Abstract：
OBJECTIVE: Human β-defensin 1 (hBD-1) is a antimicrobial peptide that is constantly secreted by oral tissues. Hangeshashinto (HST), a traditional Japanese medicine, has been reported to be effective against stomatitis. This study aimed to clarify the profile of HST by comparing the system of production of interleukin-1α (IL-1α) and hBD-1 in human oral mucosal epithelial cells with dexamethasone (DEX), a steroid used for the treatment of stomatitis.
METHODS: Human oral keratinocytes (HOK) were treated with HST, DEX, or HST components (baicalein, baicalin, berberine, and glycyrrhizin) for 24 h, and subsequently cultured for 24 h with or without Pam3CSK4 or lipopolysaccharide (LPS). The cell supernatants, total RNA, and intracellular proteins were collected, and changes in IL-1α and hBD-1 protein production and gene expression were evaluated using ELISA and RT-PCR. The phosphorylation of NF-kB and the cell proliferative ability of HOK were evaluated by western blotting and XTT assay, respectively.
RESULTS: DEX (0.01-10 μM) significantly suppressed IL-1α and hBD-1 production induced by either Pam3CSK4 or LPS, and also decreased cell growth. In contrast, HST inhibited Pam3CSK4- and LPS-induced IL-1α production at a concentration range of 12.5-100 μg/mL without affecting the cell proliferative capacity and hBD-1 production of HOK. Baicalein and baicalin, which are flavonoid ingredients of HST, showed anti-IL-1α production.
CONCLUSIONS: HST may be useful as a therapeutic agent for stomatitis and other inflammatory diseases of the oral cavity.
METHODS: Human oral keratinocytes (HOK) were treated with HST, DEX, or HST components (baicalein, baicalin, berberine, and glycyrrhizin) for 24 h, and subsequently cultured for 24 h with or without Pam3CSK4 or lipopolysaccharide (LPS). The cell supernatants, total RNA, and intracellular proteins were collected, and changes in IL-1α and hBD-1 protein production and gene expression were evaluated using ELISA and RT-PCR. The phosphorylation of NF-kB and the cell proliferative ability of HOK were evaluated by western blotting and XTT assay, respectively.
RESULTS: DEX (0.01-10 μM) significantly suppressed IL-1α and hBD-1 production induced by either Pam3CSK4 or LPS, and also decreased cell growth. In contrast, HST inhibited Pam3CSK4- and LPS-induced IL-1α production at a concentration range of 12.5-100 μg/mL without affecting the cell proliferative capacity and hBD-1 production of HOK. Baicalein and baicalin, which are flavonoid ingredients of HST, showed anti-IL-1α production.
CONCLUSIONS: HST may be useful as a therapeutic agent for stomatitis and other inflammatory diseases of the oral cavity.
摘要：
目的：人β-防御素1（hBD-1）是口腔组织不断分泌的抗菌肽。Hangeshashinto(HST),一种传统的日本药物，据报道对口腔炎有效。本研究旨在通过比较人口腔粘膜上皮细胞中白细胞介素-1α（IL-1α）和hBD-1与地塞米松（DEX）的产生系统来阐明HST的概况。用于治疗口腔炎的类固醇。
方法：用HST处理人口腔角质形成细胞（HOK），DEX,或HST成分(黄芩素，黄芩苷,小檗碱,和甘草酸)24小时，然后在有或没有Pam3CSK4或脂多糖（LPS）的情况下培养24小时。细胞上清液,总RNA，收集细胞内蛋白质，使用ELISA和RT-PCR评估IL-1α和hBD-1蛋白产生和基因表达的变化。通过蛋白质印迹和XTT实验评估NF-kB的磷酸化和HOK的细胞增殖能力。分别。
结果：DEX（0.01-10μM）显着抑制Pam3CSK4或LPS诱导的IL-1α和hBD-1的产生，也降低了细胞生长。相比之下,HST在12.5-100μg/mL的浓度范围内抑制Pam3CSK4-和LPS诱导的IL-1α的产生，而不影响细胞增殖能力和HOK的hBD-1产生。黄芩素和黄芩苷,是HST的类黄酮成分，显示抗IL-1α产生。
结论：HST可用作口腔炎和其他口腔炎性疾病的治疗剂。
方法：用HST处理人口腔角质形成细胞（HOK），DEX,或HST成分(黄芩素，黄芩苷,小檗碱,和甘草酸)24小时，然后在有或没有Pam3CSK4或脂多糖（LPS）的情况下培养24小时。细胞上清液,总RNA，收集细胞内蛋白质，使用ELISA和RT-PCR评估IL-1α和hBD-1蛋白产生和基因表达的变化。通过蛋白质印迹和XTT实验评估NF-kB的磷酸化和HOK的细胞增殖能力。分别。
结果：DEX（0.01-10μM）显着抑制Pam3CSK4或LPS诱导的IL-1α和hBD-1的产生，也降低了细胞生长。相比之下,HST在12.5-100μg/mL的浓度范围内抑制Pam3CSK4-和LPS诱导的IL-1α的产生，而不影响细胞增殖能力和HOK的hBD-1产生。黄芩素和黄芩苷,是HST的类黄酮成分，显示抗IL-1α产生。
结论：HST可用作口腔炎和其他口腔炎性疾病的治疗剂。
