Huang Fu-Mei, Yang Shun-Fa, Hsieh Yih-Shou, Liu Chia-Ming, Yang Li-Chiu, Chang Yu-Chao
School of Dentistry, Oral Medicine Center, Chung Shan Medical University Hospital, Taiwan.
Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2004 Mar;97(3):398-403. doi: 10.1016/j.tripleo.2003.10.007.
Matrix metalloproteinases (MMPs) play an important role in pulp tissue destruction. However, the mechanisms and signal transduction pathways involved in the production of MMPs in human pulp cells are not fully understood. The purpose of this study was to investigate the gelatinolytic activity in human pulp cells stimulated with various pharmacological agents.
Human dental pulp cells were cultured using an explant technique obtained from impacted third molars with informed consent of the patients. The effects of p38 inhibitor SB203580, MEK inhibitor U0126, extracellular signal-regulated kinase (ERK) inhibitor PD098059, phosphatidylinositaol 3-kinase (PI3K) inhibitor LY294002, cyclooxygenase-2 (COX-2) inhibitor NS-398, nuclear factor kappa B (NF-kappaB) inhibitor dexamethasone, and tyrosine kinase inhibitor herbimycin A on the production and secretion of MMPs by human pulp cells were determined by gelatin zymography.
The main gelatinase secreted by human pulp cells migrated at 72 kd and represented MMP-2. Minor gelatinolytic bands were also observed at 92 kd regions that correspond to MMP-9. After a 4-day culture period, NS-398, dexamethasone, and herbimycin A were found to depress MMP-2 production (P<.05). The inhibition decreased in an order of dexamethasone >NS-398>herbimycin A. Human pulp cells, however, treated with various pharmacological agents had no effect on the pattern of MMP-9 produced or secreted in either cell extracts or conditioned medium fractions (P>.05).
These observations suggest that NS-398, dexamethasone, and herbimycin A can regulate MMP-2 produced by human pulp cells. The signal transduction pathways COX-2, NF-kappaB, and tyrosine kinase may be involved in the production of MMPs.
基质金属蛋白酶（MMPs）在牙髓组织破坏中起重要作用。然而，人类牙髓细胞中MMPs产生所涉及的机制和信号转导途径尚未完全明确。本研究的目的是调查用各种药理试剂刺激后的人类牙髓细胞中的明胶酶活性。
在获得患者知情同意后，采用从阻生第三磨牙获取的外植体技术培养人类牙髓细胞。通过明胶酶谱法测定p38抑制剂SB203580、MEK抑制剂U0126、细胞外信号调节激酶（ERK）抑制剂PD098059、磷脂酰肌醇3激酶（PI3K）抑制剂LY294002、环氧合酶-2（COX-2）抑制剂NS-398、核因子κB（NF-κB）抑制剂地塞米松以及酪氨酸激酶抑制剂赫曲霉素A对人类牙髓细胞MMPs产生和分泌的影响。
人类牙髓细胞分泌的主要明胶酶在72 kd处迁移，代表MMP-2。在对应于MMP-9的92 kd区域也观察到较小的明胶酶解条带。培养4天后，发现NS-398、地塞米松和赫曲霉素A可抑制MMP-2的产生（P<0.05）。抑制作用按地塞米松>NS-398>赫曲霉素A的顺序降低。然而，用各种药理试剂处理的人类牙髓细胞对细胞提取物或条件培养基组分中产生或分泌的MMP-9模式没有影响（P>0.）。
这些观察结果表明，NS-398、地塞米松和赫曲霉素A可调节人类牙髓细胞产生的MMP-2。信号转导途径COX-2、NF-κB和酪氨酸激酶可能参与MMPs的产生。
