University of Münster, Institute of Pharmaceutical Biology and Phytochemistry, Münster, Germany.
Umeå University, Medical Biochemistry and Biophysics, Umeå, Sweden.
PLoS One. 2014 Jan 9;9(1):e84836. doi: 10.1371/journal.pone.0084836. eCollection 2014.
Traditional Asian and African medicine use immature okra fruits (Abelmoschus esculentus) as mucilaginous food to combat gastritis. Its effectiveness is due to polysaccharides that inhibit the adhesion of Helicobacter pylori to stomach tissue. The present study investigates the antiadhesive effect in mechanistic detail.
A standardized aqueous fresh extract (Okra FE) from immature okra fruits was used for a quantitative in vitro adhesion assay with FITC-labled H. pylori J99, 2 clinical isolates, AGS cells, and fluorescence-activated cell sorting. Bacterial adhesins affected by FE were pinpointed using a dot-blot overlay assay with immobilized Lewis(b), sialyl-Lewis(a), H-1, laminin, and fibronectin. (125)I-radiolabeled Okra FE polymer served for binding studies to different H. pylori strains and interaction experiments with BabA and SabA. Iron nanoparticles with different coatings were used to investigate the influence of the charge-dependence of an interaction on the H. pylori surface.
Okra FE dose-dependently (0.2 to 2 mg/mL) inhibited H. pylori binding to AGS cells. FE inhibited the adhesive binding of membrane proteins BabA, SabA, and HpA to its specific ligands. Radiolabeled compounds from FE bound non-specifically to different strains of H. pylori, as well as to BabA/SabA deficient mutants, indicating an interaction with a still-unknown membrane structure in the vicinity of the adhesins. The binding depended on the charge of the inhibitors. Okra FE did not lead to subsequent feedback regulation or increased expression of adhesins or virulence factors.
Non-specific interactions between high molecular compounds from okra fruits and the H. pylori surface lead to strong antiadhesive effects.
传统的亚洲和非洲医学使用未成熟的秋葵果实(Abelmoschus esculentus)作为粘液食物来治疗胃炎。其有效性归因于多糖,它可以抑制幽门螺杆菌与胃组织的粘附。本研究从机制上详细研究了这种抗粘附作用。
使用未成熟秋葵果实的标准化水性新鲜提取物(Okra FE)进行定量体外粘附测定,使用 FITC 标记的 H. pylori J99、2 株临床分离株、AGS 细胞和荧光激活细胞分选。使用固定化 Lewis(b)、唾液酸化 Lewis(a)、H-1、层粘连蛋白和纤维连接蛋白的斑点印迹覆盖测定法,确定 FE 影响的细菌粘附素。(125)I 放射性标记的 Okra FE 聚合物用于与不同的 H. pylori 菌株结合研究,并与 BabA 和 SabA 进行相互作用实验。使用具有不同涂层的铁纳米粒子来研究相互作用的电荷依赖性对 H. pylori 表面的影响。
秋葵 FE 呈剂量依赖性(0.2 至 2 mg/mL)抑制 H. pylori 与 AGS 细胞的结合。FE 抑制膜蛋白 BabA、SabA 和 HpA 与其特定配体的粘附结合。FE 中的放射性标记化合物非特异性地结合到不同的 H. pylori 菌株以及 BabA/SabA 缺失突变体,表明与粘附素附近的未知膜结构发生相互作用。这种结合取决于抑制剂的电荷。秋葵 FE 不会导致随后的反馈调节或粘附素或毒力因子的表达增加。
来自秋葵果实的高分子化合物与 H. pylori 表面之间的非特异性相互作用导致强烈的抗粘附作用。