Metformin Alleviates Inflammation and Induces Mitophagy in Human Retinal Pigment Epithelium Cells Suffering from Mitochondrial Damage
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Cell Treatments
2.3. Cell Viability Assays
2.4. Enzyme-Linked Immunosorbent Assay (ELISA)
2.5. Western Blot
2.6. Real-Time Polymerase Chain Reaction (PCR)
2.7. Immunofluorescence
2.8. MitoTracker Red Staining
2.9. Mitochondrial and Total Cellular ROS Levels
2.10. Statistical Analysis
3. Results
3.1. ARPE-19 Cells Remain Viable after an Exposure up to 40 mM Metformin
3.2. Metformin Reduces the Levels of Proinflammatory Cytokines IL-6 and IL-8 in ARPE-19 Cells with Induced Mitochondrial Damage
3.3. Anti-Inflammatory Effects of Metformin Are Not Mediated by the Regulation of NF-κB or MAP Kinases
3.4. Metformin Pre-Treatment Prevents Antimycin A-Induced Loss of the Mitochondrial Membrane Potential in ARPE-19 Cells
3.5. Metformin Affects the Levels of Autophagy Marker Proteins in ARPE-19 Cells
3.6. Metformin Induced Mitophagy in ARPE-19 Cells
3.7. Metformin Selectively Reduces Antimycin A-Induced Mitochondrial ROS Production in ARPE-19 Cells
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Forward | Reverse |
---|---|---|
SQSTM1 | 5′-GGA GCA GAT GAG GAA GAT CG-3′ | 3′-CTT CGG ATT CTG GCA TCT GT-5′ |
MAP1LC3B | 5′-GCA GCA TCC AAC CAA AAT CC-3′ | 3′-CAT TGA GCT GTA AGC GCC TTC T-5′ |
MTOR | 5′-AGC ATC GGA TGC TTA GGA GTG G-3′ | 3′-CAG CCA GTC ATC TTT GGA GAC C-5′ |
NFE2L2 | 5′-AAA TTG AGA TTG ATG GAA CAG CGA A-3′ | 3′-TAT GGC CTG GCT TAC ACA TTC A-5′ |
GAPDH | 5′-GAT CAT CAG CAA TGC CTC CT-3′ | 3′-GGC CAT CCA CAG TCT TCT G-5′ |
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Toppila, M.; Ranta-aho, S.; Kaarniranta, K.; Hytti, M.; Kauppinen, A. Metformin Alleviates Inflammation and Induces Mitophagy in Human Retinal Pigment Epithelium Cells Suffering from Mitochondrial Damage. Cells 2024, 13, 1433. https://doi.org/10.3390/cells13171433
Toppila M, Ranta-aho S, Kaarniranta K, Hytti M, Kauppinen A. Metformin Alleviates Inflammation and Induces Mitophagy in Human Retinal Pigment Epithelium Cells Suffering from Mitochondrial Damage. Cells. 2024; 13(17):1433. https://doi.org/10.3390/cells13171433
Chicago/Turabian StyleToppila, Maija, Sofia Ranta-aho, Kai Kaarniranta, Maria Hytti, and Anu Kauppinen. 2024. "Metformin Alleviates Inflammation and Induces Mitophagy in Human Retinal Pigment Epithelium Cells Suffering from Mitochondrial Damage" Cells 13, no. 17: 1433. https://doi.org/10.3390/cells13171433