植物乳杆菌NDC 75017对Caco-2细胞中il-6表达的影响

Lactobacillus plantarum NDC 75017 affects il-6 gene expression in Caco-2 cells

摘要:

摘要:【目的】本研究通过体外实验，分析植物乳杆菌NDC75017 对免疫相关基因白细胞介素6(il-6)表达的影响，并进一步揭示其机制。【方法】植物乳杆菌NDC 75017作用于Caco-2细胞0、2、4、6、8、10和12 h，采用Real Time PCR方法检测il-6基因和toll样受体2(tlr2)的表达。植物乳杆菌NDC 75017与Caco-2细胞共培养0、0.5、1、2和4 h，用western blot方法检测NF-κB的磷酸化水平;NF-κB的特异性抑制剂PDTC预处理Caco-2细胞30 min后，再加入植物乳杆菌NDC 75017作用2 h，用Real Time PCR方法检测il-6基因及tlr2的表达量。【结果】植物乳杆菌NDC 75017诱导Caco-2细胞中il-6和tlr2基因的表达，并且分别在诱导8 h和6 h时表达量达到最大。植物乳杆菌NDC 75017 能够快速诱导NF-κB的磷酸化作用，在加入其特异性抑制剂PDTC后，il-6和tlr2基因的表达显著下降。【结论】植物乳杆菌NDC 75017能通过tlr2介导的NF-κB信号通路来诱导细胞因子il6短暂性的表达。

Abstract:

Abstract:［Objective］We tested the expression of immune-related gene interleukin 6 (il-6) in vitro to understand the influence from Lactobacillus plantarum NDC 75017 on host cells and further to reveal the regulatory mechanism．［Methods］Caco-2 cells were cocultured with L． plantarum NDC 75017 for 0，2，4，6，8，10 and 12 h，the total RNA were extracted; then the expressions of il-6 and tlr2 genes were analyzed by Real Time RT-PCR． The phosphorylation level of NF-κB was analyzed by Western Blot after the Caco-2 cells stimulation with L． plantarum NDC 75017 at 0，0. 5，1，2 and 4 h． Caco-2 cells were pretreated with pyrrolidine dithiocarbamate for 30 min before being treated with L． plantarum NDC 75017 for 2 h，then the total RNA was extracted and the expressions of il-6 and tlr2 genes were analyzed by Real Time RT-PCR． ［Results］Lactobacillus plantarum NDC 75017 could induce the expressions of il-6 and tlr2 in Caco-2 cells，the il-6 and tlr2 expressions peaked at 8 h and 6 h after cocultured with L．plantarum NDC 75017．L．plantarum NDC 75017 could rapidly activate the phosphorylation of NF-κB，and the expressions of il-6 and tlr2 were decreased notably after pretreated with pyrrolidine dithiocarbamate． ［Conclusion］L．plantarum NDC 75017 could up-regulate and then down-regulate the expression of il6 through rapidly activating tlr2-mediated NF-κB signaling pathway in Caco-2 cells．