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-KB 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-kappaB, 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-kappaB signaling pathway in Caco-2 cells.
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-KB 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-kappaB, 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-kappaB signaling pathway in Caco-2 cells.