Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8641, Japan.
Cell Tissue Res. 2022 Nov;390(2):173-187. doi: 10.1007/s00441-022-03670-5. Epub 2022 Jul 27.
Matrix metalloproteinases (MMPs) and tumor necrosis factor (TNF)-α contribute to the pathogenesis of several ocular diseases. Previous studies have shown that MMP-9 activation plays an important role in capillary degeneration in injured retinas. In this study, we aimed to determine the roles of TNF-α in capillary degeneration and MMP-9 activation in the injured retina. In rats, retinal injury was induced by intravitreal injection of N-methyl-D-aspartic acid (NMDA, 200 nmol) at postnatal day 7. We examined (1) the effects of blocking MMP-9 and TNF-α signaling pathway on capillary degeneration, (2) changes in protein levels and distribution of MMP-9 and TNF-α, and (3) the interaction between MMP-9 and TNF-α in regulating the expression level of each protein in retinas of NMDA-injected eyes. Intravitreal injection of GM6001, an MMP inhibitor, or TNF-α neutralizing antibody (anti-TNF-α Ab) attenuated capillary degeneration in retinas of NMDA-injected eyes. Protein levels of TNF-α increased 2 h after NMDA injection, whereas those of MMP-9 increased 4 h after the injection. Anti-TNF-α Ab suppressed activation of MMP-9 in retinas of NMDA-injected eyes, whereas GM6001 diminished the TNF-α protein expression. Incubation of recombinant TNF-α with supernatants of homogenized retina increased protein levels and activity of MMP-9. These results suggest that TNF-α and MMP-9 collaboratively increase their expression levels in the retina following neurodegeneration, thus leading to retinal capillary degeneration. The cooperative interaction between MMP-9 and TNF-α could be involved in the exacerbation of retinal neurovascular degeneration.
基质金属蛋白酶 (MMPs) 和肿瘤坏死因子 (TNF)-α 参与多种眼部疾病的发病机制。先前的研究表明，MMP-9 的激活在受损视网膜中的毛细血管退化中起重要作用。在这项研究中，我们旨在确定 TNF-α 在受损视网膜中的毛细血管退化和 MMP-9 激活中的作用。在大鼠中，通过在出生后第 7 天向玻璃体腔注射 N-甲基-D-天冬氨酸 (NMDA，200nmol) 诱导视网膜损伤。我们检查了 (1) 阻断 MMP-9 和 TNF-α 信号通路对毛细血管退化的影响，(2) MMP-9 和 TNF-α 的蛋白水平和分布变化，以及 (3) MMP-9 和 TNF-α 之间的相互作用在调节 NMDA 注射眼视网膜中每种蛋白的表达水平。玻璃体腔内注射 MMP 抑制剂 GM6001 或 TNF-α 中和抗体 (抗 TNF-α Ab) 可减轻 NMDA 注射眼视网膜的毛细血管退化。NMDA 注射后 2 小时 TNF-α 蛋白水平增加，而 MMP-9 蛋白水平增加 4 小时后增加。抗 TNF-α Ab 抑制 NMDA 注射眼视网膜中 MMP-9 的激活，而 GM6001 则降低 TNF-α 蛋白表达。重组 TNF-α 与匀浆视网膜上清液孵育可增加 MMP-9 的蛋白水平和活性。这些结果表明，TNF-α 和 MMP-9 在神经退行性变后协同增加视网膜中的表达水平，从而导致视网膜毛细血管退化。MMP-9 和 TNF-α 之间的协同相互作用可能涉及视网膜神经血管退化的加重。
