Cartap-induced cytotoxicity in mouse C2C12 myoblast cell line and the roles of calcium ion and oxidative stress on the toxic effects

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 (n = 6 treatment wells per time point). Relative to control (CTL)-treated cells, NAD3 treatments increased (p < 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 (p < 0.05). Cellular nicotinamide adenine dinucleotide (NAD+) levels were also elevated nearly two-fold after 24 h of NAD3 versus CTL treatments (p < 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.

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Corrigendum to “(CTG)n repeats markedly inhibit differentiation of the C2C12 myoblast cell line: Implications for congenital myotonic dystrophy” [Biochim. Biophys. Acta 1453 (1999) 221–229]

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease ◽

10.1016/j.bbadis.2008.03.005 ◽

2008 ◽

Vol 1782 (5) ◽

pp. 362

Author(s):

Satyakam Bhagavati ◽

S.A. Shafiq ◽

Weimin Xu

Keyword(s):

Cell Line ◽

Myotonic Dystrophy ◽

C2c12 Myoblast ◽

Congenital Myotonic Dystrophy ◽

Myoblast Cell Line ◽

Myoblast Cell

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The growth-promoting activity of egg white proteins in the C2C12 myoblast cell line

Animal Science Journal ◽

10.1111/asj.12257 ◽

2014 ◽

Vol 86 (2) ◽

pp. 194-199 ◽

Cited By ~ 11

Author(s):

Wataru Mizunoya ◽

Ayumi Tashima ◽

Yusuke Sato ◽

Ryuichi Tatsumi ◽

Yoshihide Ikeuchi

Keyword(s):

Cell Line ◽

Egg White ◽

C2c12 Myoblast ◽

Egg White Proteins ◽

Myoblast Cell Line ◽

Growth Promoting ◽

Myoblast Cell

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Novel method for fabrication of skeletal muscle construct from the C2C12 myoblast cell line using serum-free medium AIM-V

Biotechnology and Bioengineering ◽

10.1002/bit.22318 ◽

2009 ◽

Vol 103 (5) ◽

pp. 1034-1041 ◽

Cited By ~ 27

Author(s):

Hideaki Fujita ◽

Kazunori Shimizu ◽

Eiji Nagamori

Keyword(s):

Skeletal Muscle ◽

Cell Line ◽

C2c12 Myoblast ◽

Serum Free Medium ◽

Free Medium ◽

Serum Free ◽

Myoblast Cell Line ◽

Novel Method ◽

Myoblast Cell

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A Comparative Study of the Cytotoxic Effects and Oxidative Stress of Gossypol on Bovine Kidney and HeLa Cell Lines

Iranian Jornal of Toxicology ◽

10.32598/ijt.15.3.795.1 ◽

2021 ◽

Vol 15 (3) ◽

pp. 157-164

Author(s):

Mahsa Daneshmand ◽

◽

Jamileh Salar Amoli ◽

Tahereh Ali Esfahani ◽

◽

...

Keyword(s):

Oxidative Stress ◽

Hela Cell ◽

Cell Line ◽

Cell Lines ◽

Toxic Effects ◽

Cytotoxic Effects ◽

Bovine Kidney ◽

Hela Cell Lines ◽

Significant Difference ◽

And Oxidative Stress

Background: Cotton seed is one of the main sources of protein in animal feeds, containing gossypol, which has been shown to have toxic effects. Results reported by various studies also indicate the anti-cancer effects of gossypol on various cell types. However, its toxic effects on human and animal cells have not been fully established. This study was planned to investigate, for the first time, the cytotoxic effects and oxidative stress induced by gossypol on normal Bovine Kidney (BK) and HeLa cell lines, representing typical healthy and cancer cells, respectively. Methods: The BK and HeLa cell lines were treated for 24, 48 or 72 hours with 5, 10 or 20 ppm of gossypol (+/-). The cellular bio-availability and cytotoxicity were measured by MTT assay. The catalase and Malondialdehyde (MDA) levels were also measured to represent the oxidative stress parameters. Results: The percentages of cytotoxicity in BK and HeLa cell lines were calculated at a gossypol concentration of 5, 10 and 20 ppm over 24, 48 or 72 hours of incubation, respectively. The Lethal Concentration 50 (lC50) values were also determined for the two cell lines. No changes in the catalase and lipid peroxidase activities were observed in either cell line. Conclusion: The percentage of the gossypol cytotoxicity was concentration-dependent. By comparing the IC50 in both cell lines using one-way Analysis of Variance (ANOVA) analysis, a significant difference was observed, suggesting that Hela cells were less sensitive to gossypol than the BK cells. Lack of changes in the oxidative stress, as tested by catalase and MDA assays, demonstrated that gossypol did not induce oxidative stress in either cell line.

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Interleukin-6 Induces Myogenic Differentiation via JAK2-STAT3 Signaling in Mouse C2C12 Myoblast Cell Line and Primary Human Myoblasts

International Journal of Molecular Sciences ◽

10.3390/ijms20215273 ◽

2019 ◽

Vol 20 (21) ◽

pp. 5273 ◽

Cited By ~ 8

Author(s):

Paul J. Steyn ◽

Kevin Dzobo ◽

Robert I. Smith ◽

Kathryn H. Myburgh

Keyword(s):

Cell Cycle ◽

Protein Expression ◽

Cell Line ◽

Interleukin 6 ◽

Janus Kinase ◽

Cytokine Signaling ◽

C2c12 Myoblast ◽

Myoblast Cell Line ◽

Myoblast Cell ◽

Stat Pathway

Postnatal muscle growth and exercise- or injury-induced regeneration are facilitated by myoblasts. Myoblasts respond to a variety of proteins such as cytokines that activate various signaling cascades. Cytokines belonging to the interleukin 6 superfamily (IL-6) influence myoblasts’ proliferation but their effect on differentiation is still being researched. The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway is one of the key signaling pathways identified to be activated by IL-6. The aim of this study was to investigate myoblast fate as well as activation of JAK-STAT pathway at different physiologically relevant IL-6 concentrations (10 pg/mL; 100 pg/mL; 10 ng/mL) in the C2C12 mouse myoblast cell line and primary human myoblasts, isolated from eight young healthy male volunteers. Myoblasts’ cell cycle progression, proliferation and differentiation in vitro were assessed. Low IL-6 concentrations facilitated cell cycle transition from the quiescence/Gap1 (G0/G1) to the synthesis (S-) phases. Low and medium IL-6 concentrations decreased the expression of myoblast determination protein 1 (MyoD) and myogenin and increased proliferating cell nuclear antigen (PCNA) expression. In contrast, high IL-6 concentration shifted a larger proportion of cells to the pro-differentiation G0/G1 phase of the cell cycle, substantiated by significant increases of both MyoD and myogenin expression and decreased PCNA expression. Low IL-6 concentration was responsible for prolonged JAK1 activation and increased suppressor of cytokine signaling 1 (SOCS1) protein expression. JAK-STAT inhibition abrogated IL-6-mediated C2C12 cell proliferation. In contrast, high IL-6 initially increased JAK1 activation but resulted in prolonged JAK2 activation and elevated SOCS3 protein expression. High IL-6 concentration decreased interleukin-6 receptor (IL-6R) expression 24 h after treatment whilst low IL-6 concentration increased IL-6 receptor (IL-6R) expression at the same time point. In conclusion, this study demonstrated that IL-6 has concentration- and time-dependent effects on both C2C12 mouse myoblasts and primary human myoblasts. Low IL-6 concentration induces proliferation whilst high IL-6 concentration induces differentiation. These effects are mediated by specific components of the JAK/STAT/SOCS pathway.

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