The mitochondrial-Derived peptide humanin protects RPE cells from oxidative stress, senescence, and mitochondrial dysfunction
Open accessAbstract
PURPOSE: To investigate the expression of humanin (HN) in human retinal pigment epithelial (hRPE) cells and its effect on oxidative stress-induced cell death, mitochondrial bioenergetics, and senescence. METHODS: Humanin localization in RPE cells and polarized RPE monolayers was assessed by confocal microscopy. Human RPE cells were treated with 150 μM tert-Butyl hydroperoxide (tBH) in the absence/presence of HN (0.5-10 μg/mL) for 24 hours. Mitochondrial respiration was measured by XF96 analyzer. Retinal pigment epithelial cell death and caspase-3 activation, mitochondrial biogenesis and senescence were analyzed by TUNEL, immunoblot analysis, mitochondrial DNA copy number, SA-β-Gal staining, and p16INK4a expression and HN levels by ELISA. Oxidative stress-induced changes in transepithelial resistance were studied in RPE monolayers with and without HN cotreatment. RESULTS: A prominent localization of HN was found in the cytoplasmic and mitochondrial compartments of hRPE. Humanin cotreatment inhibited tBH-induced reactive oxygen species formation and significantly restored mitochondrial bioenergetics in hRPE cells. Exogenous HN was taken up by RPE and colocalized with mitochondria. The oxidative stress-induced decrease in mitochondrial bioenergetics was prevented by HN cotreatment. Humanin treatment increased mitochondrial DNA copy number and upregulated mitochondrial transcription factor A, a key biogenesis regulator protein. Humanin protected RPE cells from oxidative stress-induced cell death by STAT3 phosphorylation and inhibiting caspase-3 activation. Humanin treatment inhibited oxidant-induced senescence. Polarized RPE demonstrated elevated cellular HN and increased resistance to cell death. CONCLUSIONS: Humanin protected RPE cells against oxidative stress-induced cell death and restored mitochondrial function. Our data suggest a potential role for HN therapy in the prevention of retinal degeneration, including AMD.
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Humanin is able to restore mitochondrial function after oxidative injury in retinal pigment epithelial cells.
“Humanin has demonstrated protective capacity in age-related diseases such as T2DM and Alzheimer's disease (Mahboobi et al., 2014) and is able to restore mitochondrial function after oxidative injury in retinal pigment epithelial cells (Sreekumar et al., 2016).”
From Mitochondrial Activity and Dynamics Changes Regarding Metabolism in Ageing and Obesity (2017) · cites this paper as Sreekumar et al., 2016
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“Humanin has demonstrated protective capacity in age-related diseases such as T2DM and Alzheimer’s disease (Mahboobi et al., 2014) and is able to restore mitochondrial function after oxidative injury in retinal pigment epithelial cells (Sreekumar et al., 2016).”
BackgroundWoodhead, J. S. T., D’Souza, R. F., Hedges, C. P., Wan, J., Berridge, M. V., Cameron-Smith, D., Cohen, P., Hickey, A. J. R., Mitchell, C. J., & Merry, T. L. (2020) · Journal of Applied Physiology · doi:10.1152/japplphysiol.00032.2020 · Cited by 57 · see citing works
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- Authors
- Sreekumar, P. G., Ishikawa, K., Spee, C., Mehta, H. H., Wan, J., Yen, K., Cohen, P., Kannan, R., & Hinton, D. R.
- Affiliations
- Doheny Eye Institute (US)
- University of Southern California (US)
- Corresponding author
- Hinton, David R.
- ORCID
- Venue
- Investigative Opthalmology & Visual Science
- Published
- 2016-03-18
- Volume
- 57
- Issue
- 3
- Pages
- 1238
- ISSN
- 0146-0404 (ISSN-L)
- 1552-5783
- PMC
- PMC4811181
- OpenAlex
- W2319675468
- Cited by
- 179
- References
- 66
- Citations by year
- 2026: 14 · 2025: 9 · 2024: 10 · 2023: 24 · 2022: 24 · 2021: 16 · 2020: 33 · 2019: 17 · 2018: 13 · 2017: 14 · 2016: 5
- FWCI
- 17.36
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Investigative Ophthalmology & Visual Science (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (7r01ag034430-05)
- National Institute on Aging (1R01AG 034430, P01AG034906, # R01AG034430)
- National Eye Institute (P30EY003040, #EY03040, R01EY001545, EY01545)
- National Institute of General Medical Sciences (R01GM090311)
- National Institute of Environmental Health Sciences (R01ES020812)
- Topics
- Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Keywords
- humanin, retinal pigment epithelium, oxidative stress, cellular senescence, mitochondrial dysfunction, mitochondrial biogenesis, mitochondrial DNA copy number, STAT3 phosphorylation, caspase-3 activation, reactive oxygen species, mitochondrial respiration, transepithelial electrical resistance, age-related macular degeneration, retinal degeneration
- MeSH terms
- Cells, Cultured, Humans, Mitochondria, Phosphorylation, RNA, Cellular Senescence, Apoptosis, Oxidative Stress, Gene Expression Regulation, Developmental, Microscopy, Confocal, In Situ Nick-End Labeling, Microscopy, Electron, Transmission, Intracellular Signaling Peptides and Proteins, Retinal Pigment Epithelium
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