Celastrol Protects RPE Cells from Oxidative Stress-Induced Cell Death via Activation of Nrf2 Signaling Pathway
Autor: | Fu Shang, Jingyue Zhang, Kewen Zhou, Yeqi Zhou, Lin-Bin Zhou, Xinyu Zhang |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Programmed cell death Cell Survival NF-E2-Related Factor 2 Apoptosis Retinal Pigment Epithelium medicine.disease_cause Protective Agents Biochemistry Small hairpin RNA 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine medicine Humans Viability assay Molecular Biology Gene knockdown Chemistry GCLM Epithelial Cells General Medicine Hydrogen Peroxide Triterpenes Cell biology Oxidative Stress 030104 developmental biology Celastrol Gene Knockdown Techniques Molecular Medicine Pentacyclic Triterpenes Reactive Oxygen Species Oxidation-Reduction 030217 neurology & neurosurgery Oxidative stress Signal Transduction |
Zdroj: | Current molecular medicine. 19(3) |
ISSN: | 1875-5666 |
Popis: | Purpose: Oxidative stress to retinal pigment epithelial (RPE) cells and inflammation are closely related to the pathogenesis of age-related macular degeneration (AMD). Celastrol is a natural compound isolated from the root of Tripterygium wilfordii. Celastrol has been shown to have potent anti-inflammatory and anti-tumor effects in multiple disease models. The objective of this study was to test the anti-oxidative effects of celastrol in RPE cells and to investigate the underlying mechanisms. Methods: ARPE-19 cells were treated with hydrogen peroxide (H2O2) and menadione alone or in combination with celastrol. Cell viability and apoptosis were examined by CCK-8 and TUNEL assay, respectively. The expression of Nrf2 and its target genes, such as GCLM and HO-1 was determined by Western blotting. The knockdown of Nrf2 was done by transfecting ARPE-19 cells with lentivirus encoding shRNA against Nrf2. The knockdown efficiency was determined by real-time quantitative PCR and Western blotting. Results: Treatment of ARPE-19 cells with celastrol significantly attenuated the toxic effects of both H2O2 and menadione. Treatment with celastrol enhanced the expression of transcription factor Nrf2 and its targets, GCLM and HO-1. Knockdown of Nrf2 expression by shRNA partially abolished the protective effects of celastrol. Chemical inhibition of glutathione synthesis by L-buthionine-S,R-sulfoximine (BSO) completely abolished the protective effects of celastrol against H2O2 and menadione-induced damage. However, chemical inhibition of HO-1 activity by ZnPPIX did not reduce the protective effects of celastrol. Conclusion: This study provides evidence that treatment of RPE cells with celastrol shows potent protective effects against oxidative insults via activation of Nrf2 signaling pathway and upregulation of GCLM expression. This finding suggests that celastrol might be used as a potential therapeutic agent for oxidative stress-related eyes diseases, such as AMD. |
Databáze: | OpenAIRE |
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