NF-κB活性对TNF-α调节小梁细胞表达MMP-3和MMP-9的影响  

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

作　　者：吴瑜瑜[1] 濮清岚[1] 李超[1] 郭茂生[1] 

机构地区：[1]福建医科大学第二临床医学院眼科,福建泉州362000

出　　处：《眼视光学杂志》2008年第2期100-105,共6页Chinese Journal of Optometry & Ophthalmology

基　　金：福建省自然科学基金资助项目(C0510014);福建医科大学教授学术发展基金资助项目(2006JS0662)

摘　　要：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(matrix metalloproteinases-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)、1ng/ml、10ng/ml、25ng/mlTNF-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(pyrroli-dine 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。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.

关 键 词：细胞培养 小梁细胞 肿瘤坏死因子-α 基质金属蛋白酶 核因子-ΚB 

分 类 号：R775[医药卫生—眼科]