Axe Médecine Régénératrice, Centre de Recherche du CHU de Québec-Université Laval, Québec City, QC G1S 4L8, Canada.
Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Québec City, QC G1J 1Z4, Canada.
Int J Mol Sci. 2024 Sep 23;25(18):10227. doi: 10.3390/ijms251810227.
The corneal endothelium is responsible for pumping fluid out of the stroma in order to maintain corneal transparency, which depends in part on the expression and activity of sodium-potassium pumps. In this study, we evaluated how physiologic pressure and flow influence transcription, protein expression, and activity of Na+/K+-ATPase. Native and engineered corneal endothelia were cultured in a bioreactor in the presence of pressure and flow (hydrodynamic culture condition) or in a Petri dish (static culture condition). Transcription of was assessed using qPCR, the expression of the α1 subunit of Na+/K+-ATPase was measured using Western blots and ELISA assays, and Na+/K+-ATPase activity was evaluated using an ATPase assay in the presence of ouabain. Results show that physiologic pressure and flow increase the transcription and the protein expression of Na+/K+-ATPase α1 in engineered corneal endothelia, while they remain stable in native corneal endothelia. Interestingly, the activity of Na+/K+-ATPase was increased in the presence of physiologic pressure and flow in both native and engineered corneal endothelia. These findings highlight the role of the in vivo environment on the functionality of the corneal endothelium.
角膜内皮细胞负责将液体从基质中泵出，以维持角膜的透明度，这部分依赖于钠钾泵的表达和活性。在这项研究中，我们评估了生理压力和流量如何影响 Na+/K+-ATPase 的转录、蛋白表达和活性。天然和工程化的角膜内皮细胞在存在压力和流量（流体动力学培养条件）的生物反应器中或在培养皿中（静态培养条件）进行培养。使用 qPCR 评估 的转录，使用 Western blot 和 ELISA 测定 Na+/K+-ATPase 的α1 亚基的表达，并且在存在哇巴因的情况下使用 ATPase 测定法评估 Na+/K+-ATPase 活性。结果表明，生理压力和流量增加了工程化角膜内皮细胞中 Na+/K+-ATPaseα1 的转录和蛋白表达，而在天然角膜内皮细胞中则保持稳定。有趣的是，在生理压力和流量存在的情况下，天然和工程化角膜内皮细胞中 Na+/K+-ATPase 的活性均增加。这些发现强调了体内环境对角膜内皮细胞功能的作用。
