肿瘤坏死因子-α和白细胞介素-1β介导的肾近端小管细胞中基质金属蛋白酶-9和金属蛋白酶组织抑制因子-1的调控

TNF-alpha and IL-1beta-mediated regulation of MMP-9 and TIMP-1 in renal proximal tubular cells.

作者信息

Nee Larine E, McMorrow Tara, Campbell Eric, Slattery Craig, Ryan Michael P

机构信息

Department of Pharmacology, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.

出版信息

Kidney Int. 2004 Oct;66(4):1376-86. doi: 10.1111/j.1523-1755.2004.00900.x.

DOI:10.1111/j.1523-1755.2004.00900.x

PMID:15458430

Abstract

BACKGROUND

Tubulointerstitial fibrosis is a morphologic hallmark of chronic kidney disease and is a key factor in the prediction of progression to end-stage renal failure. Disruption of tubular basement membrane and interstitial extracellular matrix (ECM) via cytokine-induced alterations in matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs) may be an important mechanism in this process. The presence of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) and their effects on proximal tubular cells may be critical in this process.

METHODS

Human proximal tubular cells were cultured in hormonally defined medium. Cells at 80% confluency were exposed to TNF-alpha (0.1 to 100 ng/mL) or IL-1beta (0.1 to 100 ng/mL) or a combination of both for 48 hours. Activity and expression of MMP-9 was examined by gelatin zymography and Western blot analysis. TIMP-1 expression was analyzed by Western blotting. Signaling through cytokine receptors, protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) pathways was investigated.

RESULTS

TNF-alpha but not IL-1beta resulted in a dose-dependent increase in the latent form of MMP-9. TIMP-1 was decreased by treatment with either TNF-alpha or IL-1beta. Cotreatment with IL-1beta abolished the induction of MMP-9 but augmented the inhibition of TIMP-1 in the presence of TNF-alpha. Inhibition of PKC provided evidence of the importance of this pathway in mediating the cytokine-induced suppression of TIMP-1 in human kidney (HK-2) cells. Activation of the extracellular signal-regulated protein kinase (ERK1/2) MAPK mediated the up-regulation of MMP-9 by TNF-alpha whereas p38 was found to be involved in the IL-1beta-mediated inhibition of TNF-alpha-stimulated MMP-9.

CONCLUSION

The differential effects of TNF-alpha and IL-1beta on proximal tubular MMP-9 and TIMP-1 expression are mediated through the TNF-RI, the IL-1-RI and the different signaling pathways of PKC, ERK1/2, and p38 MAPK. These findings may provide new insights into the role of proinflammatory cytokines TNF-alpha and IL-1beta in the development and possible therapeutic intervention in tubulointerstitial fibrosis.

摘要

背景

肾小管间质纤维化是慢性肾脏病的形态学标志，也是预测进展至终末期肾衰竭的关键因素。细胞因子诱导基质金属蛋白酶（MMPs）和金属蛋白酶组织抑制剂（TIMPs）发生改变，进而破坏肾小管基底膜和间质细胞外基质（ECM），这可能是该过程中的一个重要机制。促炎细胞因子肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的存在及其对近端肾小管细胞的作用在这一过程中可能至关重要。

方法

人近端肾小管细胞在激素限定培养基中培养。将80%汇合的细胞暴露于TNF-α（0.1至100 ng/mL）或IL-1β（0.1至100 ng/mL）或两者组合中48小时。通过明胶酶谱法和蛋白质印迹分析检测MMP-9的活性和表达。通过蛋白质印迹分析TIMP-1的表达。研究了通过细胞因子受体、蛋白激酶C（PKC）和丝裂原活化蛋白激酶（MAPK）途径的信号传导。

结果

TNF-α而非IL-1β导致MMP-9潜伏形式呈剂量依赖性增加。用TNF-α或IL-1β处理均可降低TIMP-1。IL-1β与TNF-α联合处理可消除MMP-9的诱导，但在TNF-α存在的情况下增强了对TIMP-1的抑制。抑制PKC证明了该途径在介导细胞因子诱导的人肾（HK-2）细胞中TIMP-1抑制中的重要性。细胞外信号调节蛋白激酶（ERK1/2）MAPK的激活介导了TNF-α对MMP-9的上调，而发现p38参与了IL-1β介导的对TNF-α刺激的MMP-9的抑制。