中波紫外线对角质形成细胞MMP-9及TIMP-1的影响

摘要

目的:研究中波紫外线(UVB)照射对HaCaT细胞中基质金属蛋白酶-9(MMP-9)和基质金属蛋白酶抑制剂-1(TIMP-1)mRNA表达的影响.方法:培养HaCaT细胞,绘制细胞生长曲线,经不同剂量(0、30、60、90 mJ/cm2) UVB照射；用MTT方法测定UYB照射后细胞的增殖活性,反转录-聚合酶链反应(RT-PCR)方法测定UVB照射后HaCaT细胞中MMP-9 mRNA和TIMP-1 mRNA的表达.结果:随着照光剂量的增大,细胞的增殖活性逐渐降低,而细胞的增殖抑制率逐渐增加,不同照射剂量各组间光密度(OD)值差异有统计学意义(P＜0.05)；随着照光剂量的增大,MMP-9mRNA的表达逐渐增加,各组之间比较差异均有统计学意义(P＜0.05)；TIMP- 1mRNA的表达逐渐降低,与0mJ/cm2比较差异有统计学意义(P＜0.05).结论:UVB照射可诱导HaCaT细胞损伤和细胞凋亡,MMP-9和TIMP-1可能与光老化的发生有一定关系.%Objective:To investigate effects of ultraviolet B (UVB) irradiation on matrix metalloproteinase (MMP-9) and tissue inhibitor of matrix metalloproteinase (TIMP-1) mRNA expression in immortalized human keratinocyte (HaCaT) cells. Methods: HaCaT cells were cultured. The cell growth curve was drawn. MTT method was used to determine the cell proliferation, and semiquantitative-PCR (RT-PCR) was used to measure the expression levels of MMP-9 mRNA and TIMP-1 mRNA in HaCaT cells after being treated with different doses of UVB irradiation (0, 30, 60 and 90 mJ/cm2). Results: With the UVB irradiation accumulated, there was a decrease in the proliferation of HaCaT cells and an increase in the inhibition ratio of cell proliferation.There were significant differences in values of OD between different groups (P < 0.05). The expression levels of MMP-9 mRNA increased significantly with the increased irradiation exposure (P < 0.05).The expression level of TIMP-1 decreased gradually, which was significantly different compared with that of 0 mJ/cm2 (P < 0.05). Conclusion : UVB irradiation can induce the injury and apoptosis of HaCaT cells. MMP-9 and TIMP-1 may play a role in the pathogenesis of skin photoag-ing.