L-Carnitine Promotes Cardiomyogenic Differentiation of C-Kit+Bone Marrow Progenitor Cells via MAPK-ERK Signaling Pathway
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Abstract
Many studies have shown that bone marrow (BM) stem/progenitor cells have the highest probability of cardiomyocyte differentiation. Regarding the major role of C-kit+ BM stem cells in cell therapy of patients with heart disease and getting cells with higher differentiation potential, this study aimed to investigate the capacity and effect of L-carnitine (LC) on cardiomyogenic differentiation of C-kit+ BM cells through MAPK/ERK signaling pathway. For this purpose, C-kit+ was enriched from the BM mononuclear cell population using a magnetic activating cell sorting technique. The purity of the separated C-kit+ cells was then evaluated by flow cytometry. In the next step, C-Kit+ cells were treated in a cardiomyogenic differentiation culture medium for 21 days once in the presence and once in the absence of 0.2 µM LC (the experimental and control groups). To evaluate the cardiomyogenic differentiation potential of C-kit+ cells, the Desmin cell marker was determined by immunocytochemistry. The expressions of both GATA4 and ERK proteins were measured using western blotting and flow cytometry, respectively. The results show that 95.7 percent of the cells separated by the MACS technique expressed a C-kit+ cell marker. Additionally, it was found that 0.2 mM LC significantly increased the expression of GATA4 protein in the cardiomyogenic differentiated cells. The expression of ERK protein also suggested a significant increase of about 1.60 times in the experimental group in comparison with the control group (*P˂0.05). In brief, it was found that treating C-kit+ BM cells with LC increases cardiomyogenic differentiation by increasing the expression of GATA4. Notably, this effect can take place through MARK/ERK signaling pathway. The results of this research can be valuable in suggesting a treatment solution for cardiovascular diseases.
2. Mitra A, Basak, Datta TK, Naskar S, Sengupta S, Sarkar S. Role of α-crystallin B as a regulatory switch in modulating cardiomyocyte apoptosis by mitochondria or endoplasmic reticulum during cardiac hypertrophy and myocardial infarction. Cell Death Dis 2013;4:e582.
3. Shafei AE, Ali MA, Ghanem HG, Shehata AI, Abdelgawad AA, Handal HR, et al. Mesenchymal stem cell therapy: A promising cell‐based therapy for treatment of myocardial infarction. J Gene Med 2017;19:e2995.
4. Bahit MC, Kochar A, Granger CB, Post-myocardial infarction heart failure. JACC Heart Fail 2018;6:179-86.
5. Zaruba MM, Soonpaa M, Mark Soonpaa, Field LJ. Cardiomyogenic potential of c-kit+ expressing cells derived from neonatal and adult mouse hearts. Circulation 2010;121:1992-2000.
6. Gambini E, Pompilio G, Biondi A, Alamanni F, Capogrossi MC, Agrifoglio M, et al. C-kit+ cardiac progenitors exhibit mesenchymal markers and preferential cardiovascular commitment. Cardiovasc Res 2011;89:362-73.
7. Pekala J, Patkowska-Sokoła B, Bodkowski R, Jamroz D, Nowakowski P, Lochyński S, et al. L-carnitine-metabolic functions and meaning in humans life. Curr Drug Metab 2011;12:667-78.
8. Karlic H, Lohninger A. Supplementation of L-carnitine in athletes: does it make sense? Nutrition 2004;20:709-15.
9. Wang ZY, Liu YY, Liu GH, Lu HB, Mao CY. l-Carnitine and heart disease. Life Sci 2018;194:88-97.
10. Muslin AJ. MAPK signalling in cardiovascular health and disease: molecular mechanisms and therapeutic targets. Clin Sci (Lond) 2008;115:203-18.
11. Sun Y, Liu WW, Liu T, Feng X, Yang N, Zhou HF. Signaling pathway of MAPK/ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct Res 2015;35:600-4.
12. Hu B, Song JT, Ji XF, Liu ZQ, Cong ML, Liu DX. Sodium ferulate protects against angiotensin II-induced cardiac hypertrophy in mice by regulating the MAPK/ERK and JNK pathways. BioMed Res Int 2017;2017:3754942.
13. Hao T, Zhou J, Lü S, Yang B, Wang Y, Fang W, et al. Fullerene mediates proliferation and cardiomyogenic differentiation of adipose-derived stem cells via modulation of MAPK pathway and cardiac protein expression. Int J Nanomedicine 2016;11:269-83.
14. Liu P, Zhong TP. MAPK/ERK signalling is required for zebrafish cardiac regeneration. Biotechnol Lett 2017;39:1069-77.
15. Khaksar M, Sayyari M, Rezaie J, Pouyafar A, Montazersaheb S, Rahbarghazi R. High glucose condition limited the angiogenic/cardiogenic capacity of murine cardiac progenitor cells in in vitro and in vivo milieu. Cell Biochem Funct 2018;36:346-56.
16. Farahzadi R, Fathi E, Vietor I. Mesenchymal stem cells could be considered as a candidate for further studies in cell-based therapy of Alzheimer’s disease via targeting the signaling pathways. ACS Chem Neurosci 2020;11:1424-35.
17. Fathi E, Farahzadi R, Javanmardi S, Vietor I. L-carnitine Extends the Telomere Length of the Cardiac Differentiated CD117+-Expressing Stem Cells. Tissue Cell 2020;67:101429.
18. Tarhriz V, Wagner KD, Masoumi Z, Molavi O, Hejazi MS, Ghanbarian H. CDK9 regulates apoptosis of myoblast cells by modulation of microRNA‐1 expression. J Cell Biochem 2018;119:547-54.
19. Tarhriz V, Eyvazi S, Musavi M, Abasi M, Sharifi K, Ghanbarian H, et al. Transient induction of Cdk9 in the early stage of differentiation is critical for myogenesis. J Cell Biochem 2019;120:18854-61.
20. Mehdizadeh A, Somi MH, Darabi M, Farajnia S, Akbarzadeh A, Montazersaheb S, et al. Liposome-mediated RNA interference delivery against Erk1 and Erk2 does not equally promote chemosensitivity in human hepatocellular carcinoma cell line HepG2. Artif Cells Nanomed Biotechnol 2017;45:1612-9.
21. Fathi E, Valipour B, Farahzadi R. Targeting the Proliferation Inhibition of Chronic Myeloid Leukemia Cells by Bone Marrow Derived-Mesenchymal Stem Cells via ERK Pathway as a Therapeutic Strategy. Acta Med Iran 2020;58:199-206.
22. Loffredo FS, Steinhauser ML, Gannon J, Lee RT. Bone marrow-derived cell therapy stimulates endogenous cardiomyocyte progenitors and promotes cardiac repair. Cell Stem Cell 2011;8:389-98.
23. Lagostena L, Avitabile D, De Falco E, Orlandi A, Grassi F, Iachininoto MG, et al. Electrophysiological properties of mouse bone marrow c-kit+ cells co-cultured onto neonatal cardiac myocytes. Cardiovasc Res 2005;66:482-92.
24. Van Berlo JH, Kanisicak O, Maillet M, Vagnozzi RJ, Karch J, Lin SC, et al. C-kit+ cells minimally contribute cardiomyocytes to the heart. Nature 2014;509:337-41.
25. Matuszczak S, Czapla J, Jarosz-Biej M, Wiśniewska E, Cichoń T, Smolarczyk R, et al. Characteristic of c-Kit+ progenitor cells in explanted human hearts. Clin Res Cardiol 2014;103:711-8.
26. Fazel S, Cimini M, Chen L, Li S, Angoulvant D, Fedak P, et al. Cardioprotective c-kit+ cells are from the bone marrow and regulate the myocardial balance of angiogenic cytokines. J Clin Invest 2006;116:1865-77.
27. Xue C, Zhang J, Lv Z, Liu H, Huang C, Yang J, et al., Angiotensin II promotes differentiation of mouse c-kit-positive cardiac stem cells into pacemaker-like cells. Mol Med Rep 2015;11:3249-58.
28. Gojo S, Gojo N, Takeda Y, Mori T, Abe H, Kyo S, et al. In vivo cardiovasculogenesis by direct injection of isolated adult mesenchymal stem cells. Exp Cell Res 2003;288:51-9.
29. Kubo H, Berretta RM, Jaleel N, Angert D, Houser SR. c‐Kit+ Bone Marrow Stem Cells Differentiate into Functional Cardiac Myocytes. Clin Transl Sci 2009;2:26-32.
30. Martinnzzi A, Vergani L, Rosa M, Angelini C. L-Carnitine uptake in differentiating human cultured muscle. Biochim Biophys Acta 1991;1095:217-22.
31. Winter SC, Buist NR. Cardiomyopathy in childhood, mitochondrial dysfunction, and the role of L-carnitine. Am Heart J 2000;139:s63-9.
32. Mingorance C, Rodríguez-Rodríguez R, Justo ML, Alvarez de Sotomayor M, Herrera MD. Critical update for the clinical use of L-carnitine analogs in cardiometabolic disorders. Vasc Health Risk Manag 2011;7:169-76.
33. Fathi E, Farahzadi R, Vietor I, Javanmardi S. Cardiac differentiation of bone-marrow-resident c-kit+ stem cells by L-carnitine increases through secretion of VEGF, IL6, IGF-1, and TGF-β as clinical agents in cardiac regeneration. J Biosci 2020;45:92.
34. Kempf H, Lecina M, Ting S, Zweigerdt R, Oh S. Distinct regulation of mitogen-activated protein kinase activities is coupled with enhanced cardiac differentiation of human embryonic stem cells. Stem Cell Res 2011;7:198-209.
| Files | ||
| Issue | Vol 61 No 4 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/acta.v61i4.13172 | |
| Keywords | ||
| C-kit bone marrow progenitor cells Cardiomyogenic differentiation L-carnitin MAPK-ERK signaling pathway | ||
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