诱导型环氧合酶-2在CD40配体诱导基质金属蛋白酶表达中的作用
rhCD40L induces expression of matrix metalloproteinases in cultured U937 cells by cyclooxygenase-2 pathway
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摘要: 目的: 探讨重组人CD40配体（rhCD40L）诱导U937细胞中基质金属蛋白酶（MMPs）表达的作用及其与诱导型环氧合酶-2(COX-2)途径的关系。方法： 体外培养U937细胞，以不同浓度（0.1、0.2、0.4 mg/L）的rhCD40L和不同浓度（10-5 mol/L、10-4 mol/L、10-3 mol/L）的COX-2特异性抑制剂(NS-398)分别刺激24 h。在加入终浓度为0.4 mg/L rhCD40L的基础上，加入终浓度为10-4 mol/L的NS-398，共同刺激U937细胞24 h后，收集培养上清液，用酶谱法测定MMPs的活性。结果： rhCD40L可使MMP-2和MMP-9的活性明显增加（P<0.01），且呈剂量依赖性；NS-398可抑制MMP-2和MMP-9的活性，且抑制作用随剂量的增加而增强（P<0.05）。结论： rhCD40L以浓度依赖的方式诱导MMPs表达，rhCD40L对MMPs的诱导作用可能与COX-2途径有关。Abstract: AIM: To evaluate matrix metalloproteinase (MMP) expression induced by rhCD40L in cultured U937 cell and possible effects of cyclooxygenase-2 (COX-2). METHODS: U937 cells were treated with rhCD40L and NS-398 in different concentrations, respectively, and on the basis of the treatment with rhCD40L, U937 cells were cocultured with NS-398 and aspirin. The supernatant of the U937 cells cultures was collected. Zymography was performed by SDS-PAGE in 10% gels containing 0.1% (w/v) gelatin (denatured collagen) at 4℃. The activities of MMPs were detected by the grey level of the band. RESULTS: rhCD40L increased the activity of MMPs in a dose-dependent manner, whereas NS-398 significantly inhibited the activity of MMPs. NS-398 significantly inhibited the activity of MMPs induced by rhCD40L. CONCLUSION: rhCD40L can induce U937 cells to secrete MMPs in a dose-dependent manner, which may be related to COX-2 pathway.
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