Synthesis pattern of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) in human explant organ cultures after treatment with latanoprost and dexamethasone
Synthesis pattern of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) in human explant organ cultures after treatment with latanoprost and dexamethasone
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DOI：
10.1038/eye.2000.92
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发表时间：
2000-06-01
期刊：
影响因子：
3.9
通讯作者：
Ardjomand, N
中科院分区：
文献类型：
--
作者：
El-Shabrawi, Y;Eckhardt, M;Ardjomand, N
目的观察白藜芦醇和糖皮质激素对睫状体、小梁网和视网膜色素上皮细胞基质金属蛋白酶（MMPs）和基质金属蛋白酶组织抑制剂（TIMPs）表达的影响。视网膜色素上皮生长至融合。用拉坦前列素和/或地塞米松处理器官和细胞培养物72小时。采用酶谱法检测MMP-2、MMP-3和MMP-9的活性。结果用10 μ g/ml拉坦前列素处理外植体器官培养物，通过酶谱分析，MMP-2、MMP-3和MMP-9的表达分别上调了36%、112%和156%。地塞米松500 nm使MMP-2分泌量减少13%，MMP-3减少69%。MMP-9在皮质类固醇处理的外植体器官培养物的培养基中检测不到。在地塞米松处理的培养物中加入10 μ g/ml拉坦前列素使MMP-2增加14%，MMP-3增加43%，MMP-9增加49%。使用免疫组织化学，我们发现睫状体内的MMP-2，-3，-9和TIMP-1和-2抗体染色，小梁网中的程度较低。拉坦前列素治疗导致MMP-2增加29%（p < 0.0001），MMP-9增加98%（p < 0.0001），MMP-9增加108%（p < 0.0001）。地塞米松使MMP-2染色减少32%（p < 0.0001），MMP-3染色减少33%（p < 0.0001），MMP-9染色减少83%（p < 0.0001）。在小梁网中几乎没有检测到MMP染色的变化。拉坦前列素治疗和地塞米松均未诱导TIMP分泌的显著变化（p < 0.93）。在未处理的视网膜色素上皮（RPE）细胞的培养基中，检测到的唯一MMP是MMP-2。培养的RPE细胞没有响应任何治疗的变化，在他们的MMP secretory.Conclusion MMP-3和MMP-9和MMP-2通过拉坦前列素在睫状体，但不是小梁网或RPE细胞的轻度诱导的深刻上调。另一方面，皮质类固醇下调MMP在两种组织中的表达。拉坦前列素可逆转皮质类固醇对MMP产生的抑制作用。
Purpose To determine changes in production of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in the ciliary body, the trabecular meshwork and the retinal pigment epithelium induced by both prostaglandins and corticosteroids.Methods Explant organ cultures were removed by a scleral incision 3 mm posterior to the limbus. Retinal pigment epithelium was grown to confluence. Organ and cell cultures were treated with latanoprost and/or dexamethasone for 72 h. The activity of MMP-2, -3 and -9 was assessed using zymography. The synthesis pattern of MMPs and TIMP-1 and -2 was identified using immunostaining.Results Treatment of explant organ cultures with 10 mu g/ml of latanoprost induced a mean upregulation of MMP-2 by 36%, MMP-3 by 112% and MMP-9 by 156% as seen by zymography. Dexamethasone 500 nm reduced the amounts of secreted MMP-2 by 13%, MMP-3 by 69%. MMP-9 was not detectable in the media of corticosteroid-treated explant organ cultures. The addition of 10 mu g/ml of latanoprost to dexamethasone-treated cultures increased MMP-2 by 14%, MMP-3 by 43% and MMP-9 by 49%. Using immunohistochemistry we found staining with antibodies against MMP-2, -3, -9 and TIMP-1 and -2 within the ciliary body, and only to a lesser degree in the trabecular meshwork. Latanoprost treatment caused an increase of 29% in MMP-2 (p < 0.0001), 98% in MMP-9 (p < 0.0001) and 108% in MMP-9 (p < 0.0001). Dexamethasone reduced the staining for MMP-2 by 32% (p < 0.0001), for MMP-3 by 33% (p < 0.0001) and for MMP-9 by 83% (p < 0.0001). Almost no change in staining for MMPs was detectable in the trabecular meshwork. Neither latanoprost treatment nor dexamethasone induced significant changes (p < 0.93) in the secretion of TIMPs. In the media of non-treated retinal pigment epithelium (RPE) cells the only MMP detected was MMP-2. RPE cells in culture did not respond to either treatment with a change in their MMP secretion.Conclusion We detected a profound upregulation of both MMP-3 and MMP-9 and a mild induction of MMP-2 through latanoprost in the ciliary body, but not the trabecular meshwork or RPE cells. Corticosteroids, on the other hand, downregulated MMP expression in both tissues. This inhibiting effect of corticosteroids on MMP production was reversed by latanoprost.