Aloe vera stimulate cell proliferation, cell migration, expression of vascular endothelial growth factor-A (VEGF-A), and c-Jun N-terminal kinase-1 (JNK-1) on fibroblast of diabetic rat models

https://doi.org/10.19106/JMedSci005102201903

Insanul Firdaus(1*), Nur Arfian(2), Mae Sri Hartati Wahyuningsih(3), Denny Agustiningsih(4)

(1) Basic Medical & Biomedical Science, Faculty of Medicine, Universitas Gadjah Mada
(2) Departement of Anatomy and Embryology, Faculty of Medicine Universitas Gadjah Mada
(3) Departement of Pharmacology and Therapy, Faculty of Medicine Universitas Gadjah Mada
(4) Departement of Physiology, Faculty of Medicine Universitas Gadjah Mada
(*) Corresponding Author

Abstract


The disturbance of cell migration and cell proliferation,diminished production of vascular endothelial growth factor-A (VEGF-A) and c-Jun N-terminal kinase-1 (JNK-1) are important factors in wound healing process. Aloe vera contains active compounds which can help in the wound healing process. Thestudy aimed to investigate the effect of ethanol extract ofA. veraon cell proliferation, cell migration, VEGF-A and JNK-1 expression of skin fibroblast cells of diabetic rats. The primary skin fibroblast cells were isolated from diabetic Wistar rat and incubated with the A. vera extract in various concentrations i.e. 500 (AV500), 250 (AV250), and 125 µg/Ml (AV125) for 24, 48 and 72 h.The cell proliferation was examined visually by counting the cells number, the cell migration was observed using in vitro scratch assay, whereas VEGF-A and JNK-1 expression were examined using RT-PCR. In 24 and 48h incubation,the cell proliferation of AV500 and AV250 groups had higher number of cells than negative control group,but there was no significant difference (p>0.05). However in72 h incubation,the cell proliferation of AV500 group (29.33±1.28x104 cells/mL)was significantly different compared to negative control group (22.91±3.21x104 cells/mL) (p<0.05). In 24 h incubation, the cell migration of AV500(78.13±7.18%), AV250 (73.88±4.75%) and AV125 (68.80±17.11%) groupswere significantly higher thanthat of negative control group (53.91±2.74%) (p<0.05). In contrastin 48 and 72 hincubation,there were no significantly different in cell migration (p>0.05).The expression of VEGF-A and JNK-1 after incubation with the AV500 for 48 h, weresignificantly higher than those of negative control group (p<0.05).In conclusion, A. vera increases cell proliferation, cell migration, VEGF-A and JNK-1 expression offibroblast cellof diabetic rat skin.


Keywords


Diabetic ulcer; Aloe vera; fibroblast cell culture; cell proliferation; cell migration; VEGF-A expression; JNK-1 expression

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References

  1. Alberti KG, Zimmet PZ. Definition, diagnosis and classification of diabetes mellitus and its complications. Part 1: diagnosis and classification of diabetes mellitus. Provisional report of a WHO consultation. Diabet Med. 1998; 15(7):539-53. https://doi.org/10.1002/(SICI)1096-9136(199807)15:7<539::AID-DIA668>3.0.CO;2-S
  2. Noor S, Zubair M, Ahmad J. Diabetic foot ulcer- a review on pathophysiology, classification and microbial etiology. Diabetes Metab Syndr Clin Res Rev 2015; 9(3):192-9. https://doi.org/10.1016/j.dsx.2015.04.007
  3. Blakytny R, Jude E. The molecular biology of chronic wounds and delayed healing in diabetes. Diabet Med 2006; 23(6):594-608. https://doi.org/10.1111/j.1464-5491.2006.01773.x
  4. Chithra P, Sajithlal GB, Chandrakasan G. Influence of Aloe vera on the healing of dermal wounds in diabetic rats. J Ethnopharmacol. 1998; 59(3):195-201. https://doi.org/10.1016/S0378-8741(97)00124-4
  5. Lien Y-H, Tseng MM, Stern R. Glucose and glucose analogs modulate collagen metabolism. Exp Mol Pathol 1992;57(3):215-21. https://doi.org/10.1016/0014-4800(92)90012-Z
  6. Folkman J. Angiogenesis inhibitors generated by tumors. Mol Med 1995; 1(2):120. https://doi.org/10.1007/BF03401559
  7. Lamers ML, Almeida MES, Vicente-Manzanares M, Horwitz AF, Santos MF. High glucose-mediated oxidative stress impairs cell migration. Blagosklonny MV. PLoS One. 2011; 6(8):e22865. https://doi.org/10.1371/journal.pone.0022865
  8. Lerman OZ, Galiano RD, Armour M, Levine JP, Gurtner GC. Cellular dysfunction in the diabetic fibroblast: impairment in migration, vascular endothelial growth factor production, and response to hypoxia. Am J Pathol. 2003; 162(1):303-12. https://doi.org/10.1016/S0002-9440(10)63821-7
  9. Ferrara N. Role of vascular endothelial growth factor in regulation of physiological angiogenesis. Am J Physiol-Cell Physiol. 2001; 280(6):C1358-66. https://doi.org/10.1152/ajpcell.2001.280.6.C1358
  10. Johnson KE, Wilgus TA. Vascular endothelial growth factor and angiogenesis in the regulation of cutaneous wound repair. Adv Wound Care 2014; 3(10):647-61. https://doi.org/10.1089/wound.2013.0517
  11. Hehenberger K, Heilborn JD, Brismar K, Hansson A. Inhibited proliferation of fibroblasts derived from chronic diabetic wounds and normal dermal fibroblasts treated with high glucose is associated with increased formation of L-lactate. Wound Repair Regen 1998; 6(2):135-41. https://doi.org/10.1046/j.1524-475X.1998.60207.x
  12. Desta T, Li J, Chino T, Graves DT. Altered fibroblast proliferation and apoptosis in diabetic gingival wounds. J Dent Res 2010; 89(6):609-14. https://doi.org/10.1177/0022034510362960
  13. Xuan YH, Huang BB, Tian HS, Chi LS, Duan YM, Wang X, et al. High-glucose inhibits human fibroblast cell migration in wound healing via repression of bFGF-regulating JNK phosphorylation. PLoS One 2014; 9(9):e108182. https://doi.org/10.1371/journal.pone.0108182
  14. Rajeswari R, Umadevi M, Rahale CS, Selvavenkadesh S, Kumar KS, Bhowmik D. Aloe vera: the miracle plant its medicinal and traditional uses in India. J Pharmacogn Phytochem 2012; 1(4): 118-24.
  15. Dat AD, Poon F, Pham KB, Doust J. Aloe vera for treating acute and chronic wounds. In: The Cochrane Collaboration, editor. Cochrane Database of Systematic Reviews. Chichester, UK: John Wiley & Sons, Ltd; 2012. https://doi.org/10.1002/14651858.CD008762.pub2
  16. Takzare N, Hosseini M, Hasanzadeh G, Mortazavi H, Takzare A, Habibi P. Influence of Aloe vera gel on dermal wound healing process in rat. Toxicol Mech Methods 2009; 19(1):73-7. https://doi.org/10.1080/15376510802442444
  17. Yao H, Chen Y, Li S, Huang L, Chen W, Lin X. Promotion proliferation effect of a polysaccharide from Aloe barbadensis Miller on human fibroblasts in vitro. Int J Biol Macromol 2009; 45(2):152-6. https://doi.org/10.1016/j.ijbiomac.2009.04.013
  18. Curto EM, Labelle A, Chandler HL. Aloe vera: an in vitro study of effects on corneal wound closure and collagenase activity. Vet Ophthalmol. 2014; 17(6):403-410. https://doi.org/10.1111/vop.12163
  19. Moon E-J, Lee YM, Lee O-H, Lee M-J, Lee S-K, Chung M-H, et al. A novel angiogenic factor derived from Aloe vera gel: β-sitosterol, a plant sterol. Angiogenesis 1999; 3(2):117-23. https://doi.org/10.1023/A:1009058232389
  20. Jettanacheawchankit S, Sasithanasate S, Sangvanich P, Banlunara W, Thunyakitpisal P. Acemannan stimulates gingival fibroblast proliferation; expressions of keratinocyte growth factor-1, vascular endothelial growth factor, and type I collagen; and wound healing. J Pharmacol Sci 2009; 109(4):525-31. https://doi.org/10.1254/jphs.08204FP
  21. Atiba A, Ueno H, Uzuka Y. The effect of Aloe vera oral administration on cutaneous wound healing in type 2 diabetic rats. J Vet Med Sci. 2011; 73(5):583-9. https://doi.org/10.1292/jvms.10-0438
  22. Moniruzzaman M, Rokeya B, Ahmed S, Bhowmik A, Khalil M, Gan S. In vitro antioxidant effects of Aloe barbadensis Miller extracts and the potential role of these extracts as antidiabetic and antilipidemic agents on streptozotocin-induced type 2 diabetic model rats. Molecules 2012; 17(12):12851-67. https://doi.org/10.3390/molecules171112851
  23. Szkudelski T. Streptozotocin – nicotinamide - induced diabetes in the rat. Characteristics of the experimental model. Exp Biol Med 2012; 237(5):481-90. https://doi.org/10.1258/ebm.2012.011372
  24. Seluanov A, Vaidya A, Gorbunova V. Establishing primary adult fibroblast cultures from rodents. JOVE 2010; (44): pii:2033. https://doi.org/10.3791/2033
  25. Liu Z, Lei M, Jiang Y, Hao H, Chu L, Xu J, et al. High glucose attenuates VEGF expression in rat multipotent adult progenitor cells in association with inhibition of JAK2/STAT3 signalling. J Cell Mol Med 2009; 13(9b):3427-36. https://doi.org/10.1111/j.1582-4934.2008.00502.x
  26. Huang C. MAP kinases and cell migration. J Cell Sci 2004; 117(20):4619-28. https://doi.org/10.1242/jcs.01481
  27. Choi S, Kim K-W, Choi J-S, Han S-T, Park Y-I, Lee S-K, et al. Angiogenic activity of β-sitosterol in the ischaemia/reperfusion-damaged brain of Mongolian gerbil. Planta Med 2002; 68(4):330-5. https://doi.org/10.1055/s-2002-26750
  28. Atiba A, Nishimura M, Kakinuma S, Hiraoka T, Goryo M, Shimada Y, et al.Aloe vera oral administration accelerates acute radiation-delayed wound healing by stimulating transforming growth factor-β and fibroblast growth factor production. Am J Surg 2011; 201(6):809-18. https://doi.org/10.1016/j.amjsurg.2010.06.017
  29. Abdullah KM, Abdullah A, Johnson ML, Bilski JJ, Petry K, Redmer DA, et al. Effects of Aloe vera on gap junctional intercellular communication and proliferation of human diabetic and nondiabetic skin fibroblasts. J Altern Complement Med 2003; 9(5):711-8. https://doi.org/10.1089/107555303322524553
  30. Lee KY, Park JH, Chung MH, Park YI, Kim KW, Lee YJ, et al. Aloesin up-regulates cyclin E/CDK2 kinase activity via inducing the protein levels of cyclin E, CDK2, and CDC25A in SK-HEP-1 cells. IUBMB Life. 1997; 41(2):285-292. https://doi.org/10.1080/15216549700201291
  31. Boudreau MD, Beland FA. An evaluation of the biological and toxicological properties of Aloe barbadensis (Miller), Aloe vera. J Environ Sci Health Part C 2006; 24(1):103-154. https://doi.org/10.1080/10590500600614303
  32. Daburkar M, Rathore A, Tangadpaliwar S, Bhutada P, Lohar V. An in vivo and in vitro investigation of the effect of Aloe vera gel ethanolic extract using animal model with diabetic foot ulcer. J Pharm Bioallied Sci 2014; 6(3):205-12. https://doi.org/10.4103/0975-7406.135248
  33. Davis RH, Donato JJ, Hartman GM, Haas RC. Anti inflamatory and wound healing activity of a growth substance in Aloe vera. J Am Podiatr Med Assoc 1994;84(2):77-82. https://doi.org/10.7547/87507315-84-2-77
  34. Xing W, Guo W, Zou C-H, Fu T-T, Li X-Y, Zhu M, et al. Acemannan accelerates cell proliferation and skin wound healing through AKT/mTOR signaling pathway. J Dermatol Sci 2015; 79(2):101-9. https://doi.org/10.1016/j.jdermsci.2015.03.016
  35. Ozsoy N, Candoken E, Akev N. Implications for degenerative disorders: antioxidative activity, total phenols, flavonoids, ascorbic acid, β-carotene and α-tocopherol in Aloe vera. Oxid Med Cell Longev 2009; 2(2):99-106.https://doi.org/10.4161/oxim.2.2.8493



DOI: https://doi.org/10.19106/JMedSci005102201903

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