Liu Yan, Yang Shenghui, Xiao Jianhua, Yu Liang, Chen Li, Zou Ju, Wang Kegeng, Tan Sijie, Yu Zhengyang, Zeng Qingren
Department of Parasitology, School of Medicine, University of South China Hengyang, P.R. China ; Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study Hengyang, P.R. China.
Department of Preventive Medicine and Pathogenic Biology, School of Medicine, Hunan University of Chinese Medicine Changsha, P.R. China.
Int J Clin Exp Pathol. 2015 Feb 1;8(2):1247-58. eCollection 2015.
The present study was to determine the targeting effect of M13 phage peptide ZL4 (MppZL4) on Schistosoma japonicum (S.j). Mice infected with S.j were injected with MppZL4. Real-time PCR was used to detect the distribution and metabolism of MppZL4 in the livers and lungs of mice. In vivo refusion test was performed to detect the targeting of MppZL4. Western blotting was employed to determine the expression of MppZL4. Live imaging was used to detect the distribution of oligopeptide MppZL4. Immunohistochemistry was employed to determine MppZL4 location on adult S.j body surface. Gomori method was employed to detect the influence of oligopeptide MppZL4 on alkaline phosphatase activity. The distribution and metabolism of MppZL4 and M13KE are not significantly different from each other at each time point. The abundance of MppZL4 is changed as S.j migrates in mice. The targeted binding effect of MppZL4 varies at different stages. ZL4 oligopeptide targets S.j in mice. The specific binding sites of MppZL4 on S.j body are mainly located in syncytial cells. The binding sites of MppZL4 on S.j body surface might be ALP or ALP-related proteins. MppZL4 had targeted binding effect on S.j with its binding site being associated with proteins related to S.j alkaline phosphatase. S.j tegument had a specifically binding site with exogenous peptides, offering new means to explore the interactions between hosts and parasites. Additionally, MppZL4 can possibly be used as targeting molecules in worm-resistant drugs or as tracing molecules in imaging diagnosis technologies.
本研究旨在确定M13噬菌体肽ZL4(MppZL4)对日本血吸虫(S.j)的靶向作用。给感染日本血吸虫的小鼠注射MppZL4。采用实时荧光定量PCR检测MppZL4在小鼠肝脏和肺中的分布及代谢情况。进行体内再灌注试验以检测MppZL4的靶向性。采用蛋白质免疫印迹法测定MppZL4的表达。利用活体成像技术检测寡肽MppZL4的分布。采用免疫组织化学法确定MppZL4在日本血吸虫成虫体表的定位。采用Gomori法检测寡肽MppZL4对碱性磷酸酶活性的影响。在各个时间点,MppZL4和M13KE的分布及代谢情况无显著差异。随着日本血吸虫在小鼠体内迁移,MppZL4的丰度发生变化。MppZL4的靶向结合作用在不同阶段有所不同。ZL4寡肽可靶向小鼠体内的日本血吸虫。MppZL4在日本血吸虫虫体上的特异性结合位点主要位于合体细胞。MppZL4在日本血吸虫体表的结合位点可能是碱性磷酸酶(ALP)或与ALP相关的蛋白。MppZL4对日本血吸虫具有靶向结合作用,其结合位点与日本血吸虫碱性磷酸酶相关蛋白有关。日本血吸虫体表存在与外源性肽的特异性结合位点,为探索宿主与寄生虫之间的相互作用提供了新手段。此外,MppZL4有可能用作抗蠕虫药物的靶向分子或成像诊断技术中的示踪分子。