Functional, morphological, and biochemical effects of Moringa oleifera leaf extract in mdx mice
Main Article Content
Abstract
Introduction: Duchenne muscular dystrophy (DMD) is the most common hereditary myopathy in childhood. The dystrophin protein, encoded by the DMD gene, is essential for maintaining the integrity of the muscle fiber sarcolemma, and its absence leads to progressive muscle degeneration. The lack of a known cure underscores the importance of alternative therapies aimed at mitigating disease progression. Moringa oleifera (MO) has been studied for its anti-inflammatory and antioxidant properties, which may contribute to reduced inflammation and muscle degeneration. Objective: To evaluate the effects of MO leaf extract on functional, morphological, and biochemical parameters in mdx mice, a murine model of DMD. Methods: Male mdx mice and C57BL/10 controls, aged 2 months, were treated for 8 weeks with MO leaf extract (300 mg/kg/day, oral gavage) or saline. Muscle strength was assessed using the inverted screen test. Morphological analysis of the quadriceps, extensor digitorum longus, tibialis anterior, and diaphragm included measurement of minimum Feret diameter and percentage of fibers with internalized nuclei. Serum levels of creatine kinase, AST/TGO, ALT/TGP, and urea were evaluated. Results: Treated dystrophic animals showed reduced AST/TGO levels, increased muscle strength, and a higher proportion of fibers with internalized nuclei, indicating modulation of regeneration and reduced tissue damage. Decreased AST/TGO suggests reduced sarcolemmal disruption, while increased internalized nuclei is consistent with an active regenerative response. Conclusion: MO treatment promoted functional improvement and partial attenuation of tissue damage, suggesting a beneficial role in regeneration and reduced muscle injury in dystrophic muscle.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (CC BY) that allows others to share and adapt the work with an acknowledgement of the work's authorship and initial publication in this journal.
Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.References
1. Crisafulli S, Sultana J, Fontana A, Salvo F, Messina S, Trifirò G. Global epidemiology of Duchenne muscular dystrophy: an updated systematic review and meta-analysis. Orphanet J Rare Dis. 2020;15(1):141. https://doi.org/10.1186/s13023-020-01430-8
2. Duan D, Goemans N, Takeda S, Mercuri E, Aartsma-Rus A. Duchenne muscular dystrophy. Nat Rev Dis Primers. 2021;7(1):13. https://doi.org/10.1038/s41572-021-00248-3
3. Mercuri E, Muntoni F. Muscular dystrophies. Lancet. 2013;381(9869):845-60. https://doi.org/10.1016/S0140-6736(12)61897-2
4. Darras BT, Menache-Starobinski CC, Hinton V, Kunkel LM. Dystrophinopathies. In: Darras BT, Jones H, Ryan M, Vivo DC. Neuromuscular Disorders of Infancy, Childhood, and Adolescence: a Clinician’s Approach. 2nd ed. London: Elsevier, 2015; p. 551-92.
5. Annexstad EJ, Lund-Petersen I, Rasmussen M. Duchenne muscular dystrophy. Tidsskr Nor Laegeforen. 2014;134(14):1361-4. https://doi.org/10.4045/tidsskr.13.0836
6. Passamano L, Taglia A, Palladino A, Viggiano E, D’Ambrosio P, Scutifero M, et al. Improvement of survival in Duchenne muscular dystrophy: retrospective analysis of 835 patients. Acta Myol. 2012;31(2):121-5.
7. Van Ruiten H, Bushby K, Guglieri M. State-of-the-art advances in Duchenne muscular dystrophy. Eur Med J. 2017;90-9.
8. United States Food and Drug Administration. FDA approves first gene therapy for treatment of certain patients with Duchenne Muscular Dystrophy. Available from: https://www.fda.gov/news-events/press-announcements/fda-approves-first-gene-therapy-treatment-certain-patients-duchenne-muscular-dystrophy.
9. Ng R, Banks GB, Hall JK, Muir LA, Ramos JN, Wicki J, et al. Animal models of muscular dystrophy. Prog Mol Biol Transl Sci. 2012;105:83-111. https://doi.org/10.1016/B978-0-12-394596-9.00004-4
10. Manning J, O’Malley D. What has the mdx mouse model of Duchenne muscular dystrophy contributed to our understanding of this disease? J Muscle Res Cell Motil. 2015;36(2):155-67. https://doi.org/10.1007/s10974-015-9406-4
11. Lessa TB, Dilayla KA, Bertassoli BM, Ambrosio CE. Arquitetura comparativa dos pulmões de camundongos normais e afetados pela distrofia muscular de Duchenne. Pesq Vet Bras. 2015;35(Suppl 1):56-60. https://doi.org/10.1590/S0100-736X2015001300010
12. Hoffman EP, Brown RH Jr, Kunkel LM. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell. 1987;51(6):919-28. https://doi.org/10.1016/0092-8674(87)90579-4
13. Zhou L, Rafael-Fortney JA, Huang P, Zhao XS, Cheng G, Zhou X, et al. Haploinsufficiency of utrophin gene worsens skeletal muscle inflammation and fibrosis in mdx mice. J Neurol Sci. 2008;264(1-2):106-11. https://doi.org/10.1016/j.jns.2007.08.029
14. Kou X, Li B, Olayanju JB, Drake JM, Chen N. Nutraceutical or pharmacological potential of Moringa oleifera Lam. Nutrients. 2018;10(3):343. https://doi.org/10.3390/nu10030343
15. Peterson JM, Kline W, Canan BD, Ricca DJ, Kaspar B, Delfín DA, et al. Peptide-based inhibition of NF-kappaB rescues diaphragm muscle contractile dysfunction in a murine model of Duchenne muscular dystrophy. Mol Med. 2011;17(5-6):508-15. https://doi.org/10.2119/molmed.2010.00263
16. Yang Q, Tang Y, Imbrogno K, Lu A, Proto JD, Chen A, et al. AAV-based shRNA silencing of NF-kappaB ameliorates muscle pathologies in mdx mice. Gene Ther. 2012;19(12):1196-204. https://doi.org/10.1038/gt.2011.207
17. Kumar A, Boriek AM. Mechanical stress activates the nuclear factor-kappaB pathway in skeletal muscle fibers: a possible role in Duchenne muscular dystrophy. FASEB J. 2003;17(3):386-96. https://doi.org/10.1096/fj.02-0542com
18. Hayden MS, Ghosh S. Regulation of NF-κB by TNF family cytokines. Semin Immunol. 2014;26(3):253-66. https://doi.org/10.1016/j.smim.2014.05.004
19. Nova E, Redondo-Useros N, Martínez-García RM, Gómez-Martínez S, Díaz-Prieto LE, Marcos A. Potential of Moringa oleifera to improve glucose control for the prevention of diabetes and related metabolic alterations: a systematic review of animal and human studies. Nutrients. 2020;12(7):2050. https://doi.org/10.3390/nu12072050
20. Ikezawa M, Minami N, Takahashi M, Goto Y, Miike T, Nonaka I. Dystrophin gene analysis on 130 patients with Duchenne muscular dystrophy with a special reference to muscle mRNA analysis. Brain Dev. 1998;20(3):165-8. https://doi.org/10.1016/S0387-7604(98)00012-6
21. Brookes PS, Yoon Y, Robotham JL, Anders MW, Sheu SS. Calcium, ATP, and ROS: a mitochondrial love-hate triangle. Am J Physiol Cell Physiol. 2004;287(4):C817-33. https://doi.org/10.1152/ajpcell.00139.2004
22. Zwart LL, Meerman JH, Commandeur JN, Vermeulen NP. Biomarkers of free radical damage applications in experimental animals and in humans. Free Radic Biol Med. 1999;26(1-2):202-26. https://doi.org/10.1016/s0891-5849(98)00196-8
23. Vasconcelos SML, Goulart MOF, Moura JBF, Manfredini V, Benfato LT, Kubota LT. Espécies reativas de oxigênio e de nitrogênio, antioxidantes e marcadores de dano oxidativo em sangue humano: principais métodos analíticos para sua determinação. Quím Nova. 2007;30(5):1323-38. https://doi.org/10.1590/S0100-40422007000500046
24. Duranti G, Maldini M, Crognale D, Horner K, Dimauro I, Sabatini S, et al. Moringa oleifera leaf extract upregulates Nrf2/HO-1 expression and ameliorates redox status in C2C12 skeletal muscle cells. Molecules. 2021;26(16):5041. https://doi.org/10.3390/molecules26165041
25. Sreelatha S, Padma PR. Protective mechanisms of Moringa oleifera against CCl4-induced oxidative stress in precision-cut liver slices. Forsche Komplementmed. 2010;17(4):189-94. https://doi.org/10.1159/000318606
26. Pareek A, Pant M, Gupta MM, Kashania P, Ratan Y, Jain V, et al. Moringa oleifera: an updated comprehensive review of its pharmacological activities, ethnomedicinal, phytopharmaceutical formulation, clinical, phytochemical, and toxicological aspects. Int J Mol Sci. 2023;24(3):2098. https://doi.org/10.3390/ijms24032098
27. Grupo Rocha Saúde. Chá de moringa. Available from: https://www.gruporochasaude.com/chas/cha-de-moringa.
28. Perazzo FF, Carvalho JCT, Carvalho JE, Rehder VLG. Central properties of the essential oil and the crude ethanol extract from aerial parts of Artemisia annua L. Pharmacol Res. 2003;48:497-502. https://doi.org/10.1016/s1043-6618(03)00216-0
29. van Putten M, Aartsma-Rus A, Dorchies O, Nagaraju K, Carlson G. The use of hanging wire tests to monitor muscle strength and condition over time. Available from: https://www.treat-nmd.org/wp-content/uploads/2023/07/DMD_M_2.1.004.pdf.
30. Folker ES, Baylies MK. Nuclear positioning in muscle development and disease. Front Physiol. 2013;4:363. https://doi.org/10.3389/fphys.2013.00363
31. Lazzarin MC. Caracterização da regeneração, inflamação e estresse oxidativo no músculo de modelo experimental para distrofia muscular de Duchenne com fenótipo exacerbado pelo exercício [dissertation]. São Paulo: Universidade Federal de São Paulo; 2021.
32. Departamento de Biologia Estrutural e Funcional, Instituto de Biologia, Universidade Estadual de Campinas. Características histopatológicas e processo inflamatório do músculo quadríceps de camundongos mdx, modelo experimental da distrofia muscular de Duchenne. Campinas: UNICAMP; 2023.
33. van Putten M, Putker K, Overzier M, Adamzek WA, Pasteuning-Vuhman S, Plomp JJ, et al. Natural disease history of the D2-mdx mouse model for Duchenne muscular dystrophy. FASEB J. 2019;33(7):8110-24. https://doi.org/10.1096/fj.201802488R
34. Vergara-Jimenez M, Almatrafi MM, Fernandez ML. Bioactive components in Moringa oleifera leaves protect against chronic disease. Antioxidants (Basel). 2017;6(4):91. https://doi.org/10.3390/antiox6040091
35. Donovan J, Phan H, Russell A, Barthel B, Thaler L, Kilburn N, et al. 351P Sevasemten, a fast myosin inhibitor, in adults with Becker muscular dystrophy results in reduced muscle damage biomarkers and functional stabilization. Neuromuscular Disord. 2024;43(Suppl 1):104441. https://doi.org/10.1016/j.nmd.2024.07.691
36. Barodia K, Cheruku SP, Kanwal A, Menon A, Rajeevan R, Rukade A, et al. Effect of Moringa oleifera leaf extract on exercise and dexamethasone-induced functional impairment in skeletal muscles. J Ayurveda Integr Med. 2022;13(1):100503. https://doi.org/10.1016/j.jaim.2021.07.019
37. Grounds MD, White JD, Rosenthal N, Korneluk R. The role of stem cells in skeletal and cardiac muscle repair. J Histochem Cytochem. 2008;56(9):891-901. https://doi.org/10.1177/002215540205000501
38. Chargé SBP, Rudnicki MA. Cellular and molecular regulation of muscle regeneration. Physiol Rev. 2004;84(1):209-38. https://doi.org/10.1152/physrev.00019.2003
39. Dubowitz V, Sewry CA, Oldfors A. Muscle Biopsy: A Practical Approach. 4th ed. Philadelphia: Saunders Elsevier; 2013.
40. Guiraud S, Davies K. Regenerative biomarkers for Duchenne muscular dystrophy. Neural Regen Res. 2019;14(8):1317-20. https://doi.org/10.4103/1673-5374.253534