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Eye tissue damage and clinical application of intra vitreous injection

眼组织损伤及玻璃体内注射的临床应用

基本信息

批准号：
23791995
负责人：
YAMASHITA Takehiro
金额：
$ 2.66万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2011
资助国家：
日本
起止时间：
2011 至 2012
项目状态：
已结题

项目摘要

Retinal pigment epithelial (RPE) cells form a blood-ocular barrier, and their polarized property is crucial for maintaining the barrier functions. Tumor necrosis factor alpha (TNF-α), a major pleotropic inflammatory cytokine that disrupts the barrier function and eventual angiogenesis, is expressed in the choroidal neovascularizations of age-related macular degeneration eyes. Thus, it most likely plays an important role in the progression of the disease. The purpose of this study was to compare the effects of TNF-αon the barrier function of polarized RPE cells. Non-polarized RPE cells were used as negative controls. Isolated porcine RPE cells were seeded on Transwell^ membranes. The polarization of the RPE cells was determined by their high transepithelial electrical resistance and by their differential secretion of vascular endothelial growth factor (lower layer/upper layer >2.5X). Polarized RPE cells were incubated with 10 ng/ml of TNF-αand the TER was measured. TNF-αsignificantly decreased the TER of polarized RPE cells by 17.6 ± 2.7% (P < 0.001) of the control at 24 h and that of non-polarized RPE cells by 5.4 ± 6.5% (P = 0.401). The p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, blocked the effects of TNF-αof decreasing the TER. Cell junction-related molecules, e.g., ZO-1, located between cells in control RPE cells, were disassembled by TNF-α, and this breakdown was suppressed by SB203580 in polarized RPEs. These results indicate that the breakdown of the RPE barrier function was caused exclusively by TNF-αin polarized RPEs, and TNF-αwas acting through the p38 MAPK pathways. Investigations of polarized RPE cells should be more suitable for in vitro studies of the pathophysiology of retinochoroidal diseases.

视网膜色素上皮（RPE）细胞形成血眼屏障，其极化特性对维持屏障功能至关重要。肿瘤坏死因子α（TNF-α）是一种破坏屏障功能和最终血管生成的主要多效性炎性细胞因子，在年龄相关性黄斑变性眼的脉络膜新生血管中表达。因此，它很可能在疾病的进展中发挥重要作用。本研究旨在比较TNF-α对极化RPE细胞屏障功能的影响。非极化RPE细胞用作阴性对照。将分离的猪RPE细胞接种在Transwell ®膜上。RPE细胞的极化由它们的高跨上皮电阻和它们的血管内皮生长因子的差异分泌（下层/上层>2.5X）确定。将极化的RPE细胞与10 ng/ml TNF-α孵育，并测量TER。TNF-α使极化RPE细胞的TER降低17.6 ± 2.7%（P < 0.001），使非极化RPE细胞的TER降低5.4 ± 6.5%（P = 0.401）。p38丝裂原活化蛋白激酶（MAPK）抑制剂SB 203580可阻断TNF-α降低TER的作用。细胞连接相关分子，例如，ZO-1位于对照RPE细胞的细胞之间，被TNF-α分解，并且在极化RPE中这种分解被SB 203580抑制。这些结果表明，在极化RPE中，RPE屏障功能的破坏完全由TNF-α引起，并且TNF-α通过p38 MAPK途径起作用。极化RPE细胞的调查应更适合于视网膜脉络膜疾病的病理生理学的体外研究。