Effect of piperazine (diethylenediamine) on the moulting, proteorne expression and pyrophosphatase activity of Ascaris suum lung-stage larvae
Effect of piperazine (diethylenediamine) on the moulting, proteorne expression and pyrophosphatase activity of Ascaris suum lung-stage larvae
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DOI：
10.1016/j.actatropica.2006.08.007
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发表时间：
2006-10-01
期刊：
影响因子：
2.7
通讯作者：
Tsuji, Naotoshi
中科院分区：
文献类型：
--
作者：
Islam, M. Khyrul;Miyoshi, Takeharu;Tsuji, Naotoshi
二乙撑二胺（diethylenediamine）是一种广泛用于防治动物和鸟类蛔虫的驱虫药。在这项研究中，我们表明，哌嗪块处理猪蛔虫幼虫蜕皮和发育过程中，影响幼虫蛋白质组表达谱。A.从感染的兔的肺获得的猪肺-L3期（LL 3）在存在递增浓度的硫酸哌嗪（Pzes）的RPMI培养基中培养。结果表明，Pzes对A. suum LL 3以剂量依赖性的方式，并且在50 mM浓度下蜕皮被完全阻断（100%）。然后我们研究了A的变化。猪LL 3蛋白质组在Pzes暴露后的表达模式，使用二维（21）电泳。Pzes暴露抑制了在从蜕皮幼虫制备的2D凝胶上分辨的200多个可见蛋白质斑点中的至少16个主要蛋白质斑点在未蜕皮LU中的表达（即，L4期）。Pzes暴露还抑制未蜕皮的LL 3中的13个免疫原性蛋白质点的表达。更重要的是，Pzes暴露抑制了蜕皮特异性酶的活性，无机焦磷酸酶A。猪天冬氨酸蛋白酶（AsPPase），提高了26%。通过免疫印迹和免疫荧光染色检测到，Pzes暴露后天然AsPPase的表达也减少。透射电子显微镜观察表明，Pzes干扰角质层的生长和蜕皮，并造成肠道组织的损伤。结果表明，A.猪LL 3可能成为筛选具有抗虫功能新型驱虫药的理想模型，而猪A.猪LL 3-Pzes蛋白可作为蛔虫蜕皮特异性蛋白的鉴定工具。(c)2006 Elsevier B. V.保留所有权利。
Piperazine (diethylenediamine) is an anthelmintic widely used against animal and bird ascariasis. In this study, we show that treatment with piperazine blocks Ascaris suum larval moulting and development processes and affects larval proteome expression profiles. A. suum lung-stage L3 (LL3) obtained from an infected rabbit's lungs were cultured in RPMI medium in the presence of increasing concentrations of piperazine sulfate (Pzes). Our results showed that Pzes potently inhibited moulting of A. suum LL3 in a dose-dependent manner and that moulting was completely blocked (100%) at 50 mM concentrations. We then examined the changes in A. suum LL3 proteome expression patterns following Pzes exposure using two-dimensional (21)) electrophoresis. Pzes exposure inhibited expression of at least 16 major protein spots in unmoulted LU out of more than 200 visible protein spots resolved on 2D gels prepared from moulted larvae (i.e., lung-stage L4). Pzes exposure also inhibited expression of 13 immunogenic protein spots in unmoulted LL3. More importantly, Pzes exposure inhibited activity of a moulting-specific enzyme, inorganic pyrophosphatase of A. suum (AsPPase), by 26%. Expression of native AsPPase was also reduced following Pzes exposure as detected by immunoblotting and immunofluorescent staining. Transmission electron microscopy showed that Pzes interfered with growth and ecdysis of the cuticle and caused damage to gut tissues of the larvae. Our results suggest that A. suum LL3 may become a suitable model to screening new-class anthelmintics with antimoulting functions and that A. suum LL3-Pzes may serve as a useful tool for identification of moulting-specific potential proteins in Ascaris roundworms. (c) 2006 Elsevier B.V. All rights reserved.