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Updated: Aug 31, 2025
Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells
Published on: December 11, 2020
Downregulation of Inflammatory Response via Nrf2/Trx1/TXNIP Axis in Oxidative Stress-Induced ARPE-19 Cells and Mouse
Qian Yang1, Wenting Cai1, Huizi Jin1
1Department of Ophthalmology, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai 200072, China.
Aim:
Chronic inflammation is crucial for age-related macular degeneration (AMD) pathogenesis. However, the mechanism involved in activating inflammation remains unclear. This study is aimed at investigating whether nuclear factor erythrocyte-associated factor 2 (Nrf2) negatively regulated the Nod-like receptor protein 3 (NLRP3) inflammasomes through the thioredoxin 1 (Trx1)/thioredoxin interaction protein (TXNIP) complex.
Methods:
We determined the optimal hydrogen peroxide (H2O2) concentration, time, and changes in reactive oxygen species (ROS) levels. We also constructed animal models using blue LED irradiation. Then, the expression of Nrf2, TXNIP, Trx1, NLRP3, and inflammation-related factors and proteins, along with the changes in retinal thickness and functional status, was analyzed.
Results:
The oxidative stress model was established after 1 h intervention with 100 μM H2O2. Nrf2 reduced ROS production, protected the ultrastructure of mitochondria, increased the thickness of the ONL layer, and increased the amplitude of a- and b-wave amplitudes in ERG. Trx1 knockdown increased the production of ROS, damaged the ultrastructure of mitochondria, reduced the thickness of the other ONL layer, and reduced the amplitudes of a- and b-waves in the electroretinogram (ERG). Thus, TXNIP in the cytoplasm activated the inflammasomes.
Conclusions:
Nrf2 showed antioxidant and anti-inflammatory activity in the H2O2-induced cell stress model and blue LED-induced retinal light damage model. TXNIP transferred from the nucleus to the cytoplasm, activated NLRP3, and aggravated the retinal injury in both the cell stress model and the animal blue LED model. In contrast, Trx1 knockout promoted this process. This study revealed the possible role of the thioredoxin system in developing AMD while also providing newer insights for the future treatment of AMD.
Insights
Nuclear factor erythrocyte-associated factor 2 (Nrf2) reduces inflammation in age-related macular degeneration (AMD) by inhibiting the Nod-like receptor protein 3 (NLRP3) inflammasome. This pathway involves the thioredoxin 1 (Trx1)/thioredoxin interaction protein (TXNIP) complex, offering new therapeutic targets for AMD.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- Chronic inflammation is a key factor in age-related macular degeneration (AMD) pathogenesis.
- The precise mechanisms activating inflammation in AMD remain incompletely understood.
- Nuclear factor erythrocyte-associated factor 2 (Nrf2) is a transcription factor known for its antioxidant and anti-inflammatory roles.
Purpose of the Study:
- To investigate the role of Nrf2 in regulating Nod-like receptor protein 3 (NLRP3) inflammasomes.
- To elucidate the involvement of the thioredoxin 1 (Trx1)/thioredoxin interaction protein (TXNIP) complex in this regulatory pathway.
- To explore potential therapeutic targets for AMD based on these molecular interactions.
Main Methods:
- Establishment of oxidative stress and blue light-induced retinal damage models.
- Determination of optimal hydrogen peroxide (H2O2) concentrations and reactive oxygen species (ROS) levels.
- Analysis of Nrf2, TXNIP, Trx1, NLRP3 expression, and retinal structural/functional changes.
Main Results:
- Nrf2 activation reduced ROS production and protected mitochondrial ultrastructure in oxidative stress models.
- Trx1 knockdown exacerbated ROS production and mitochondrial damage.
- TXNIP translocation to the cytoplasm activated NLRP3 inflammasomes, leading to increased retinal injury.
Conclusions:
- Nrf2 exhibits significant antioxidant and anti-inflammatory effects in models of AMD.
- TXNIP plays a critical role in activating NLRP3 inflammasomes and exacerbating retinal injury.
- The thioredoxin system is implicated in AMD development, suggesting novel therapeutic strategies.
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