青蒿琥酯作用后日本血吸虫童虫蛋白质组学分析

Proteomic analysis of Schistosoma japonicum schistosomula after artesunate treatment

目的探讨青蒿琥酯处理前后日本血吸虫童虫蛋白质组的差异表达，研究青蒿琥酯抗日本血吸虫的分子机制。方法新西兰兔经贴片法感染日本血吸虫尾蚴（2000条／只）第7天一次性灌服青蒿琥酯（实验组，8mg／kg）和1％羧甲基纤维素钠（对照组），第10天经肝门静脉系统灌流收集童虫，观察虫体形态，提取总蛋白。应用同位素标记相对和绝对定量技术（iTRAQ技术）鉴定青蒿琥酯处理前后13本血吸虫童虫的差异蛋白质组，通过数据库检索和生物信息学分析差异表达的蛋白质。结果青蓠琥酯处理后日本血吸虫童虫与对照组相比，形态较为粗短细小，并存在明显的发育不良或畸形。iTRAQ技术检测显示，处理后的血吸虫童虫有75个（含3个未知蛋白）蛋白存在明显表达差异。其中表达量上调的蛋白23个，下调的蛋白52个。结论差异表达的蛋白主要参与了童虫的代谢、生物调节和细胞稳态等过程，同时与应激、细胞组织和生成等密切相关。

Six New Zealand rabbits were infected with 2 000 cercariae by using coating method and orally administered with once administration of Art at a dose of 8 mg/kg on the 7^th day after infection. The parallel control group was only administered with 1% sodium carboxymethylcellulose（n=3）. On the 10^th day, schistosomula were obtained from rabbits by perfusion and total protein of Schistosomula was extracted. The proteins from two groups were identified by isobaric tags for relative and absolute quantitation （iTRAQ） techniques. Bioinformatics analysis of differentially expressed proteins was made and their functions were confirmed by database searching. Results Compared with control group, Schistosoma japonicum schistosomula were more stubby after Art treatment, and there was obrious dysplasia and deformity, iTRAQ analysis showed that there were 75 proteins （including 3 uncharacterized proteins） differentially expressed after Art treatment, among which 23 were up-regulated proteins and 52 were down-regulated proteins. Conclusions The differentially expressed proteins are mainly involved in metabolic processes, regulation of biological process and cellular homeostasis. Moreover, those proteins also play an important role in stress response, cell organization and biogenesis.