前列腺素E_2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 E_2 on matrix metalloproteinase-2 activity in synovial cells and its signaling pathway

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。

Objective：To investigate the effects of prostaglandin Ez（PGE2） on matrix metalloproteinase-2（MMP-2） activity in synovial 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 injmy. 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 PGEz,each of seven signal pathway-specific inhibitors was respective｜y used to treat the synovial cells and MMP-2 activity was detected and semi-quan- tiffed. 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-I pathway with Curcumin, and nuclear factor-κB （NF-κB） pathway with Bayl 1-7082 and-7085 dramatically decreased PGE2-induced MMP-2 activity to 43%,25%, 16% and 1 l%,respec- tively（P〈0.01 ）. Conclusions：PGEz-induced MMP-2 activity in synovial cells may aecount 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.