前列腺素E2对滑膜细胞基质金属蛋白酶-2活性的影响及其信号通路研究

Effects of prostaglandin E2 on matrix metalloproteinase-2 activity in synovial cells and its signaling pathway

摘要:

目的：观察前列腺素E2（Prostaglandin E2，PGE2）对滑膜细胞基质金属蛋白酶-2（Matrix metalloproteinase-2，MMP-2）活性的影响及其信号通路的研究，从而探讨PGE2对前交叉韧带（Anterior cruciate ligament，ACL）的损伤或修复可能的影响。方法：用不同浓度的PGE2处理体外培养的滑膜细胞，处理后12、24、48 h和72 h明胶酶谱法检测培养液中MMP-2的活性；再用合适浓度的PGE2单独或分别与7种信号通路特异性抑制剂同时处理滑膜细胞，明胶酶谱法检测MMP-2的活性，并对其进行相对定量。结果：PGE2呈浓度和时间依赖性地增高滑膜细胞MMP-2的活性。p38、ERK、PKA通路抑制剂SB203850、PD98059、KT5720对PGE2诱导滑膜细胞MMP-2活性的增高无明显抑制作用；而JNK、AP-1和核因子-κB（Nuclear factor-κB，NF-κB）通路抑制剂SP600125、姜黄素（Curcumin）和Bay11-7082/7085显著抑制了PGE2诱导MMP-2活性的增高，分别使72 h时 MMP-2的活性降至43%、25%和16%、11%（P<0.01）。结论：PGE2诱导滑膜细胞MMP-2活性的增加可能是ACL损伤后难以自行修复的原因之一。JNK、AP-1和NF-κB通路可能参与调控PGE2对MMP-2活性的诱导；其中以NF-κB作用最显著，可能为ACL损伤的治疗提供一种新思路。

Abstract:

Objective：To investigate the effects of prostaglandin E2（PGE2） on matrix metalloproteinase-2（MMP-2） activity in syn－ovial cells and its signaling pathway so as to help discuss the possible effect of PGE2 on the healing ability of anterior cruciate liga－ment（ACL） after injury. Methods：Cultured synovial cells were treated with different concentrations of PGE2，and gelatin zymography assay was performed to detect MMP-2 activity at 12，24，48 h and 72 h after treatment.In presence of 10 ng/ml PGE2，each of seven signal pathway-specific inhibitors was respectively used to treat the synovial cells and MMP-2 activity was detected and semi-quan－tified. Results：PGE2 significantly elevated MMP-2 activity in synovial cells in a dose and time dependent manner. Inhibition of the p38 pathway with SB203850，ERK pathway with PD98059，and PKA pathway with KT5720 had no specific effect on PGE2-induced MMP-2 activity. However，inhibition of the JNK pathway with SP600125，AP-1 pathway with Curcumin，and nuclear factor-κB（NF-κB） pathway with Bay11-7082 and-7085 dramatically decreased PGE2-induced MMP-2 activity to 43%，25%，16% and 11%，respec－tively（P<0.01）. Conclusions：PGE2-induced MMP-2 activity in synovial cells may account for ACL’s poor healing ability after in－jury and this process might be regulated via JNK，AP-1 and NF-κB pathway. Inhibition of NF-κB pathway has the strongest decreas－ing effect on PGE2-induced MMP-2 activity，which provides a novel strategy for ACL repair.