Research for associations of tension-dependent aqueous outflow regulation by trabecular meshwork with glaucoma pathology.

为了确定与张力依赖相关的因素，我们利用小梁细胞的三维培养建立了压力模型。在该模型中，将胶原蛋白、培养液和细胞混合，然后通过改变pH来稳定。在模型中让细胞迁移和增殖后，称重模型，并对模型中的细胞进行免疫染色观察。共聚焦显微镜观察发现F-肌动蛋白表达减少。考虑到降低F-肌动蛋白的ROCK抑制剂可以降低人的眼压，我们假设这种反应通过与ROCK抑制剂相同的机制来促进眼压的动态平衡。然而，在三维培养中很难接近细胞中的蛋白质。因此，我们评估了小梁细胞对氧化应激的反应，因为我们推测，细胞内对压力应激的反应与对氧化应激的反应具有相似的方面。氧化应激后，小梁细胞Akt-PI3K信号和p38信号被激活。此外，通过抑制这些信号，氧化应激诱导的细胞死亡得到加强。这些新颖的发现为我们理解房水流出的调节机制提供了有价值的线索。

To identify factors associated with the tension-dependency, we developed a pressure model by using three-dimension culture of trabecular meshwork cells. In this model, collagen, medium, and cells were mixed and then were stabilized by changing the pH. After letting cells migrate and proliferate in the model, the model was weighed, and cells in the model were observed by immunostaining. F-actin was found to be decreased by observation with the confocal microscopy. Considering ROCK inhibitors which decrease F-actin could lower intraocular pressure in human, we assumed that this response contributed to the homeostasis of intraocular pressure by using the same mechanism with ROCK inhibitors. It was difficult, however, to approach proteins in cells in the three-dimension culture. We therefore assessed the reaction of trabecular cells against oxidative stress, because we speculated that the intracellular reaction against pressure stress had similar aspects with that against oxidative stress. After exposure to oxidative stress, the Akt-PI3K signaling and the p38 signaling in the trabecular meshwork cells were revealed to be activated. Moreover, oxidative stress-induced cell death was enhanced by inhibiting these signalings. These novel findings provided us a valuable clue to understanding the mechanisms that regulate aqueous humor outflow.