文章摘要
徐栋花,孙可一,季晓辉.乳酸杆菌对LPS诱导的THP-1细胞炎症性细胞因子释放的调节作用[J].南京医科大学学报,2011,(7):962~969
乳酸杆菌对LPS诱导的THP-1细胞炎症性细胞因子释放的调节作用
Regulatory effects of lactic acid bacteria on the secretion of LPS-induced inflammatory cytokines in THP-1 cells
投稿时间:2011-01-24  
DOI:10.7655
中文关键词: 乳酸杆菌  THP-1细胞  炎性细胞因子  TNF-α  NF-κB
英文关键词: lactic acid bacteria  THP-1 cell  inflammatory cytokines  TNF-α  NF-κB
基金项目:江苏省现代病原学重点实验室开放课题
作者单位
徐栋花 南京医科大学微生物学与免疫学系,江苏 南京 210029 
孙可一 南京医科大学微生物学与免疫学系,江苏 南京 210029 
季晓辉 南京医科大学微生物学与免疫学系,江苏 南京 210029 
摘要点击次数: 2124
全文下载次数: 169
中文摘要:
      目的:探讨副干酪乳酸杆菌(Lactobacillus paracasei,L.para)与嗜酸性乳酸杆菌(Lactobacillus acidophilus,L.acid)对脂多糖(LPS)诱导THP-1细胞分泌肿瘤坏死因子(TNF)-α?白细胞介素(IL)-12 p40?转化生长因子(TGF)-β的调节作用?方法:先将THP-1细胞在佛波酯(PMA)的刺激作用下活化?分化为巨噬细胞样细胞,再进行LPS诱生炎性细胞因子实验,实验分为空白对照组?LPS处理组?单独L.para处理组?L.para与LPS共处理组?单独L.acid处理组及L.acid与LPS共处理组?以RT-PCR法检测THP-1细胞TNF-α mRNA水平的表达;以ELISA法检测细胞培养上清中TNF-α?IL-12 p40和TGF-β水平的变化;以Western blot方法检测胞内未磷酸化及磷酸化IκB-α蛋白水平的表达;细胞免疫荧光技术观察NF-κB p65的亚细胞定位,TransAM核蛋白定量分析法检测THP-1细胞核内NF-κB(p65/p50)的水平?结果:① LPS能显著刺激PMA诱导分化的THP-1细胞分泌炎性细胞因子TNF-α?IL-12 p40和TGF-β;乳酸杆菌L.para及L.acid单独作用时也能诱导细胞因子TNF-α和IL-12 p40的产生,但L.para诱导的水平较低,而L.acid诱导的水平与LPS相似,两种乳酸杆菌对TGF-β的诱导作用则均显著高于LPS;而当两菌分别与LPS共同作用于PMA诱导分化的THP-1细胞时,都能显著抑制LPS刺激细胞诱生的TNF-α和IL-12 p40,并以L.para的抑制作用更明显,但两种细菌均对TGF-β没有抑制作用,反而进一步促进其分泌?②与结果①相平行,LPS?L.para及L.acid单独作用时,均能使PMA诱导分化的THP-1细胞胞核内磷酸化IκB-α的水平增高?非磷酸化IκB-α水平降低,核内NF-κB(p65/p50)的水平增高,但L.para的作用相对较弱;而当L.para或L.acid与LPS共同作用时则显著抑制LPS刺激所诱导的IκB-α的磷酸化及NF-κB的核转位作用?结论:乳酸杆菌L.para?L.acid分别与LPS共同作用时,均能通过抑制LPS刺激细胞所诱生的炎性细胞因子TNF-α?IL-12 p40的水平而发挥炎症调节作用,这一抑制作用的机制涉及抑制 NF-κB信号通路?而且,乳酸杆菌的这一作用在不同菌种之间存在明显差异?
英文摘要:
      Objective:To explore the regulatory effects of lactic acid bacteria on the secretion of tumor necrosis factor(TNF)-α, interleukin(IL)-12 p40 and transforming growth factor(TGF)-β in THP-1 cells induced by lipopolysaccharide(LPS). Methods: THP-1 cells were firstly stimulated by phorbol 12-myristate 13-acetate(PMA) to differentiate into macrophage-like cells, and were secondly treated by LPS to induce inflammatory cytokines. Six groups were designed, including blank control group, LPS-treated group, L.para-treated alone group, L.para plus LPS-treated group, L.acid-treated alone group and L.acid plus LPS-treated group. The levels of TNF-α mRNA in THP-1 cells were determined by RT-PCR. TNF-α, IL-12 p40 and TGF-β in the supernatant of THP-1 cells were assayed by ELISA. The subcellular localization of NF-κB p65 was observed by immunofluorescence, while the quantitation of NF-κB(p65/p50) in THP-1 nucleus was detected by TransAMTM NF-κB kit. Results:① The secretions of TNF-α, IL-12 p40 and TGF-β in PMA-induced macrophage-like THP-1 cells were remarkably increased when treated by LPS, the same as L.para or L.acid treated alone respectively. However, TNF-α and IL-12 p40 in L.para-treated alone group were lower than those in LPS-treated group, whereas TNF-α and IL-12 p40 in L.acid-treated alone group were similar to the LPS-treated group. TGF-β induced by L.para or L.acid was strikingly higher than that of LPS. Interestingly, TNF-α and IL-12 p40 in PMA-induced macrophage-like THP-1 cells stimulated by LPS were notably inhibited when cells were treated with L.para plus LPS or L.acid plus LPS. TNF-α and IL-12 p40 in L.para plus LPS-treated group were much lower than in LPS-treated group. No inhibition of TGF-β were observed in PMA-induced macrophage-like THP-1 cells when treated with both lactobacilli.② Similar to result one, phosphorylation of IκB-α and nuclear NF-κB(p65/p50) in PMA-induced macrophage-like THP-1 cells increased in LPS-treated group, L.para-treated alone group and L.acid-treated alone group, while non-phosphorylation of IκB-α accordingly decreased. The regulatory effect of L.para was a little weaker than L.acid. Phosphorylation of IκB-α and nuclear translocation of NF-κB were significantly reduced in PMA-induced macrophage-like THP-1 cells when treated with L.para plus LPS or L.acid plus LPS. Conclusion:L.para and L.acid can inhibit the secretion of LPS-induced inflammatory cytokines TNF-α and IL-12 p40 when they interacted with LPS respectively. The negative regulation of lactic acid bacteria was related to inhibition of NF-κB signaling. Besides, the regulatory effects of lactic acid bacteria were evidently different among species.
查看全文   查看/发表评论  下载PDF阅读器