Objective: To explore the effect of ROCK inhibitor Y-27632 on the matrix metalloproteinase 2 and 9 (MMP2 and MMP9) gene expression and activity in tumor necrosis factor α (TNF-α)-treated human umbilical vein endothelial cell (HUVEC).
Methods: HHUVEC was divided into 3 groups, a control group, a TNF-α group, and a TNF-α plus Y-27632 group. The expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), MMP2 and MMP9 were examined by real-time PCR. The MMP2/9 activity was measured by gelatin zymography.
Results: Compared to the control group, the mRNA expressions of ICAM-1, VCAM-1, MMP2 and MMP9 were increased TNF-α-treated cells, which were suppressed by ROCK inhibitor (P<0.01). The MMP2/9 activity was elevated in TNF-α-treated cells, which was reversed by ROCK inhibitor (P<0.05).
Conclusion: ROCK inhibitor can suppress TNF-α-induced inflammation in endothelial cells through down-regulation of MMP2/9.
目的：探讨ROCK通路抑制剂Y-27632对肿瘤坏死因子α(tumor necrosis factor α，TNF-α)诱导人脐静脉内皮细胞(human umbilical vein endothelial cell，HUVEC)基质金属蛋白酶2和9(matrix metalloproteinase 2 and 9，MMP2，MMP9)的表达与活性的影响。方法：体外培养原代HUVEC，分别予以TNF-α (25 ng/mL)、TNF-α+Y-27632(10 μmol/L)处理24 h。 Real-time PCR检测血管细胞黏附分子-1(vascular cell adhesion molecule-1，VCAM-1)、细胞间黏附分子-1(intercellular adhesion molecule-1，ICAM-1)、MMP2和MMP9 mRNA的表达水平；明胶酶谱检测MMP2和MMP9蛋白活性的改变情况。结果：与对照组相比，TNF-α明显上调ICAM-1，VACAM-1，MMP2，MMP9 mRNA的表达(P<0.01)和MMP2，MMP9的蛋白活性(P<0.05)；与TNF-α处理组相比，Y-27632可显著抑制ICAM-1，VCAM-1，MMP2和MMP9 mRNA的表达(P<0.01)，下调MMP2和MMP9的蛋白活性(P<0.05)。结论：Y-27632可以抑制TNF-α诱导的HUVEC炎症反应和MMP2，MMP9 mRNA和蛋白活性水平。.