NF-κB活性对TNF-α调节小梁细胞表达MMP-3和MMP-9的影响

目的探讨核因子-κB（nuclear factor-κB，NF-κB）是否参与肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）调节人眼小梁细胞（human trabecular cells，HTCs）表达基质金属蛋白酶-3（matrix metalloproteinases-3，MMP-3）和基质 金属蛋白酶-9（matrixmetalloproteinases-9，MMP-9）的过程。方法采用RT-PCR法和酶谱分析法（zymography）检测体外培养的人眼小梁细胞经0ng／ml（对照组）、lng／ml、10ng／ml、25ng/ml TNF-α处理24h后，细胞表达MMP-3、MMP-9的量；运用NF-κB的特异抑制剂二硫氨基甲酸吡啶（pyrrolidine dithocarba-mate，PDTC）抑制NF-κB的活化，观察TNF-α对体外培养的HTCsMMP-3，MMP-9表达影响的改变。结果运用RT-PCR法和酶谱分析法研究均发现经浓度为1ng／ml、10ng／ml、25ng／ml的TNF-α处理的细胞在mRNA和上清液中活性蛋白水平均能增加MMP-3、MMP-9的表达，各组mRNA／β-actin吸光度比值和条带酶解量与对照组相比差异均有显著性（P〈0．05）。提前30min加入PDTC，MMP-3、MMP-9的表达量分别较未加入PDTC组表达量明显减少．差异有显著性（P〈0．05）。结论TNF-α可上调MMP-3及MMP-9的表达。PDTC能够下调TNF-α对小梁细胞MMP-3及MMP-9的表达，因此推测NF-κB可能参与MMP-3和MMP-9转录的启动．

The contribution of nuclear factor kappa-B in the modulation of TNF-α on the expression of MMP-3 and MMP-9 in cultured human trabecular cells

Objective To study the effect of NF-κB in the tumor necrosis factor-α （TNF-α） on the expression of matrix metallopreteinases （MMPs） in euhured human trabecular cells （HTCs）. Methods MMP-3 and MMP-9 expressed in cultured HTCs were measured by semi-quantitative RT-PCR and casein/gelatin zymography after treatment with 0 ng/ml （control）, 1 ng/ml, 10 ng/ml, or 25 ng/ml TNF-α for 24 h. PDTC, an inhibitor of nuclear factorkappa B （NF-κB）, was used before HTCs were treated with 10 ng/ ml or 25 ng/ml TNF-α for 24 h. Results The difference in the mean gray scale and MMP-3, MMP-9 activity between the 1 ng/ ml, 10 ng/ml, and 25 ng/ml TNF-α treatment groups and that of the control group was statistically significant. The expression of MMP-3 and MMP-9 decreased when cells were pretreated for 30 minutes with NF-κB inhibitor PDTC （100μmol/L）. Conclusion TNF-α can increase MMP-3 and MMP-9 expression in cultured HTCs. The fact that TNF-α leads to the activation of NF-κB is critical to the TNF-α-stimulated upregnlation of MMP-3 and MMP-9. Therefore, it is likely that compounds that activate the NF-κB pathway would upregnlate the production of MMP-3 and MMP-9 and improve aqueous outflow.