Department of Periodontology and Endodontology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan.
J Periodontal Res. 2010 Feb;45(1):79-86. doi: 10.1111/j.1600-0765.2009.01203.x. Epub 2009 Jul 8.
Oral epithelial cells help to prevent against bacterial infection in the oral cavity by producing antimicrobial peptides (AMPs). A broad-spectrum AMP, calprotectin (a complex of S100A8 and S100A9 proteins), is expressed by oral epithelial cells and is up-regulated by interleukin-1alpha (IL-1alpha). Shosaikoto (SST) is a traditional Japanese herbal medicine that has immunomodulatory effects and is reported to enhance the levels of IL-1alpha in epithelial cells. The purpose of this study was to investigate the effect of SST on the expression of calprotectin and other AMPs through the regulation of IL-1alpha in oral epithelial cells.
Human oral epithelial cells (TR146) were cultured with SST (at concentrations ranging from 10 to 250 microg/mL) in the presence or absence of anti-IL-1alpha or IL-1 receptor antagonist. The expression of S100A8- and S100A9-specific mRNAs was examined by northern blotting. Calprotectin expression and IL-1alpha secretion were investigated by immunofluorescent staining or ELISA. The expression of other AMPs and IL-1alpha was analyzed by RT-PCR and by quantitative real-time PCR.
Shosaikoto (25 microg/mL) significantly increased the expression of S100A8- and S100A9-specific mRNAs and calprotectin protein. Shosaikoto increased S100A7 expression, but had no effect on the expression of other AMPs. The expression of IL-1alpha-specific mRNA and its protein were slightly increased by SST. A neutralizing antibody against IL-1alpha or IL-1 receptor antagonist inhibited SST up-regulated S100A8/S100A9 mRNA expression.
These results suggest that SST increases the expression of calprotectin and S100A7 in oral epithelial cells. In response to SST, up-regulation of calprotectin may be partially induced via IL-1alpha.
口腔上皮细胞通过产生抗菌肽（AMPs）来帮助预防口腔内的细菌感染。 calprotectin（S100A8 和 S100A9 蛋白的复合物）是一种广谱 AMP，由口腔上皮细胞表达，并受白细胞介素-1α（IL-1α）的上调。 十全大补汤（SST）是一种具有免疫调节作用的传统日本草药，据报道可提高上皮细胞中 IL-1α 的水平。本研究旨在通过调节上皮细胞中的 IL-1α 来研究 SST 对 calprotectin 和其他 AMPs 表达的影响。
用 SST（浓度范围为 10 至 250μg/mL）在存在或不存在抗 IL-1α 或 IL-1 受体拮抗剂的情况下培养人口腔上皮细胞（TR146）。通过 northern 印迹法检查 S100A8 和 S100A9 特异性 mRNA 的表达。通过免疫荧光染色或 ELISA 研究 calprotectin 的表达和 IL-1α 的分泌。通过 RT-PCR 和定量实时 PCR 分析其他 AMPs 和 IL-1α 的表达。
十全大补汤（25μg/mL）显着增加了 S100A8 和 S100A9 特异性 mRNA 和 calprotectin 蛋白的表达。十全大补汤增加了 S100A7 的表达，但对其他 AMPs 的表达没有影响。 SST 轻度增加了 IL-1α 特异性 mRNA 的表达及其蛋白。抗 IL-1α 或 IL-1 受体拮抗剂的中和抗体抑制了 SST 上调的 S100A8/S100A9 mRNA 表达。
这些结果表明，SST 增加了口腔上皮细胞中 calprotectin 和 S100A7 的表达。响应 SST，calprotectin 的上调可能部分通过 IL-1α 诱导。
