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目的探讨肿瘤坏死因子α(TNF-α)或弱氧化修饰低密度脂蛋白(mmLDL)对人脐静脉内皮细胞(HUVECs)PAI-1活性和mRNA表达的影响及其分子机制。方法用发色底物法测定HUVECs培养液中PAI-1的活性。用Northern印迹分析法检测PAI-1基因mRNA的表达情况,并分别用蛋白激酶C穴PKC雪或促分裂原活化蛋白激酶激酶(MAPKK)抑制剂干预上述诱导实验。结果TNF-α或mmLDL作用HUVEC后,PAI-1活性和mRNA基因表达均明显增加。促分裂原活化蛋白激酶-激酶穴MAPKK雪的抑制剂PD98059(60μmol/L)能明显阻断TNF-α穴100U/ml雪或mmLDL穴50μg/ml雪对PAI-1活性和mRNA的诱导,但PKC的抑制剂Staurosporine(10nmol/L)和H7(15μmol/L)无显著阻断作用。结论(1)TNF-α或mmLDL能增强血管内皮细胞PAI-1活性与mRNA表达;穴2雪PAI-1活性提高与其mRNA表达增加有关;(3)MAPK途径可能在TNF-α或mmLDL诱导血管内皮细胞PAI-1表达中起重要作用。
Objective To investigate the effect of tumor necrosis factor α (TNF-α) or low-density lipoprotein (mmLDL) on PAI-1 activity and mRNA expression in human umbilical vein endothelial cells (HUVECs) and its molecular mechanism. Methods The chromogenic substrate method was used to determine the activity of PAI-1 in HUVECs culture medium. The expression of PAI-1 gene mRNA was detected by Northern blot analysis and the above induction experiments were respectively induced by PKC inhibitor of PKC or mitogen-activated protein kinase kinase (MAPKK). Results TNF-α or mmLDL induced a significant increase in PAI-1 activity and mRNA expression after HUVEC. PD98059 (60μmol / L), a mitogen-activated protein kinase kinase inhibitor MAPKK inhibitor of snow, significantly blocked the induction of PAI-1 activity and mRNA by 100U / ml TNF-α or 50LD / PKC inhibitors Staurosporine (10nmol / L) and H7 (15μmol / L) no significant blocking effect. Conclusions (1) TNF-α or mmLDL can enhance PAI-1 activity and mRNA expression in vascular endothelial cells. The increase of PAI-1 activity in cave 2 is related to the increase of mRNA expression. (3) MAPK pathway may be induced by TNF-α or mmLDL Endothelial PAI-1 expression plays an important role.