Axonal Protection by Netarsudil, a ROCK Inhibitor, Is Linked to an AMPK-Autophagy Pathway in TNF-Induced Optic Nerve Degeneration
| Autor: | Yasushi Kitaoka, Kana Sase, Chihiro Tsukahara, Naoki Fujita, Ibuki Arizono, Jiro Kogo, Naoto Tokuda, Hitoshi Takagi |
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| Rok vydání: | 2022 |
| Předmět: |
Male
AMPK autophagy netarsudil Thiophenes AMP-Activated Protein Kinases Benzoates Sequestosome-1 Protein LC3 Animals Enzyme Inhibitors Rats Wistar Protein Kinase Inhibitors rho-Associated Kinases Eye Movements Strabismus Amblyopia and Neuro-Ophthalmology Tumor Necrosis Factor-alpha Biphenyl Compounds p62 Optic Nerve Immunohistochemistry Axons Rats Microscopy Electron Pyrimidines Pyrones Intravitreal Injections Nerve Degeneration beta-Alanine Pyrazoles Microtubule-Associated Proteins |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 |
| Popis: | Purpose Netarsudil, a Rho kinase inhibitor with norepinephrine transport inhibitory effect, lowers intraocular pressure, however, its effect on axon damage remains to be elucidated. The aim of the current study was to investigate the effect of netarsudil on TNF-induced axon loss and to examine whether it affects phosphorylated-AMP-activated kinase (p-AMPK) and autophagy in the optic nerve. Methods Intravitreal administration of TNF or TNF with netarsudil was carried out on rats and quantification of axon number was determined. Electron microscopy determined autophagosome numbers. Localization of p-AMPK expression was examined by immunohistochemistry. The changes in p62, LC3-II, and p-AMPK levels were estimated in the optic nerve by immunoblot analysis. The effect of an AMPK activator A769662 or an AMPK inhibitor dorsomorphin on axon number was evaluated. Results Morphometric analysis revealed apparent protection by netarsudil against TNF-induced axon degeneration. Netarsudil increased autophagosome numbers inside axons. Netarsudil treatment significantly upregulated optic nerve LC3-II levels in both the TNF-treated eyes and the control eyes. Increased p62 protein level induced by TNF was significantly ameliorated by netarsudil. The netarsudil administration alone lessened p62 levels. Netarsudil significantly upregulated the optic nerve p-AMPK levels. A769662 exhibited obvious axonal protection against TNF-induced damage. A769662 treatment upregulated LC3-II levels and the increment of p62 level induced by TNF was significantly ameliorated by A769662. Immunohistochemical analysis revealed that p-AMPK is present in axons. Netarsudil-mediated axonal protection was significantly suppressed by dorsomorphin administration. Conclusions Netarsudil upregulated p-AMPK and autophagy. Netarsudil-mediated axonal protection may be associated with upregulated p-AMPK. |
| Databáze: | OpenAIRE |
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