共生细菌来源的脂多糖在水稻中作为巨噬细胞激活物质的功能表征。

Functional characterization of lipopolysaccharide derived from symbiotic bacteria in rice as a macrophage-activating substance.

机构信息

Department of Faculty of Medicine, Kagawa University, 1750-1 Ikenobe, Miki-cho, Kida-gun, Kagawa-ken, 761-0793, Japan.

出版信息

Anticancer Res. 2011 Jul;31(7):2467-76.

Abstract

BACKGROUND AND AIM

Lipopolysaccharide derived from a symbiotic bacterium in wheat (Pantoea agglomerans, LPSp) has shown multiple positive effects, such as prophylactic, antiallergic and antitumour effects, without serious side-effects. LPSp has differential biological activities in comparison to other LPS, such as those from Escherichia coli (LPSe). The only difference between LPSp and LPSe is in the O-antigen polysaccharide structure (O-PS). This led us to the hypothesis that the O-PS structure would seem to participate in biological activities. Thus, the characterization of properties of O-PS in LPS is of the utmost importance for understanding cell activation in the maintenance of homeostasis. However, little is known about the correlation between the O-PS structure of LPS and its biological activities. In this study, we extracted LPS derived from a symbiotic bacterium in rice (strain A46, related species of Pantoea), which has a long history of use in foods, and investigated its putative structures and functions.

MATERIALS AND METHODS

LPS derived from strain A46 was prepared using a hot phenol extraction method. The properties of LPS-A46 were analysed by thin-layer chromatography, Tricine SDS-PAGE and Western blotting. The function of LPS-A46 was analyzed by quantative real-time PCR and flow cytometry using THP-1 cells and Peripheral blood mononuclear cell (PBMC) derived macrophages.

RESULTS

In Tricine SDS-PAGE, high molecular mass LPS-A46 had a molecular mass lower than that of LPSp. In Western blotting, LPS-A46 reacted with lipid A antibody but did not react with an O-PS antibody of LPSp. In comparison to other LPS, LPS-A46 induced a differential cytokine gene expression profile in THP-1 cells and PBMC-derived macrophages.

CONCLUSION

The present study suggests that LPS derived from symbiotic bacterium in rice is a bioactive functional LPS which may have different functional activities compared to other types of LPS.

摘要

背景与目的

来源于小麦共生菌（成团泛菌，LPSp）的脂多糖具有多种积极作用，如预防、抗过敏和抗肿瘤作用，且没有严重的副作用。与其他脂多糖（如大肠杆菌脂多糖，LPSe）相比，LPSp 具有不同的生物学活性。LPSp 与 LPSe 的唯一区别在于 O-抗原多糖结构（O-PS）。这使我们假设 O-PS 结构似乎参与了生物学活性。因此，LPS 中 O-PS 结构的特性对于理解细胞激活在维持体内平衡中的作用至关重要。然而，人们对 LPS 的 O-PS 结构与其生物学活性之间的相关性知之甚少。在这项研究中，我们提取了来源于水稻共生菌（菌株 A46，泛菌的相关种）的 LPS，该共生菌在食品中有悠久的应用历史，并研究了其可能的结构和功能。

材料与方法

采用热酚提取法提取 A46 菌株 LPS。采用薄层层析、Tricine SDS-PAGE 和 Western blot 分析 LPS-A46 的性质。通过定量实时 PCR 和流式细胞术分析 LPS-A46 在 THP-1 细胞和外周血单核细胞（PBMC）衍生的巨噬细胞中的功能。

结果

在 Tricine SDS-PAGE 中，高分子质量 LPS-A46 的分子量低于 LPSp。在 Western blot 中，LPS-A46 与脂多糖 A 抗体反应，但与 LPSp 的 O-PS 抗体不反应。与其他 LPS 相比，LPS-A46 在 THP-1 细胞和 PBMC 衍生的巨噬细胞中诱导了不同的细胞因子基因表达谱。