Institute of Human Anatomy and Embryology, University of Regensburg , Regensburg, Germany .
J Ocul Pharmacol Ther. 2014 Mar-Apr;30(2-3):237-45. doi: 10.1089/jop.2013.0205. Epub 2014 Feb 27.
Abstract Purpose: The trabecular meshwork (TM) outflow pathways of the aqueous humor show an increase in extracellular matrix in patients with primary open-angle glaucoma (POAG). The increase in TM extracellular matrix appears to be caused by transforming growth factor-β signaling and its downstream mediator connective-tissue growth factor (CTGF). Here we studied whether treatment with the prostaglandin F2α analog fluprostenol modulates the CTGF-mediated increase of the TM extracellular matrix.
Human TM cells from 3 different donors were treated with CTGF (50 ng/mL) and/or fluprostenol (10(-6) M and 10(-7) M) and were analyzed by real-time reverse transcription polymerase chain reaction and Western blotting. Cell supernatants of the treated cells were analyzed by zymography.
Treatment with CTGF induced the expression and synthesis of CTGF, fibronectin, collagen type IV and VI, while treatment with fluprostenol alone had no effects. The effects of CTGF were blocked by 1-h pretreatment with fluprostenol in a dose-dependent manner. Treatment with fluprostenol or combined fluprostenol/CTGF induced the activity of matrix metalloproteinase 2 (MMP2) in TM cells, whereas treatment with CTGF alone had no effects on MMP2 activity.
Fluprostenol blocks the fibrotic effects of CTGF on human TM cells and increases the activity of MMP2. Both effects have the distinct potential to attenuate a CTGF-mediated increase in TM extracellular matrix in patients with POAG and any effects on TM outflow resistance that may result from that.
摘要目的：原发性开角型青光眼（POAG）患者的小梁网（TM）房水流出途径的细胞外基质增加。TM 细胞外基质的增加似乎是由转化生长因子-β信号及其下游介质结缔组织生长因子（CTGF）引起的。在这里，我们研究了前列腺素 F2α 类似物氟前列醇的治疗是否调节 CTGF 介导的 TM 细胞外基质增加。方法：用 CTGF（50ng/mL）和/或氟前列醇（10（-6）M 和 10（-7）M）处理来自 3 个不同供体的人 TM 细胞，并通过实时逆转录聚合酶链反应和 Western blot 进行分析。用明胶酶谱法分析处理过的细胞的细胞上清液。结果：CTGF 处理诱导 CTGF、纤连蛋白、IV 型和 VI 型胶原的表达和合成，而单独用氟前列醇处理没有影响。CTGF 的作用被氟前列醇以剂量依赖的方式预处理 1 小时所阻断。氟前列醇或联合氟前列醇/CTGF 处理诱导 TM 细胞中基质金属蛋白酶 2（MMP2）的活性，而单独用 CTGF 处理对 MMP2 活性没有影响。结论：氟前列醇阻断 CTGF 对人 TM 细胞的纤维化作用，并增加 MMP2 的活性。这两种作用都有可能减弱 POAG 患者 CTGF 介导的 TM 细胞外基质增加，并可能由此产生对 TM 流出阻力的任何影响。
