,11d时卵囊发育不同步且黑化明显,之后检出的卵囊数逐渐减少。蚊唾液腺内未见子孢子。结论大劣按蚊血淋巴中的丝氨酸蛋白酶与约氏疟原虫感染相关,可能参与蚊抗疟原虫感染的黑化包被反应。
Objective To investigate the association between serine protease in the hemolymph of Anopheles dirus and infection with Plasmodium yoelii. Methods First, hemolymph proteins of An. dirus infected with P. yoelii and An. dirus fed on normal blood were separated by two dimensional polyacrylamide gel electrophoresis (2-DE). Then, Western blotting was carried out to identify serine proteases in hemolympb. The results of the two groups were compared. Finally, development of P. yoelii in the midgut and salivary glands of An. dirus was observed microscopically at different times. Results There ult in the control group, but two positive protein spots were found in the in fected group. Molecular weights of the two positive proteins were 44 ku and 27 ku, respectively. Isoelectric points (pI) of these proteins were 8.0 and 5.0, respectively. Microscopy results showed granulation at 7 d and different levels of development of oocysts and significant melanotic oocysts at 11 d. The number of oocysts declined over time. No sporozoites were found in mosquito salivary glands. Conclusion Serine protease in the hemolymph of An. dirus is associated with infection with P. yoelii and may play a role in its melanotic encapsulation.