Department of Ophthalmology, Asahikawa Medical University, Asahikawa, Hokkaido, Japan.
Department of Ophthalmology, Asahikawa Medical University, Asahikawa, Hokkaido, Japan.
Exp Eye Res. 2023 Sep;234:109571. doi: 10.1016/j.exer.2023.109571. Epub 2023 Jul 18.
Blinking is regarded as mechanical stimulation of fluid shear stress on the corneal epithelial cells. Therefore, we evaluated whether fluid shear stress affects matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in cultured human corneal epithelial cells (HCECs). No other study has shown the influence of fluid shear stress on HCECs regarding mRNA expression and the protein levels of MMPs. Cultured HCECs were exposed to shear stress (0, 1.2, 12 dyne/cm) for 12 and 24 h with the parallel-plate type of flow chamber. Gene expression of MMPs and TIMPs was measured by real-time polymerase reaction. Concentrations of MMP-1 and MMP-9 in cell lysates were determined using bead-based amplified luminescent proximity homogenous assay-linked immunosorbent assay. The expression of MMP-9 and MMP-1 in HCECs exposed to low and high flow for 12 and 24 h, respectively, increased significantly compared with those under static conditions. The expression of MMP-9 in the cells exposed to high flow for 24 h increased significantly compared with those under static and low flow conditions. Levels of MMP-9 in cell lysates exposed to fluid flow for 24 h were elevated significantly with increasing shear stress. Fluid shear stress exerted on HCECs affected MMPs, which was associated with inflammation and pathogenesis. Mechanical stress induced by blinking might influence expression of MMPs on the ocular surface. Further studies are warranted to establish the molecular mechanism of shear stress-induced alternations of MMPs.
眨眼被认为是对角膜上皮细胞的流体切应力的机械刺激。因此，我们评估了流体切应力是否会影响培养的人角膜上皮细胞（HCEC）中的基质金属蛋白酶（MMPs）和金属蛋白酶组织抑制剂（TIMPs）。没有其他研究表明流体切应力对 HCEC 的 mRNA 表达和 MMPs 的蛋白水平有影响。用平行板流动室将培养的 HCEC 暴露于 0、1.2 和 12 达因/平方厘米的切应力下 12 和 24 小时。通过实时聚合酶反应测量 MMPs 和 TIMPs 的基因表达。使用基于珠的放大发光亲合测定-酶联免疫吸附测定法测定细胞裂解物中 MMP-1 和 MMP-9 的浓度。与静态条件相比，暴露于低和高流量 12 和 24 小时的 HCEC 中 MMP-9 和 MMP-1 的表达分别显著增加。暴露于高流量 24 小时的细胞中 MMP-9 的表达与静态和低流量条件相比显著增加。暴露于流体流动 24 小时的细胞裂解物中的 MMP-9 水平随着切应力的增加而显著升高。施加在 HCEC 上的流体切应力会影响 MMPs，这与炎症和发病机制有关。眨眼引起的机械应力可能会影响眼表面的 MMPs 表达。需要进一步的研究来建立剪切应力诱导的 MMP 变化的分子机制。
