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Published on: December 11, 2020
circ_EPB41 Modulates the miR-15a-5p/Bcl-2 Pathway to Alleviate Oxidative Stress Damage in Lens Epithelial Cells
Jie Fu1, Shiyao Huang1, Shaodan Hu1
1Department of Ophthalmology, Shaoxing TCM Hospital Affiliated to Zhejiang Chinese Medical University, Shaoxing, China.
Purpose:
This study aimed to investigate the role of circ_EPB41 in regulating oxidative stress-induced apoptosis in lens epithelial cells by targeting the miR-15a-5p/Bcl-2 pathway, highlighting its potential significance in age-related cataract treatment. Growing evidence suggests that circular RNAs and microRNAs play critical roles in age-related cataract development, making circ_EPB41 a promising target.
Methods:
SRA01/04 cells were exposed to 200 µM H2O2 for 24 h to induce oxidative damage and model age-related cataract. Cell viability and apoptosis were assessed using MTT assays and flow cytometry, respectively. The interaction between circ_EPB41 and miR-15a-5p was analyzed through bioinformatics tools and dual luciferase reporter assays. Additionally, the expression levels of Bax, Bcl-2, miR-15a-5p, and circ_EPB41 were measured using RT-qPCR and Western blot analysis.
Results:
H2O2-induced SRA01/04 cells showed down-regulation of circ_EPB41 and up-regulation of miR-15a-5p. Circ_EPB41 was found to interact with miR-15a-5p, negatively regulating its expression. Furthermore, miR-15a-5p mimics reversed the effects of circ_EPB41 on oxidative stress in SRA01/04 cells, as evidenced by reduced cell viability, increased apoptosis, elevated Bax levels, and decreased Bcl-2 expression. In contrast, miR-15a-5p inhibitors alleviated H2O2-induced oxidative damage by upregulating Bcl-2 expression.
Conclusion:
This study is the first to demonstrate that up-regulation of circ_EPB41 protects lens epithelial cells from oxidative stress-induced apoptosis through the miR-15a-5p/Bcl-2 pathway. These findings suggest that circ_EPB41 may hold potential as a novel therapeutic target for age-related cataract treatment.
Insights
Circular RNA EPB41 (circ_EPB41) protects against oxidative stress in lens cells by regulating the miR-15a-5p/Bcl-2 pathway. This finding offers a potential new therapeutic strategy for age-related cataract.
Area of Science:
- Molecular Biology
- Cell Biology
- Ophthalmology
Background:
- Age-related cataract is a leading cause of vision impairment.
- Circular RNAs (circRNAs) and microRNAs (miRNAs) are implicated in cataract development.
- circ_EPB41 and miR-15a-5p are potential key players in lens epithelial cell homeostasis.
Purpose of the Study:
- To investigate the role of circ_EPB41 in oxidative stress-induced apoptosis in lens epithelial cells.
- To elucidate the mechanism involving the miR-15a-5p/Bcl-2 pathway.
- To assess the therapeutic potential of circ_EPB41 in age-related cataract.
Main Methods:
- Lens epithelial cells (SRA01/04) were subjected to hydrogen peroxide (H2O2) to induce oxidative stress.
- Cell viability and apoptosis were measured using MTT assays and flow cytometry.
- circ_EPB41 and miR-15a-5p expression, and their interaction, were analyzed using RT-qPCR, Western blot, bioinformatics, and dual luciferase reporter assays.
Main Results:
- H2O2 treatment decreased circ_EPB41 and increased miR-15a-5p expression.
- circ_EPB41 directly targets and inhibits miR-15a-5p.
- Upregulating circ_EPB41 protected cells from oxidative stress by inhibiting apoptosis and modulating Bax/Bcl-2 expression, while inhibiting miR-15a-5p reversed these effects.
Conclusions:
- circ_EPB41 up-regulation confers protection against oxidative stress-induced apoptosis in lens epithelial cells via the miR-15a-5p/Bcl-2 pathway.
- circ_EPB41 represents a promising therapeutic target for age-related cataract.
- This study provides novel insights into the molecular mechanisms underlying cataract formation.
