Cellular functional character of extracellular matrix on dental pulpitis

牙髓炎细胞外基质的细胞功能特征

• 批准号: 12671862
• 负责人: MATSUSHIMA Kiyoshi
• 金额: $ 2.11万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671862/
• 关键词: Dental pulpitis; Dental pulp cell; Gingival cell; IL-6; TNF-alpha; Plasminogen activator; Tissue inhibitoe metalloprotease; Plasminogen Activator; Tissue inhibitor metaloprotease; plasminogen activator; plasmin; 炎症性サイトカイン; MMPs

The extracellular matrix (ECM) plays an important role in the growth and degeneration of cells As a result of infection and host immunoreaction, ECM is damaged by matrix metalloproteases (MMPs). Plasmin, which is produced from limited degradation of plasminogen by plasminogen activator (PA), activated latent MMPs and stimulates the synthesis of kinin. The inflammatory cytokines such as IL-1beta, IL-6 and TNF-alpha are synthesized by invasion of bacteria or virus. The effect of various inflammatory cytokines on the PA/plasmin system in human dental pulp cells was examined and compared with the effect on human gingival cells in order to characterize dental pulp cells in pulpitis. Dental pulp and gingival cells which were obtained from same donor were cultured in alpha-MEM with 10 % fetal calf serum and under 5 % CO_2 in air at 37 ℃. Both cells in a con fluent-stage were incubated for 24 h in medium containing 2 % PCS and treated with various cytokines (IL-1beta, IL-6 and TNF-alpha). The PA activity in conditioned medium was measured using fluorescent substrate. The expression of tPA mRNA was analyzed by RT-PCR. The PA activity of dental pulp cells was higher and the expression of tPA mRNA was stronger than that of gingival cells when both cells were incubated with IL-6 and TNF-alpha. The inflammatory cytokines (IL-1beta, IL-6 and TNF-alpha) stimulated PA activity via an enhancement of tPA gene expression and MMP-2 activity, and tPA activated by these cytokines stimulated MMP-9, Thus the inflammatory cytokines may be involved in extracellular matrix degradation through a stimulation of the PA/plasmin system of human dental pulp cells. Dental pulp cells were different from gingival cells in the response of the IL-6 and TNF-alpha to PA-plasmin system and MMP-9. The extracellular matrix of dental pulp is more strongly damaged through IL-6 and TNF-alpha produced in progress of inflammation.

细胞外基质（ECM）在细胞的生长和变性中起着重要作用。由于感染和宿主免疫反应，ECM被基质金属蛋白酶（MMP）破坏。纤溶酶是由纤溶酶原激活剂 (PA) 有限降解纤溶酶原而产生的，可激活潜在的 MMP 并刺激激肽的合成。炎症细胞因子如IL-1β、IL-6和TNF-α是由细菌或病毒的入侵合成的。检查各种炎症细胞因子对人牙髓细胞中 PA/纤溶酶系统的影响，并与对人牙龈细胞的影响进行比较，以表征牙髓炎中的牙髓细胞。取自同一供体的牙髓和牙龈细胞在含10%胎牛血清的α-MEM中、5%CO_2、37℃空气中培养。将处于汇合阶段的两种细胞在含有 2% PCS 的培养基中孵育 24 小时，并用各种细胞因子（IL-1β、IL-6 和 TNF-α）处理。使用荧光底物测量条件培养基中的PA活性。通过RT-PCR分析tPA mRNA的表达。 IL-6和TNF-α共同孵育时，牙髓细胞的PA活性高于牙龈细胞，且tPA mRNA的表达更强。炎症细胞因子(IL-1β、IL-6和TNF-α)通过增强tPA基因表达和MMP-2活性来刺激PA活性，而这些细胞因子刺激MMP-9激活tPA，因此炎症细胞因子可能通过刺激人牙髓细胞的PA/纤溶酶系统参与细胞外基质降解。牙髓细胞的IL-6和TNF-α对PA-纤溶酶系统和MMP-9的反应与牙龈细胞不同。炎症过程中产生的IL-6和TNF-α更强烈地损害牙髓的细胞外基质。