Cytokine related plasminogen activator activity and regulation in human dental pulp cells

人牙髓细胞中细胞因子相关纤溶酶原激活剂的活性和调节

• 批准号: 17592001
• 负责人: HASHIZUME Hideki
• 金额: $ 2.24万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17592001/
• 关键词: cytokine; plasminogen avtivator; urokinase; NF-κB; IL-1β; TNF-α; uPA; チロシンのリン酸化

Plasminogen activator (PA) is the enzyme converting plasminogen to its active form, plasmin involved in various physiological and pathological phenomenon. The conversion is catalysed by two types of plasminogen activator (PA), urokinase-type PA and tissue-type PA (tPA). We investigated effect of the inflammatory cytokin IL-lβ and TNF-a on PA secretion in human dental pulp cells. When the cells were stimulated by IL-1β and TNF-a, PA activity in the medium was clearly increased in a time-and dose-dependent manner. The PA activity in the medium was reduced after immunoprecipitation using anti-uPA antibody, and uPA protein was detected in the immunoprecipitated fraction by Western blotting. Anti-tPA antibody failed to show such observations. In the IL-1β and TNF-a-stimulated cells, uPA mRNA expression was enhanced but less tPA mRNA. The IL-1β and TNF-a-stimulated uPA mRNA expresson and PA activities in cell lysate and medium were reduced by herbimycin A and genistein, tyrosin kinase inhibitors, and pyrolidinedithiocarbamate, an NFκB inhibitor, and were augmented by the tyrosine phosphatase inhibitor sodium orthovanadate. These observations suggest that IL-1β and TNF-a stimulates uPA production via NFκB and tyrosine phosphorylation activation and the enzyme secretion, and uPA/plasmin system appears to be involved in inflammation in human dental pulp.

纤溶酶原激活剂（PA）是将纤溶酶原转化为活性形式的酶，纤溶酶参与各种物理和病理现象。转化率由两种类型的纤溶酶原激活剂（PA），尿激酶型PA和组织型PA（TPA）催化。我们研究了炎症性细胞因子IL-Lβ和TNF-A对人牙纸浆细胞中PA分泌的影响。当通过IL-1β和TNF-A刺激细胞时，培养基中的PA活性以时间依赖性的方式明显增加。使用抗UPA抗体免疫沉淀后，培养基中的PA活性降低，并通过蛋白质印迹在免疫沉淀的部分中检测到UPA蛋白。抗TPA抗体未能显示出这种观察结果。在IL-1β和TNF-A刺激的细胞中，UPA mRNA表达增强，但TPA mRNA较少。在细胞裂解液和培养基中，IL-1β和TNF-A刺激的UPA mRNA表达和PA活性被草药霉素A和染料偶200S，酪氨酸激酶抑制剂和硫代硫代氨基酯，NFκBB抑制剂，酪氨酸抑制剂，酪氨酸抑制剂，并被酪氨酸磷酸磷酸酶抑制剂增强。这些观察结果表明，IL-1β和TNF-A通过NFκB和酪氨酸磷酸化激活和酶分泌刺激UPA产生，并且UPA/纤溶酶系统似乎参与了人类牙齿牙髓的炎症。