Departments of Ophthalmology, Sapporo Medical University School of Medicine, Japan.
Departments of Ophthalmology, Sapporo Medical University School of Medicine, Japan.
Exp Eye Res. 2021 Apr;205:108489. doi: 10.1016/j.exer.2021.108489. Epub 2021 Feb 12.
3D organoid cultures were used to elucidate the periocular effects of several anti-glaucoma drugs including a prostaglandin F2α analogue (bimatoprost acid; BIM-A), EP2 agonist (omidenepag; OMD) or a Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor (ripasudil; Rip) on Grave's orbitopathy (GO) related orbital fatty tissue. 3D organoids were prepared from GO related human orbital fibroblasts (GHOFs) obtained from patients with GO. The effects of either 100 nM BIM-A, 100 nM OMD or 10 μM Rip on the 3D GHOFs organoids were examined with respect to organoid size, physical properties by a micro-squeezer, and the mRNA expression of extracellular matrix (ECM) proteins including collagen (COL) 1, COL 4, COL 6, and fibronectin (FN), ECM regulatory genes including lysyl oxidase (LOX), Connective Tissue Growth Factor (CTGF) and inflammatory cytokines including interleukin-1β (IL1β) and interleukin-6 (IL6). The size of the 3D GHOFs organoids decreased substantially in the presence of BIM-A, but also increased substantially in the presence of the others (OMD or Rip). The physical stiffness of the 3D GHOFs organoids was significantly decreased by Rip. BIM-A caused significantly the down-regulation of three ECM genes, Col 1, Col 6 and Fn, and two ECM regulatory genes and the up-regulation of IL6. In the presence of OMD, two ECM genes, Col 1 and Fn, and LOX were significantly down-regulated but IL1β and IL6 were significantly up-regulated. In the case of Rip, Col 1, FN and CTGF were significant down-regulated. Our present findings indicate that anti-glaucoma drugs modulate the structures and physical properties 3D GHOFs organoids in different manners by modifying the gene expressions of ECM, ECM regulatory factors and inflammatory cytokines. The results indicate that the benefits and demerits of anti-glaucoma medications need to be scrutinized carefully, in cases of patients with GO.
3D 类器官培养物被用于阐明几种抗青光眼药物对眼眶脂肪组织的眼周作用，包括前列腺素 F2α 类似物（比马前列素酸；BIM-A）、EP2 激动剂（omidenepag；OMD）或 Rho 相关卷曲螺旋蛋白激酶（ROCK）抑制剂（ripasudil；Rip）在格雷夫斯眼病（GO）相关眼眶纤维脂肪组织中的作用。3D 类器官由从 GO 患者中获得的 GO 相关人眼眶成纤维细胞（GHOFs）制备。使用微挤压仪检查了 100 nM BIM-A、100 nM OMD 或 10 μM Rip 对 3D GHOFs 类器官的大小、物理特性的影响，以及细胞外基质（ECM）蛋白包括胶原蛋白（COL）1、COL4、COL6 和纤维连接蛋白（FN）、ECM 调节基因包括赖氨酰氧化酶（LOX）、结缔组织生长因子（CTGF）和炎症细胞因子包括白细胞介素-1β（IL1β）和白细胞介素-6（IL6）的 mRNA 表达。在 BIM-A 存在下，3D GHOFs 类器官的体积显著减小，但在其他（OMD 或 Rip）存在下也显著增大。Rip 显著降低了 3D GHOFs 类器官的物理硬度。BIM-A 导致三个 ECM 基因 COL1、COL6 和 Fn 以及两个 ECM 调节基因的下调和 IL6 的上调。在 OMD 存在下，两个 ECM 基因 COL1 和 Fn 以及 LOX 显著下调，但 IL1β 和 IL6 显著上调。在 Rip 的情况下，COL1、FN 和 CTGF 显著下调。我们目前的研究结果表明，抗青光眼药物通过调节细胞外基质、细胞外基质调节因子和炎症细胞因子的基因表达，以不同的方式调节 3D GHOFs 类器官的结构和物理特性。结果表明，在 GO 患者中，需要仔细审查抗青光眼药物的益处和弊端。
