A Theacrine-Based Supplement Increases Cellular NAD + Levels and Affects Biomarkers Related to Sirtuin Activity in C2C12 Muscle Cells In Vitro

• Published in: Nutrients 2020-12, Vol.12 (12), p.3727
• Main Authors: Mumford, Petey W, Osburn, Shelby C, Fox, Carlton D, Godwin, Joshua S, Roberts, Michael D
• Format: Article
• Language: English
• Subjects: Adenine; Aging; Animals; Biological activity; Biomarkers; Biomarkers - metabolism; Biopsy; Biosynthesis; Brief Report; Cell culture; Cellulose; Cytokines - metabolism; Cytotoxicity; Dietary supplements; Experimentation; Humans; Inflammation; Laboratories; Lymphocytes T; Mitochondria; Mitochondria - metabolism; mRNA; mRNAs; Muscles; Muscles - metabolism; Musculoskeletal system; Myoblasts; Myoblasts - metabolism; Myotubes; NAD - metabolism; NAD - therapeutic use; Nicotinamide; Nicotinamide phosphoribosyltransferase; Nicotinamide Phosphoribosyltransferase - metabolism; Phosphoribosyltransferase; Proteins; RNA, Messenger; Rodentia; Rodents; SIRT1 protein; Sirtuin 1 - metabolism; sirtuins; Sirtuins - metabolism; theacrine; Uric Acid - administration & dosage; Uric Acid - analogs & derivatives
• ISSN: 2072-6643; 2072-6643
• DOI: 10.3390/nu12123727

There is evidence in rodents to suggest that theacrine-based supplements modulate tissue sirtuin activity as well as other biological processes associated with aging. Herein, we examined if a theacrine-based supplement (termed NAD3) altered sirtuin activity in vitro while also affecting markers of mitochondrial biogenesis. The murine C2C12 myoblast cell line was used for experimentation. Following 7 days of differentiation, myotubes were treated with 0.45 mg/mL of NAD3 (containing ~2 mM theacrine) for 3 and 24 h ( = 6 treatment wells per time point). Relative to control (CTL)-treated cells, NAD3 treatments increased ( < 0.05) Sirt1 mRNA levels at 3 h, as well as global sirtuin activity at 3 and 24 h. Follow-up experiments comparing 24 h NAD3 or CTL treatments indicated that NAD3 increased nicotinamide phosphoribosyltransferase (NAMPT) and SIRT1 protein levels ( < 0.05). Cellular nicotinamide adenine dinucleotide (NAD ) levels were also elevated nearly two-fold after 24 h of NAD3 versus CTL treatments ( < 0.001). Markers of mitochondrial biogenesis were minimally affected. Although these data are limited to select biomarkers in vitro, these preliminary findings suggest that a theacrine-based supplement can modulate select biomarkers related to NAD biogenesis and sirtuin activity. However, these changes did not drive increases in mitochondrial biogenesis. While promising, these data are limited to a rodent cell line and human muscle biopsy studies are needed to validate and elucidate the significance of these findings.