| 肖勇健,刘卓然,赵飞骏,余坚,曾铁兵.梅毒螺旋体纤连蛋白结合蛋白Tp0136前炎症活性的研究.[J].中南医学科学杂志.,2016,(3):263-266, 295. |
| 梅毒螺旋体纤连蛋白结合蛋白Tp0136前炎症活性的研究 |
| Proinflammatory Activity of Recombinant Fibronectin-BindingProteins Tp0136 of Treponema Pallidum |
| 投稿时间：2015-11-17 修订日期：2016-04-20 |
| DOI： |
| 中文关键词: 梅毒螺旋体 纤连蛋白结合蛋白 Tp0136 前炎症细胞因子 核因子kappa B |
| 英文关键词:Treponema pallidum fibronectin-binding protein (FnBP) Tp0136 proinflammatory cytokines NF-κB |
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| 中文摘要: |
| 目的 初步研究梅毒螺旋体纤连蛋白结合蛋白Tp0136潜在的前炎症活性。方法构建重组质粒pET-28a/Tp0136并诱导其表达目的蛋白；将不同浓度(1、2、5、10 μg/mL)的rTp0136分别刺激THP-1源性巨噬细胞,ELISA法分别检测其产生前炎症细胞因子IL-1β、IL-6和TNF-α的水平；并同时用二硫代氨基甲酸吡咯烷(PDTC)预先处理巨噬细胞,检测相应细胞因子分泌量。结果成功构建了pET-28a/Tp0136重组质粒,并诱导表达和纯化出rTp0136；重组蛋白能诱导巨噬细胞产生IL-1β、IL-6和TNF-α,并且呈浓度和时间依赖性；重组蛋白刺激经PDTC预处理后的巨噬细胞,其IL-1β、IL-6和TNF-α的分泌水平分别降至32%、35%和27%。结论rTp0136可诱导巨噬细胞产生前炎症细胞因子,该过程可能受NF-κB调控。 |
| 英文摘要: |
| Objective To explore potential proinflammatory activity of the Treponema pallidum fibronectin-binding protein (FnBP) Tp0136.MethodsThe recombinant pET-28a/Tp0136 was constructed and induced to express the targeting protein in E.coli BL21.In vitro various concentrations of protein was used to stimulate human macrophages derived from THP-1 cell line at different time.ELISA was performed to detect the level of proinflammatory cytokines IL-1β,IL-6 and TNF-α;Recombinant protein was used to stimulate the macrophages pre-treated with PDTC,and ELISA was performed to detect the releasing IL-1β,IL-6 and TNF-α respectively.ResultsThe recombinant plasmid was constructed and the corresponding recombinant protein was obtained successfully;rTp0136 could induce production of IL-1β,IL-6 and TNF-α in dose- and time-dependent manners significantly.Secretion of IL-1β,IL-6 and TNF-α decreased to 32%,35% and 27% respectively when macrophages was pre-treated with PDTC.ConclusionThe data suggested that the FnBP Tp0136 could activate the THP-derived human macrophages to release the proinflammatory cytokines,which might be regulated via NF-κB. |
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