Skip to main content Skip to main navigation menu Skip to site footer

Hyperglycemia may induce a decrease in myoblast viability: A Preliminary Study

  • Dicky Moch Rizal ,
  • Jajar Setiawan ,
  • Susi Susanti ,
  • Laksmita Dewi Adzillina ,
  • Nandia Septiyorini ,

Abstract

Link of Video Abstract: https://youtu.be/6j-cheXQzQE

 

Background: Hyperglycemia plays a significant role in developing insulin resistance, leading to a continuous decline in metabolism. Uncontrolled long-term hyperglycemia has deleterious effects on muscles, leading to a decline in muscle mass, impaired capillarization, and muscle atrophy. We identified the effect of hyperglycemia on reducing myoblast cell viability through observation using the MTT assay.

Methods: Primary culture of myoblast cells was performed using rat's muscle tissue which was further induced by hyperglycemia by adding D-glucose with concentrations of 10 mM, 25 mM, and 50 mM in the plating media. Cell viability was observed using the MTT assay by identifying the absorbance using an ELISA reader with λ=550-600 nm (595 nm)

Results: There was a significant decrease in cell viability (p=0.000) due to administering various glucose concentrations in the plating media. There is a negative correlation between glucose concentration and myoblast cell viability (P-value = 0.000; t = -8.284; r = -0.926)

Conclusion: Elevated blood sugar levels result in a significant reduction in the viability of myoblast cells, as indicated by the MTT assay. The MTT assay proved invaluable for precisely evaluating myoblast cell viability in the context of hyperglycemia.

References

  1. Mouri M, Badireddy M. Hyperglicemia. Treasure Island: StatPearls [Internet]; 2023. [Available from: https://www.ncbi.nlm.nih.gov/books/NBK430900/] [Accessed on: 23rd March 2023]
  2. KEMENKES. Diabetes: Sufferers in Indonesia could reach 30 million people in 2030 Jakarta: KEMENKES. 2018. [Available from: https://p2ptm.kemkes.go.id/tag/diabetes-penderita-di-indonesia-bisa-mencapai-30-juta-orang-pada-tahun-2030] [Accessed on: 23rd March 2023].
  3. Giri B, Dey S, Das T, Sarkar M, Banerjee J, Dash SK. Chronic hyperglycemia mediated physiological alteration and metabolic distortion leads to organ dysfunction, infection, cancer progression and other pathophysiological consequences: An update on glucose toxicity. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie. 2018;107(1):306-328.
  4. Shannon CE, Merovci A, Fourcaudot M, Tripathy D, Abdul-Ghani M, Wang H, et al. Effects of Sustained Hyperglycemia on Skeletal Muscle Lipids in Healthy Subjects. The Journal of Clinical Endocrinology & Metabolism. 2022;107(8):e3177-e3185.
  5. Venditti P, Di Meo S. The Role of Reactive Oxygen Species in the Life Cycle of the Mitochondrion. International journal of molecular sciences. 2020;21(6):2173.
  6. Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnology annual review. 2005;11(1):127-152.
  7. Stockert JC, Horobin RW, Colombo LL, Blázquez-Castro A. Tetrazolium salts and formazan products in Cell Biology: Viability assessment, fluorescence imaging, and labeling perspectives. Acta histochemica. 2018;120(3):159-167.
  8. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of immunological methods. 1983;65(1-2):55-63.
  9. Ghasemi M, Turnbull T, Sebastian S, Kempson I. The MTT Assay: Utility, Limitations, Pitfalls, and Interpretation in Bulk and Single-Cell Analysis. International journal of molecular sciences. 2021;22(23):12827.
  10. Buranaamnuay K. The MTT assay application to measure the viability of spermatozoa: A variety of the assay protocols. Open veterinary journal. 2021;11(2):251-269.
  11. Vaughan M, Lamia KA. Isolation and Differentiation of Primary Myoblasts from Mouse Skeletal Muscle Explants. Journal of visualized experiments : JoVE. 2019;152(1):e60310.
  12. Kihara Y, Homma J, Takagi R, Ishigaki K, Nagata S, Yamato M. Laminin-221-derived recombinant fragment facilitates isolation of cultured skeletal myoblasts. Regenerative Therapy. 2022;20(1):147-156.
  13. Au - Vaughan M, Au - Lamia KA. Isolation and Differentiation of Primary Myoblasts from Mouse Skeletal Muscle Explants. JoVE. 2019;152(1):e60310.
  14. Rizwan H, Pal S, Sabnam S, Pal A. High glucose augments ROS generation regulates mitochondrial dysfunction and apoptosis via stress signalling cascades in keratinocytes. Life sciences. 2020;241(1):117148.
  15. Badu-Mensah A, Valinski P, Parsaud H, Hickman JJ, Guo X. Hyperglycemia Negatively Affects IPSC-Derived Myoblast Proliferation and Skeletal Muscle Regeneration and Function. Cells. 2022;11(22):3674
  16. Surinlert P, Thitiphatphuvanon T, Khimmaktong W, Pholpramoo C, Tipbunjong C. Hyperglycemia induced C2C12 myoblast cell cycle arrest and skeletal muscle atrophy by modulating sirtuins gene expression in rats. Polish journal of veterinary sciences. 2021;24(4):563-572.
  17. CCRC. MTT assay procedure: Cancer Chemoprevention Research Center (CCRC), Faculty of Pharmacy, Universitas Gadjah Mada. Yogyakarta: CCRC. 2023.
  18. Riss TL, Moravec RA, Niles AL, al. e. ell Viability Assays. Bethesda (MD): Eli Lilly & Company and the National Center; 2016. [Available from: https://www.ncbi.nlm.nih.gov/books/NBK144065/] [Accessed on: 23rd March 2023].
  19. Kamiloglu S, Sari G, Ozdal T, E C. Guidelines for cell viability assays. Food Frontiers. 2020;1(3):332-349.
  20. Feng CQ, Ma WL, Song YB, Guo QY, Wu QH, Zheng WL. Detection of cell apoptosis by MTT assay. Academic Journal of The First Medical College of PLA. 2002;22(3):262-263.
  21. Binjawhar DN, Alhazmi AT, Bin Jawhar WN, MohammedSaeed W, Safi SZ. Hyperglycemia-induced oxidative stress and epigenetic regulation of ET-1 gene in endothelial cells. Frontiers in genetics. 2023;14(1):1167773.
  22. Shaik NA, Shaik JP, Ali S, Imran A, Rao DK. Increased frequency of micronuclei in diabetes mellitus patients using pioglitazone and glimepiride in combination. Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association. 2010;48(12):3432-3435.
  23. Bima AIH, Elsamanoudy AZ, Albaqami WF, Khan Z, Parambath SV, Al-Rayes N, et al. Integrative system biology and mathematical modeling of genetic networks identifies shared biomarkers for obesity and diabetes. Mathematical biosciences and engineering : MBE. 2022;19(3):2310-2329.
  24. Aziz MS, Aamir AU, Khan A, Khan Z, Shah SQ, Safi SZ, et al. Investigation of Klotho G395A and C1818T Polymorphisms and Their Association with Serum Glucose Level and Risk of Type 2 Diabetes Mellitus. Genes. 2022;13(9):1532.
  25. Bima AI, Elsamanoudy AZ, Alamri AS, Felimban R, Felemban M, Alghamdi KS, et al. Integrative global co-expression analysis identifies key microRNA-target gene networks as key blood biomarkers for obesity. Minerva medica. 2022;113(3):532-541.
  26. Kalyani RR, Metter EJ, Egan J, Golden SH, Ferrucci L. Hyperglycemia predicts persistently lower muscle strength with aging. Diabetes care. 2015;38(1):82-90.
  27. Hirata Y, Nomura K, Senga Y, Okada Y, Kobayashi K, Okamoto S, et al. Hyperglycemia induces skeletal muscle atrophy via a WWP1/KLF15 axis. JCI insight. 2019;4(4): e124952
  28. Monaco CMF, Perry CGR, Hawke TJ. Diabetic Myopathy: current molecular understanding of this novel neuromuscular disorder. Current opinion in neurology. 2017;30(5):545-552.
  29. Kalyani RR, Tra Y, Yeh HC, Egan JM, Ferrucci L, Brancati FL. Quadriceps strength, quadriceps power, and gait speed in older U.S. adults with diabetes mellitus: results from the National Health and Nutrition Examination Survey, 1999-2002. Journal of the American Geriatrics Society. 2013;61(5):769-775.
  30. D'Souza DM, Al-Sajee D, Hawke TJ. Diabetic myopathy: impact of diabetes mellitus on skeletal muscle progenitor cells. Frontiers in physiology. 2013;4(1):379.
  31. Peterson JM, Bryner RW, Alway SE. Satellite cell proliferation is reduced in muscles of obese Zucker rats but restored with loading. American journal of physiology Cell physiology. 2008;295(2):C521-C528.
  32. Yu T, Robotham JL, Yoon Y. Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology. Proceedings of the National Academy of Sciences of the United States of America. 2006;103(8):2653-2658.
  33. Yu T, Jhun BS, Yoon Y. High-glucose stimulation increases reactive oxygen species production through the calcium and mitogen-activated protein kinase-mediated activation of mitochondrial fission. Antioxidants & redox signaling. 2011;14(3):425-437.
  34. Yu T, Sheu SS, Robotham JL, Yoon Y. Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovascular research. 2008;79(2):341-351.
  35. Kaikini AA, Kanchan DM, Nerurkar UN, Sathaye S. Targeting Mitochondrial Dysfunction for the Treatment of Diabetic Complications: Pharmacological Interventions through Natural Products. Pharmacognosy reviews. 2017;11(22):128-135.
  36. Artha IMJR, Bhargah A, Dharmawan NK, Pande UW, Triyana KA, Prabawa IPY, et al. High level of individual lipid profile and lipid ratio as a predictive marker of poor glycemic control in type-2 diabetes mellitus. Vascular Health and Risk Management, 2019;1(1):149-157.
  37. Trisnadewi NW, Adiputra IMS, Oktaviani NPW, Suapriyanti PA, Intan Saraswati NLG. Correlation between stress level and family support towards fasting and postprandial glucose level in type 2 diabetes mellitus. Bali Medical Journal. 2020;9(3):811–815.
  38. Wijaya MC, Sari GM, Tinduh D. Hyperglycemia caused reduction of cortical bone thickness in streptozotocin-induced diabetic rat. Bali Medical Journal. 2017;6(1): 161–163.
  39. Wahjuni S, Gunawan IWG, Malindo IYD. The effect of mustard greens (Brassica rapa l.) ethanol extract on blood glucose and malondialdehyde levels of hyperglycemic Wistar rats. Bali Medical Journal. 2019;8(1):35–40.

How to Cite

Rizal, D. M., Setiawan, J., Susanti, S., Adzillina, L. D., & Septiyorini, N. (2023). Hyperglycemia may induce a decrease in myoblast viability: A Preliminary Study. Bali Medical Journal, 12(3), 2715–2720. https://doi.org/10.15562/bmj.v12i3.4663

HTML
0

Total
0

Share

Search Panel

Dicky Moch Rizal
Google Scholar
Pubmed
BMJ Journal


Jajar Setiawan
Google Scholar
Pubmed
BMJ Journal


Susi Susanti
Google Scholar
Pubmed
BMJ Journal


Laksmita Dewi Adzillina
Google Scholar
Pubmed
BMJ Journal


Nandia Septiyorini
Google Scholar
Pubmed
BMJ Journal