International Journal of Stem Cells

  • 제16권1호
  • /
  • Pages.78-92
  • /
  • 2023
  • /
  • 2005-3606(pISSN)
  • /
  • 2005-5447(eISSN)
한국줄기세포학회 (Korean Society for Stem Cell Research)
권호 표지
DOI QR코드

DOI QR Code

Synergistic Effect of Hydrogen and 5-Aza on Myogenic Differentiation through the p38 MAPK Signaling Pathway in Adipose-Derived Mesenchymal Stem Cells

  • Wenyong Fei (Department of Sports Medicine, Northern Jiangsu People's Hospital, Clinical Medical College, Yangzhou University) ;
  • Erkai Pang (Department of Sports Medicine, Northern Jiangsu People's Hospital, Dalian Medical University) ;
  • Lei Hou (Department of Sports Medicine, Northern Jiangsu People's Hospital, Dalian Medical University) ;
  • Jihang Dai (Department of Sports Medicine, Northern Jiangsu People's Hospital, Clinical Medical College, Yangzhou University) ;
  • Mingsheng Liu (Department of Sports Medicine, Northern Jiangsu People's Hospital, Dalian Medical University) ;
  • Xuanqi Wang (Department of Sports Medicine, Northern Jiangsu People's Hospital, Dalian Medical University) ;
  • Bin Xie (Department of Sports Medicine, Northern Jiangsu People's Hospital, Dalian Medical University) ;
  • Jingcheng Wang (Department of Sports Medicine, Northern Jiangsu People's Hospital, Clinical Medical College, Yangzhou University)
  • 투고 : 2021.12.10
  • 심사 : 2022.06.13
  • 발행 : 2023.02.28
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Background and Objectives: This study aims to clarify the systems underlying regulation and regulatory roles of hydrogen combined with 5-Aza in the myogenic differentiation of adipose mesenchymal stem cells (ADSCs). Methods and Results: In this study, ADSCs acted as an in vitro myogenic differentiating mode. First, the Alamar blue Staining and mitochondrial tracer technique were used to verify whether hydrogen combined with 5-Aza could promote cell proliferation. In addition, this study assessed myogenic differentiating markers (e.g., Myogenin, Mhc and Myod protein expressions) based on the Western blotting assay, analysis on cellular morphological characteristics (e.g., Myotube number, length, diameter and maturation index), RT-PCR (Myod, Myogenin and Mhc mRNA expression) and Immunofluorescence analysis (Desmin, Myosin and 𝛽-actin protein expression). Finally, to verify the mechanism of myogenic differentiation of hydrogen-bound 5-Aza, we performed bioinformatics analysis and Western blot to detect the expression of p-P38 protein. Hydrogen combined with 5-Aza significantly enhanced the proliferation and myogenic differentiation of ADSCs in vitro by increasing the number of single-cell mitochondria and upregulating the expression of myogenic biomarkers such as Myod, Mhc and myotube formation. The expressions of p-P38 was up-regulated by hydrogen combined with 5-Aza. The differentiating ability was suppressed when the cells were cultivated in combination with SB203580 (p38 MAPK signal pathway inhibitor). Conclusions: Hydrogen alleviates the cytotoxicity of 5-Aza and synergistically promotes the myogenic differentiation capacity of adipose stem cells via the p38 MAPK pathway. Thus, the mentioned results present insights into myogenic differentiation and are likely to generate one potential alternative strategy for skeletal muscle related diseases.

키워드

과제정보

This work was supported by the funds from the young medical key talent project of Jiangsu province (contract number QNRC2016458). Jiangsu Provincial Medical Innovation Team (Grant#CXTDB2017004).

참고문헌

  1. Abmayr SM, Pavlath GK. Myoblast fusion: lessons from flies and mice. Development 2012;139:641-656
  2. Greising SM, Warren GL, Southern WM, Nichenko AS, Qualls AE, Corona BT, Call JA. Early rehabilitation for volumetric muscle loss injury augments endogenous regenerative aspects of muscle strength and oxidative capacity. BMC Musculoskelet Disord 2018;19:173
  3. Aguilar CA, Greising SM, Watts A, Goldman SM, Peragallo C, Zook C, Larouche J, Corona BT. Multiscale analysis of a regenerative therapy for treatment of volumetric muscle loss injury. Cell Death Discov 2018;4:33 Erratum in: Cell Death Discov 2018;5:16
  4. Valencia Mora M, Ruiz Iban MA, Diaz Heredia J, Barco Laakso R, Cuellar R, Garcia Arranz M. Stem cell therapy in the management of shoulder rotator cuff disorders. World J Stem Cells 2015;7:691-699
  5. Hernigou P, Flouzat Lachaniette CH, Delambre J, Zilber S, Duffiet P, Chevallier N, Rouard H. Biologic augmentation of rotator cuff repair with mesenchymal stem cells during arthroscopy improves healing and prevents further tears: a case-controlled study. Int Orthop 2014;38:1811-1818
  6. Gimble JM, Katz AJ, Bunnell BA. Adipose-derived stem cells for regenerative medicine. Circ Res 2007;100:1249-1260
  7. Grottkau BE, Lin Y. Osteogenesis of adipose-derived stem cells. Bone Res 2013;1:133-145
  8. Dai R, Wang Z, Samanipour R, Koo KI, Kim K. Adiposederived stem cells for tissue engineering and regenerative medicine applications. Stem Cells Int 2016;2016:6737345
  9. Xie X, Wang Y, Zhao C, Guo S, Liu S, Jia W, Tuan RS, Zhang C. Comparative evaluation of MSCs from bone marrow and adipose tissue seeded in PRP-derived scaffold for cartilage regeneration. Biomaterials 2012;33:7008-7018
  10. Liu Y, Yan X, Sun Z, Chen B, Han Q, Li J, Zhao RC. Flk-1+ adipose-derived mesenchymal stem cells differentiate into skeletal muscle satellite cells and ameliorate muscular dystrophy in mdx mice. Stem Cells Dev 2007;16:695-706
  11. Abdanipour A, Tiraihi T, Delshad A. Trans-differentiation of the adipose tissue-derived stem cells into neuron-like cells expressing neurotrophins by selegiline. Iran Biomed J 2011;15:113-121
  12. Mizuno H, Zuk PA, Zhu M, Lorenz HP, Benhaim P, Hedrick MH. Myogenic differentiation by human processed lipoaspirate cells. Plast Reconstr Surg 2002;109:199-209; discussion 210-211
  13. McClure MJ, Garg K, Simpson DG, Ryan JJ, Sell SA, Bowlin GL, Ericksen JJ. The influence of platelet-rich plasma on myogenic differentiation. J Tissue Eng Regen Med 2016;10:E239-E249
  14. Zhao P, Jin Z, Chen Q, Yang T, Chen D, Meng J, Lu X, Gu Z, He Q. Local generation of hydrogen for enhanced photothermal therapy. Nat Commun 2018;9:4241
  15. Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, Katsura K, Katayama Y, Asoh S, Ohta S. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 2007;13:688-694
  16. Ono H, Nishijima Y, Adachi N, Sakamoto M, Kudo Y, Nakazawa J, Kaneko K, Nakao A. Hydrogen(H2) treatment for acute erythymatous skin diseases. A report of 4 patients with safety data and a non-controlled feasibility study with H2 concentration measurement on two volunteers. Med Gas Res 2012;2:14
  17. Guan P, Sun ZM, Luo LF, Zhou J, Yang S, Zhao YS, Yu FY, An JR, Wang N, Ji ES. Hydrogen protects against chronic intermittent hypoxia induced renal dysfunction by promoting autophagy and alleviating apoptosis. Life Sci 2019;225:46-54
  18. Li J, Hong Z, Liu H, Zhou J, Cui L, Yuan S, Chu X, Yu P. Hydrogen-rich saline promotes the recovery of renal function after ischemia/reperfusion injury in rats via anti-apoptosis and anti-inflammation. Front Pharmacol 2016;7:106
  19. Miyazaki N, Yamaguchi O, Nomiya M, Aikawa K, Kimura J. Preventive effect of hydrogen water on the development of detrusor overactivity in a rat model of bladder outlet obstruction. J Urol 2016;195:780-787
  20. Quan-Jun Y, Yan H, Yong-Long H, Li-Li W, Jie L, Jin-Lu H, Jin L, Peng-Guo C, Run G, Cheng G. Selumetinib attenuates skeletal muscle wasting in murine cachexia model through ERK inhibition and AKT activation. Mol Cancer Ther 2017;16:334-343
  21. Suzuki A, Minamide R, Iwata J. WNT/β-catenin signaling plays a crucial role in myoblast fusion through regulation of nephrin expression during development. Development 2018;145:dev168351
  22. Low S, Barnes JL, Zammit PS, Beauchamp JR. Delta-like 4 activates Notch 3 to regulate self-renewal in skeletal muscle stem cells. Stem Cells 2018;36:458-466
  23. Bohm A, Hoffmann C, Irmler M, Schneeweiss P, Schnauder G, Sailer C, Schmid V, Hudemann J, Machann J, Schick F, Beckers J, Hrabe de Angelis M, Staiger H, Fritsche A, Stefan N, Niess AM, Haring HU, Weigert C. TGF-β contributes to impaired exercise response by suppression of mitochondrial key regulators in skeletal muscle. Diabetes 2016;65:2849-2861
  24. Yi C, Liu D, Fong CC, Zhang J, Yang M. Gold nanoparticles promote osteogenic differentiation of mesenchymal stem cells through p38 MAPK pathway. ACS Nano 2010;4:6439-6448
  25. Lassar AB. The p38 MAPK family, a pushmi-pullyu of skeletal muscle differentiation. J Cell Biol 2009;187:941-943
  26. Jones NC, Tyner KJ, Nibarger L, Stanley HM, Cornelison DD, Fedorov YV, Olwin BB. The p38alpha/beta MAPK functions as a molecular switch to activate the quiescent satellite cell. J Cell Biol 2005;169:105-116
  27. Wakitani S, Saito T, Caplan AI. Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 1995;18:1417-1426
  28. Yang M, Dong Y, He Q, Zhu P, Zhuang Q, Shen J, Zhang X, Zhao M. Hydrogen: a novel option in human disease treatment. Oxid Med Cell Longev 2020;2020:8384742
  29. Ohta S. Molecular hydrogen as a preventive and therapeutic medical gas: initiation, development and potential of hydrogen medicine. Pharmacol Ther 2014;144:1-11
  30. Huang CS, Kawamura T, Toyoda Y, Nakao A. Recent advances in hydrogen research as a therapeutic medical gas. Free Radic Res 2010;44:971-982
  31. Smith GCS, Bouwmeester TM, Lam PH. Knotless double- row SutureBridge rotator cuff repairs have improved self-reinforcement compared with double-row SutureBridge repairs with tied medial knots: a biomechanical study using an ovine model. J Shoulder Elbow Surg 2017;26:2206-2212
  32. Kim KC, Shin HD, Cha SM, Park JY. Comparisons of retear patterns for 3 arthroscopic rotator cuff repair methods. Am J Sports Med 2014;42:558-565
  33. Killian ML, Cavinatto LM, Ward SR, Havlioglu N, Thomopoulos S, Galatz LM. Chronic degeneration leads to poor healing of repaired massive rotator cuff tears in rats. Am J Sports Med 2015;43:2401-2410
  34. Papa L, Djedaini M, Hoffman R. Mitochondrial role in stemness and differentiation of hematopoietic stem cells. Stem Cells Int 2019;2019:4067162
  35. Zhang H, Menzies KJ, Auwerx J. The role of mitochondria in stem cell fate and aging. Development 2018;145:dev143420
  36. Ciciliot S, Schiaffino S. Regeneration of mammalian skeletal muscle. Basic mechanisms and clinical implications. Curr Pharm Des 2010;16:906-914 Erratum in: Curr Pharm Des 2015;21:4657
  37. Yu X, Zhang Y, Li T, Ma Z, Jia H, Chen Q, Zhao Y, Zhai L, Zhong R, Li C, Zou X, Meng J, Chen AK, Puri PL, Chen M, Zhu D. Long non-coding RNA Linc-RAM enhances myogenic differentiation by interacting with MyoD. Nat Commun 2017;8:14016
  38. Paulin D, Li Z. Desmin: a major intermediate filament protein essential for the structural integrity and function of muscle. Exp Cell Res 2004;301:1-7
  39. Singh K, Dilworth FJ. Differential modulation of cell cycle progression distinguishes members of the myogenic regulatory factor family of transcription factors. FEBS J 2013;280:3991-4003
  40. Yokoyama S, Asahara H. The myogenic transcriptional network. Cell Mol Life Sci 2011;68:1843-1849
  41. Le Moal E, Pialoux V, Juban G, Groussard C, Zouhal H, Chazaud B, Mounier R. Redox control of skeletal muscle regeneration. Antioxid Redox Signal 2017;27:276-310
  42. Ji LL, Kang C, Zhang Y. Exercise-induced hormesis and skeletal muscle health. Free Radic Biol Med 2016;98:113-122
  43. Mason SA, Morrison D, McConell GK, Wadley GD. Muscle redox signalling pathways in exercise. Role of antioxidants. Free Radic Biol Med 2016;98:29-45
  44. Gibbons MC, Singh A, Engler AJ, Ward SR. The role of mechanobiology in progression of rotator cuff muscle atrophy and degeneration. J Orthop Res 2018;36:546-556
  45. Oh JH, Chung SW, Kim SH, Chung JY, Kim JY. 2013 neer award: effect of the adipose-derived stem cell for the improvement of fatty degeneration and rotator cuff healing in rabbit model. J Shoulder Elbow Surg 2014;23:445-455
  46. Pas HIMFL, Moen MH, Haisma HJ, Winters M. No evidence for the use of stem cell therapy for tendon disorders: a systematic review. Br J Sports Med 2017;51:996-1002
  47. Osborne H, Anderson L, Burt P, Young M, Gerrard D. Australasian College of Sports Physicians-position statement: the place of mesenchymal stem/stromal cell therapies in sport and exercise medicine. Br J Sports Med 2016;50:1237-1244
  48. Saito T, Sadoshima J. Molecular mechanisms of mitochondrial autophagy/mitophagy in the heart. Circ Res 2015;116:1477-1490
  49. Murakami Y, Ito M, Ohsawa I. Molecular hydrogen protects against oxidative stress-induced SH-SY5Y neuroblastoma cell death through the process of mitohormesis. PLoS One 2017;12:e0176992
  50. Qi R, Liu H, Wang Q, Wang J, Yang F, Long D, Huang J. Expressions and regulatory effects of P38/ERK/JNK Mapks in the adipogenic trans-differentiation of C2C12 myoblasts. Cell Physiol Biochem 2017;44:2467-2475
  51. Bernet JD, Doles JD, Hall JK, Kelly Tanaka K, Carter TA, Olwin BB. p38 MAPK signaling underlies a cell-autonomous loss of stem cell self-renewal in skeletal muscle of aged mice. Nat Med 2014;20:265-271
  52. Brien P, Pugazhendhi D, Woodhouse S, Oxley D, Pell JM. p38α MAPK regulates adult muscle stem cell fate by restricting progenitor proliferation during postnatal growth and repair. Stem Cells 2013;31:1597-1610
  53. Cuenda A, Sanz-Ezquerro JJ. p38γ and p38δ: from spectators to key physiological players. Trends Biochem Sci 2017;42:431-442