Department of Orthodontics, Nihon University School of Dentistry, 1-8-13 Kanda Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
Division of Clinical Research, Dental Research Center, Nihon University School of Dentistry, 1-8-13 Kanda Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
Int J Med Sci. 2017 Sep 19;14(12):1173-1180. doi: 10.7150/ijms.20387. eCollection 2017.
Electrolytically-generated acid functional water (FW) is obtained by electrolyzing low concentrations of saline. Although it has been widely used in clinical practice with various purposes, the underlying mechanisms of action involved have not been fully elucidated so far. We used the human cervical cancer-derived fibroblastic cell line (HeLa), to examine the cytokine secretion profile following FW treatment in the present study. FW stimulation significantly induced the secretion of basic fibroblast growth factor (bFGF) and extracellular matrix metalloproteinase inducer (EMMPRIN). The effect of both factors on osteoblast-like MC3T3-E1 cells was further examined by stimulating the cells with the conditioned medium of FW-stimulated HeLa cells. However, the conditioned medium failed to induce IL-6 secretion. The MC3T3-E1 cells were further stimulated with recombinant bFGF or EMMPRIN or a combination of both factors. Intriguingly, bFGF-stimulated IL-6 induction was totally inhibited by EMMPRIN. Pretreatment with the specific inhibitor of nuclear factor-kappa B (NF-κB) drastically inhibited IL-6 secretion indicating that bFGF-induced IL-6 expression was dependent on NF-κB activation. The phosphorylation status of NF-κB p65 subunit was further examined. The results indicated that EMMPRIN inhibited bFGF-induced NF-κB p65 phosphorylation. These findings suggest that bFGF can induce IL-6 secretion in MC3T3-E1 cells through NF-κB activation. As EMMPRIN inhibited bFGF-induced IL-6 secretion by reducing the p65 subunit phosphorylation, it might be concluded that bFGF and EMMPRIN crosstalk in their respective signaling pathways.
电解生成的酸性功能水（FW）是通过电解低浓度的盐水获得的。尽管它已广泛应用于临床实践中，具有多种用途，但迄今为止，其作用机制尚未完全阐明。本研究使用人宫颈癌细胞衍生的成纤维细胞系（HeLa），研究 FW 处理后细胞因子分泌谱的变化。FW 刺激显著诱导碱性成纤维细胞生长因子（bFGF）和细胞外基质金属蛋白酶诱导因子（EMMPRIN）的分泌。通过用 FW 刺激的 HeLa 细胞的条件培养基刺激成骨样 MC3T3-E1 细胞，进一步研究了这两种因子对细胞的影响。然而，该条件培养基未能诱导 IL-6 的分泌。进一步用重组 bFGF 或 EMMPRIN 或两者的组合刺激 MC3T3-E1 细胞。有趣的是，bFGF 刺激的 IL-6 诱导被 EMMPRIN 完全抑制。用核因子-kappa B（NF-κB）的特异性抑制剂预处理可明显抑制 IL-6 的分泌，表明 bFGF 诱导的 IL-6 表达依赖于 NF-κB 的激活。进一步检测 NF-κB p65 亚基的磷酸化状态。结果表明，EMMPRIN 抑制 bFGF 诱导的 NF-κB p65 磷酸化。这些发现表明，bFGF 可以通过 NF-κB 激活诱导 MC3T3-E1 细胞中 IL-6 的分泌。由于 EMMPRIN 通过减少 p65 亚基磷酸化抑制 bFGF 诱导的 IL-6 分泌，因此可以得出结论，bFGF 和 EMMPRIN 在其各自的信号通路中相互作用。
