大肠杆菌诱导的促炎细胞因子的产生受 MAP 激酶和 G 蛋白调节，而不受 Akt 调节：与进化群和耐药模式的关系。

Escherichia coli-induced productions of pro-inflammatory cytokines are regulated by MAP kinases and G-protein but not by Akt: Relationship with phylogenetic groups and resistance patterns.

机构信息

Université de Nantes, EA 3826 thérapeutiques cliniques et expérimentales des infections, UFR Médecine, 44000 Nantes, France.

出版信息

Cytokine. 2011 Nov;56(2):290-7. doi: 10.1016/j.cyto.2011.06.015. Epub 2011 Jul 18.

DOI:10.1016/j.cyto.2011.06.015

PMID:21764597

Abstract

INTRODUCTION

We investigated the role of PI3-K, MAP kinases, and heterotrimeric G proteins in inducing cytokines production in human whole blood cultures stimulated by viable Escherichia coli (E. coli) clinical strains.

MATERIALS AND METHODS

We used eight E. coli strains that belong to different phylogenetic groups and presented by different antibiotic resistance patterns. Whole blood from healthy volunteers was incubated at 37°C for 150min, with lipopolysaccharide (LPS) from E. coli O111:B4 or selected viable E. coli clinical strains, with or without SB202190 (p38 inhibitor), PD98059 (ERK inhibitor), PTX (pertussis toxin; heterotrimeric G proteins inhibitor), wortmaninn (PI3-K inhibitor). The TNF-α, IL-1β, IL-10 and IFN-γ concentrations were measured in culture supernatants (ELISA).

RESULTS

IL-10 and IFN-γ were not detectable. Susceptible strains induced higher TNF-α and IL-1β productions than β-lactam resistant strains (p<0.05), with no difference between phylogenetic groups. A transformed strain carrying a plasmid-mediated AmpC-β-lactamase gene (CMY-2) induced lower TNF-α and IL-1β production than the parent wild type strain (p<0.05). SB202190 (p38 inhibitor) and PD98059 (ERK inhibitor) reduced TNF-α concentrations by, respectively, 80% (p<0.05) and 50% (p<0.05). Wortmaninn (PI3-K inhibitor) had no significant effect. PTX (heterotrimeric G proteins inhibitor) altered TNF-α production after viable bacteria stimulation (1.7-fold increase; p<0.05) but not after LPS (TLR-4) stimulation. Regarding IL-1β, wortmaninn, SB202190 and PTX had no significant effect whereas PD98059 significantly decreased production in whole cell cultures (p<0.05).

CONCLUSION

Susceptible strains induce greater TNF-α and IL-1β productions than resistant strains. ERK kinase plays a major role in viable E. coli strains inducing TNF-α and IL-1β production. E. coli exerts an effect on the pertussis toxin-sensitive G-protein through a TLR-4-independent mechanism.

摘要

简介

我们研究了 PI3-K、MAP 激酶和异三聚体 G 蛋白在诱导人全血培养物中细胞因子产生中的作用，这些培养物由活的大肠杆菌（E. coli）临床株刺激。

材料与方法

我们使用了 8 株属于不同进化群且具有不同抗生素耐药模式的大肠杆菌菌株。来自健康志愿者的全血在 37°C 下孵育 150min，用大肠杆菌 O111:B4 的脂多糖（LPS）或选定的活的大肠杆菌临床株，或不加 SB202190（p38 抑制剂）、PD98059（ERK 抑制剂）、PTX（百日咳毒素；异三聚体 G 蛋白抑制剂）、wortmaninn（PI3-K 抑制剂）刺激。通过 ELISA 测量培养上清液中的 TNF-α、IL-1β、IL-10 和 IFN-γ 浓度。

结果

未检测到 IL-10 和 IFN-γ。敏感株诱导的 TNF-α 和 IL-1β 产量高于β-内酰胺耐药株（p<0.05），但进化群之间无差异。携带质粒介导的 AmpC-β-内酰胺酶基因（CMY-2）的转化株诱导的 TNF-α 和 IL-1β 产量低于亲本野生型株（p<0.05）。SB202190（p38 抑制剂）和 PD98059（ERK 抑制剂）分别使 TNF-α 浓度降低 80%（p<0.05）和 50%（p<0.05）。wortmaninn（PI3-K 抑制剂）无显著影响。PTX（异三聚体 G 蛋白抑制剂）改变了活菌刺激后的 TNF-α 产生（增加 1.7 倍；p<0.05），但不改变 LPS（TLR-4）刺激后的 TNF-α 产生。关于 IL-1β，wortmaninn、SB202190 和 PTX 无显著影响，而 PD98059 显著降低全细胞培养物中的产量（p<0.05）。