中国农业科学 ›› 2012, Vol. 45 ›› Issue (8): 1621-1626.doi: 10.3864/j.issn.0578-1752.2012.08.018

• 兽医 • 上一篇    下一篇

鼠李糖乳杆菌对巨噬细胞先天性免疫应答的调节作用

 黄琴, 黄怡, 崔志文, 李雅丽, 李卫芬, 余东游   

  1. 1.浙江大学动物科学学院/教育部动物分子营养学重点实验室,杭州 310058
    2.广西大学动物科学技术学院,南宁 530005
  • 收稿日期:2011-08-15 出版日期:2012-04-15 发布日期:2012-02-10
  • 通讯作者: 通信作者李卫芬,Tel:0571-86986730;E-mail: wfli@zju.edu.cn
  • 作者简介:黄 琴,E-mail: huangqinqin83@163.com
  • 基金资助:

    浙江省重大科技专项(2006C12086)、国家重点基础研究发展计划(2009CB118705)、博士学科点专项科研基金(20110101110101)

Modulation of Lactobacillus rhamnosus on Innate Immune Responses in Macrophages

1.浙江大学动物科学学院/教育部动物分子营养学重点实验室,杭州 310058
2.广西大学动物科学技术学院,南宁 530005   

  1. 1.浙江大学动物科学学院/教育部动物分子营养学重点实验室,杭州 310058
    2.广西大学动物科学技术学院,南宁 530005
  • Received:2011-08-15 Online:2012-04-15 Published:2012-02-10

摘要: 【目的】通过测定先天性免疫效应因子分泌水平的变化,研究鼠李糖乳杆菌(Lactobacillus rhamnosus)体外对巨噬细胞先天性免疫应答调节的影响。【方法】将培养好的鼠巨噬细胞系RAW264.7细胞分为4组,分别用PBS(CT组,对照组)、Escherichia coli K88(EC组)和Lactobacillus rhamnosus(LR组)处理12 h,以及先用Lactobacillus rhamnosus预处理1 h,再用Escherichia coli K88处理12 h(LR-EC组)。试验结束时,收集各组的细胞培养液,用ELISA检测TNF-α、IFN-γ、IL-1β、IL-6、IL-12、IL-8、IL-10,以及PGE2和 的含量。【结果】结果表明,CT组(对照组)可产生TNF-α、IFN-γ、IL-8和IL-10,几乎不产生IL-1β、IL-6和IL-12。与对照组相比,EC组TNF-α、IFN-γ、IL-8、IL-10及PGE2和 含量均极显著提高(P<0.01),且可大量产生IL-1β、IL-6和IL-12;LR组TNF-α、IFN-γ、IL-10及PGE2和 也极显著提高(P<0.01),IL-8显著增加(P<0.05),同样不产生IL-1β和IL-6,但可产生微量的IL-12(P<0.01);LR-EC组所有的促炎细胞因子(TNF-α、IFN-γ、IL-1β、IL-6、IL-12)、趋化因子IL-8和PGE2均极显著高于EC组(P<0.01),但IL-10和 含量都显著低于EC组(P<0.01)。【结论】在体外试验条件下,鼠李糖乳杆菌作为益生菌对生理状态下巨噬细胞先天性免疫应答的刺激作用远低于病原菌,不会产生炎症反应,但可极大增强受感染巨噬细胞的促炎免疫应答水平,且可能具有避免过度炎症反应的作用。

Abstract: 【Objective】Lactobacillus rhamnosus (L. rhamnosas) was examined for its capacity to facilitate the innate immune responses of macrophage cell in vitro in terms of innate immune factor production. 【Method】 The murine macrophage cell line, RAW 264. 7, was treated with PBS (designated as group CT, control group), Escherichia coli K88(designated as group EC)and L. rhamnosus(designated as group LR)for 12 h,respectively, as well as treated with L. rhamnosus prior for 1 h, and then exposed to E. coli K88 for 12 h(designated as group LR-EC). After incubation, culture supernatants were collected for further analysis of TNF-α, IFN-γ, IL-1β, IL-6, IL-12, IL-8, IL-10, PGE2 and   using ELISA. 【Result】Results showed that TNF-α, IFN-γ, IL-8 and IL-10 production was observed in group CT (the control group),whereas the concentrations of IL-1β, IL-6 and IL-12 were below the detection limits. Production of TNF-α, IFN-γ, IL-8, IL-10, PGE2 and   in group EC increased significantly(P<0.01)compared to group CT, and higher concentrations of IL-1β, IL-6 and IL-12 were also detectd in group EC. Higher levels of TNF-α, IFN-γ, IL-10, PGE2,     (P<0.01)and IL-8(P<0.05)were determined in group LR while no detectable effect on IL-1β and IL-6 was found. Moreover,  the release of all tested proinflammatory cytokines(TNF-α, IFN-γ, IL-1β, IL-6, IL-12), chemotactic factor (IL-8) and PGE2 in  group LR-EC was up-regulated remarkably(P<0.01)while secretion of IL-10 and   was noticeably suppressed in comparison to group EC(P<0.01). 【Conclusion】 It was concluded that probiotics strain L. rhamnosus is capable of triggering a moderate innate immune response on direct contact with murine macrophage cell line RAW264.7. Furthermore, L. rhamnosus could enhance proinflammatory response to infected macrophages; meanwhile, it might prevent excessive inflammatory reaction in vitro.

Key words: RAW264.7 cell, Lactobacillus rhamnosus, Escherichia coli K88, cytokine, chemotactic factor, PGE2