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广东血管圆线虫诱导的活性氧应激的半乳糖凝集素-10 的 mRNA 水平。

The mRNA level of the galectin-10 of Angiostrongylus cantonensis induced by reactive oxygen stress.

机构信息

Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, Guangdong, China.

出版信息

Parasitol Res. 2013 Mar;112(3):933-43. doi: 10.1007/s00436-012-3210-5. Epub 2012 Dec 9.

Abstract

Galectin plays an important role in host-parasite interactions. In this study, we identified a novel gene encoding galectin-10 (AcGal-10) from the cDNA library of Angiostrongylus cantonensis and characterized its biological role in the parasite. Sequence and phylogeny analysis showed that AcGal-10 is related to other galectin family members with the conserved loci (H(84)-D(86)-R(88)-V(96)-N(98)-W(105)-E(108)-R(110)). The mRNA level of AcGal-10 was expressed in reactive oxygen stress radicals. We have identified two proteins of A. cantonensis galectin-10 gene, one of which was reported (AcGAL10-W) and the others is AcGAL-10-M. In addition, recombinant AcGal-10 (rAcGal-10) was constructed into the pGEX-4T-1 plasmid, purified, and finally confirmed by SDS-PAGE and LC-MS. Hemagglutination assay showed that the minimum concentration of rAcGAL10-W and rAcGAL10-M required for the hemagglutination of BALB/c mice erythrocyte was 25 μg/mL, and the carbohydrate-binding ability showed no difference between rAcGAL10-W and rAcGAL10-M. The mRNA levels of AcGal-10 were indeed expressed higher after stimulation with H(2)O(2) and recombinant A. cantonensis galectin-10. A mutation of AcGal-10 was also found, but there was no significant difference compared with the wild type. Furthermore, we also confirmed that recombinant AcGal-10 plays a role in the activation of the microglia. In conclusion, the report here showed that AcGal-10 may be an important molecule related to infection of A. cantonensis.

摘要

半乳糖凝集素在宿主-寄生虫相互作用中发挥着重要作用。在这项研究中,我们从广州管圆线虫 cDNA 文库中鉴定出一种新的半乳糖凝集素-10(AcGal-10)基因,并对其在寄生虫中的生物学作用进行了表征。序列和系统发育分析表明,AcGal-10 与其他半乳糖家族成员具有保守的基因座(H(84)-D(86)-R(88)-V(96)-N(98)-W(105)-E(108)-R(110))。AcGal-10 的 mRNA 水平在活性氧应激自由基中表达。我们已经鉴定出两种 A. cantonensis 半乳糖凝集素-10 基因的蛋白质,其中一种已报道(AcGAL10-W),另一种是 AcGAL-10-M。此外,重组 AcGal-10(rAcGal-10)构建到 pGEX-4T-1 质粒中,经 SDS-PAGE 和 LC-MS 最终确认。血凝试验表明,rAcGAL10-W 和 rAcGAL10-M 使 BALB/c 小鼠红细胞发生凝集的最小浓度分别为 25μg/mL,并且 rAcGAL10-W 和 rAcGAL10-M 的糖结合能力没有差异。用 H(2)O(2)和重组 A. cantonensis 半乳糖凝集素-10 刺激后,AcGal-10 的 mRNA 水平确实表达更高。还发现 AcGal-10 发生了突变,但与野生型相比没有显著差异。此外,我们还证实重组 AcGal-10 在外周血单个核细胞激活小胶质细胞中发挥作用。综上所述,本报告表明 AcGal-10 可能是与 A. cantonensis 感染相关的重要分子。

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