The effect of TGF-β2 on human trabecular meshwork extracellular proteolytic system
The effect of TGF-β2 on human trabecular meshwork extracellular proteolytic system
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DOI：
10.1016/s0014-4835(03)00220-3
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发表时间：
2003-12-01
影响因子：
3.4
通讯作者：
Lütjen-Drecoll, E
中科院分区：
文献类型：
--
作者：
Fuchshofer, R;Welge-Lussen, U;Lütjen-Drecoll, E
本研究旨在探讨原发性开角型青光眼（POAG）患者房水中的转化生长因子-β 2（TGF-β 2）是否会影响人小梁细胞培养物中基质金属蛋白酶（MMPs）活性的相关因子。用RT-PCR、北方Blot、Western Blot和酶谱法检测TGF-β 2对8例人眼小梁细胞MMP-2、MMP-9、膜型MMP（MTI-MMP）和纤溶酶原激活物抑制剂-1（派-1）表达的影响。还使用免疫组织化学研究了TGF-β 2处理对派-1表达的影响。似乎用TGF-β 2处理显著增加MMP-2的前体形式的表达，而活性形式未检测到。MMP-9和MTI-MMP表达不受TGF-β 2处理的影响。然而，派-1表达显著增加。为了研究MMP-2的前体形式向活性形式的转化是否被派-1抑制，使用酶谱法研究了用TGF-β 2和派-1中和抗体处理对MMP-2的影响。使用该治疗方案，MMP-2的活性形式清晰可见，表明TGF-β 2增强派-1表达降低了NIMP活性。通过升高TGF-β 2水平来抑制MMP活性可能有助于青光眼小梁网中ECM的增加。(C)2003爱思唯尔有限公司。保留所有权利。
Our study aimed to investigate whether transforming growth factor-beta2 (TGF-beta2), increased in the aqueous humor of eyes with primary open angle glaucoma (POAG), can affect factors responsible for the activity of matrix metalloproteinases (MMPs) in human trabecular cell cultures. With this goal in mind cultures of human trabecular meshwork (hTM) cells derived from 8 donors were treated with TGF-beta2 for 24, 36 and 48 hr. Influence of TGF-beta2 on expression of MMP-2, MMP-9, membrane type 1-MMP (MTI-MMP) and plasminogen activator inhibitor-1 (PAI-1) was examined using RT-PCR, Northern Blot, Western Blot and zymography. The influence of TGF-beta2 treatment on PAI-1 expression was also investigated using immunohistochemistry. It appeared that treatment with TGF-beta2 significantly increased expression of the proform of MMP-2, whereas the active form was not detectable. MMP-9 and MTI-MMP expression were not influenced by TGF-beta2 treatment. There was, however, a significant increase in PAI-1 expression. To investigate whether transformation of the proform of MMP-2 to the active form was inhibited by PAI-1, the influence of treatment with TGF-beta2 and a PAI-1 neutralizing antibody on MMP-2 was investigated using zymogram method. With this treatment protocol the active form of MMP-2 was clearly visible, indicating that TGF-beta2 enhancement of the PAI-1-expression decreases NIMP activity. Inhibition of MMP activity through elevated levels of TGF-beta2 might contribute to the increase in ECM in the trabecular meshwork of glaucomatous eyes. (C) 2003 Elsevier Ltd. All rights reserved.