Abstract
Carbohydrate metabolism changes during cellular senescence. Cytosolic malate dehydrogenase (MDH1) catalyzes the reversible reduction of oxaloacetate to malate at the expense of reduced nicotinamide adenine dinucleotide (NADH). Here, we show that MDH1 plays a critical role in the cellular senescence of human fibroblasts. We observed that the activity of MDH1 was reduced in old human dermal fibroblasts (HDFs) [population doublings (PD) 56], suggesting a link between decreased MDH1 protein levels and aging. Knockdown of MDH1 in young HDFs (PD 20) and the IMR90 human fibroblast cell line resulted in the appearance of significant cellular senescence features, including senescence-associated β-galactosidase staining, flattened and enlarged morphology, increased population doubling time, and elevated p16INK4A and p21CIP1 protein levels. Cytosolic NAD/NADH ratios were decreased in old HDFs to the same extent as in MDH1 knockdown HDFs, suggesting that cytosolic NAD depletion is related to cellular senescence. We found that AMP-activated protein kinase, a sensor of cellular energy, was activated in MDH1 knockdown cells. We also found that sirtuin 1 (SIRT1) deacetylase, a controller of cellular senescence, was decreased in MDH1 knockdown cells. These results indicate that the decrease in MDH1 and subsequent reduction in NAD/NADH ratio, which causes SIRT1 inhibition, is a likely carbohydrate metabolism-controlled cellular senescence mechanism.
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Abbreviations
- MDH1:
-
Cytosolic malate dehydrogenase
- MDH2:
-
Mitochondrial malate dehydrogenase
- NAD:
-
Nicotinamide adenine dinucleotide
- NADH:
-
Reduced nicotinamide adenine dinucleotide
- AMPK:
-
AMP-activated protein kinase
- SIRT1:
-
Sirtuin (silent mating type information regulation 2 homolog) 1
- HIC1:
-
Hypermethylated in cancer 1
- HDFs:
-
Human dermal fibroblasts
- PD:
-
Population doublings
- ROS:
-
Reactive oxygen species
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Acknowledgments
This study was supported by grants from National Research Foundation of Korea (20110030133, 20110018053, and 20110027762, K.-S. K.) and a grant from the National R&D Program for Cancer Control, Ministry of Health & Welfare, Republic of Korea (12201901-14924, K.-S. K.). The authors thank Hong-Duk Youn for reagents and valuable comments.
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Seung-Min Lee and So Hee Dho contributed equally to this study.
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Lee, SM., Dho, S.H., Ju, SK. et al. Cytosolic malate dehydrogenase regulates senescence in human fibroblasts. Biogerontology 13, 525–536 (2012). https://doi.org/10.1007/s10522-012-9397-0
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DOI: https://doi.org/10.1007/s10522-012-9397-0