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ISSN 1527-3172 (Electronic)

2018, Number 3

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MEDICC Review 2018; 20 (3)

Heberprot-P’s effect on gene expression in healing diabetic foot ulcers

Camacho-Rodríguez H, Guillén-Pérez IA, Roca-Campaña J, Baldomero-Hernández JE, Tuero-Iglesias Á, Galván-Cabrera JA, Rodríguez-Cordero M, Palenzuela-Gardón DO, Berlanga-Acosta J, Novoa-Pérez LI

Language: English
References: 39
Page: 10-14
PDF size: 170.13 Kb.


ABSTRACT

INTRODUCTION Diabetic foot ulcers are a chronic complication in patients with diabetes mellitus. They appear as a result of the combination of diabetic polyneuropathy and angiopathy, and in many cases require amputation of the affected extremity. Clinical trials have demonstrated that repeated local infi ltration with Heberprot-P can improve healing of chronic diabetic foot ulcers. Although there is evidence of its effects as a granulation stimulator and on cell migration and proliferation, genetic control mechanisms explaining its anti-infl ammatory and oxidative stress reduction properties are not yet thoroughly understood.
OBJECTIVE Analyze changes in expression of genes involved in healing after treatment of diabetic foot ulcers with Heberprot-P.
METHODS Biopsies were collected from diabetic foot ulcers of 10 responding patients before and after 2 weeks’ treatment with Heberprot-P (75-μg applied intralesionally 3 times per week). Total RNA was obtained and quantitative PCR used to determine expression of 26 genes related to infl ammation, oxidative stress, cell proliferation, angiogenesis and extracellular matrix formation. Genetic expression was quantifi ed before and after treatment using REST 2009 v2.0.13.
RESULTS After treatment, there was a statistically signifi cant increase in expression of genes related to cell proliferation, angiogenesis and formation of extracellular matrix (PDGFB, CDK4, P21, TP53, ANGPT1, COL1A1, MMP2 and TIMP2). A signifi cant decrease was observed in gene expression related to infl ammatory processes and oxidative stress (NFKB1, TNFA and IL-1A).
CONCLUSIONS Our findings suggest that Heberprot-P’s healing action on diabetic foot ulcers is mediated through changes in genetic expression that reduce hypoxia, infl ammation and oxidative stress, and at the same time increase cell proliferation, collagen synthesis and extracellular matrix remodeling. The kinetics of expression of two genes related to extracellular matrix formation needs further exploration.


REFERENCES

  1. World Health Organization. Global Report on Diabetes [Internet]. Geneva: World Health Organization; 2016 [cited 2017 Jan 25]. 86 p. Available from: http://apps.who.int/iris/bitstream/ handle/10665/204871/9789241565257_eng.pdf; jsessionid=6D909E47C95E30D51430F445A8F7 592E?sequence=1

  2. Clayton W Jr, Elasy TA. A review of the pathophysiology, classifi cation, and treatment of foot ulcers in diabetic patients. Clin Diabetes. 2009 Apr;27(2):52–8.

  3. Reiber GE, Vileikyte L, Boyko EJ, del Aguila M, Smith DG, Lavery LA, et al. Causal pathways for incident lower extremity ulcers in patients with diabetes from two settings. Diabetes Care. 1999 Jan;22(1):157–62.

  4. National Health Statistics and Medical Records Division (CU). Anuario Estadístico de Salud 2016. Havana: Ministry of Public Health (CU); 2017 Apr. 206 p. Spanish.

  5. Giacco F, Brownlee M. Oxidative stress and diabetic complications. Circ Res. 2010 Oct 29;107(9):1058–70.

  6. Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature. 2001 Dec 13;414(6865):813–20.

  7. Bermúdez V, Bermúdez F, Arraiz N, Leal E, Linares S, Mengua E, et al. Biología molecular de los transportadores de glucosa: clasificación, estructura y distribución. Arch Venezolanos Farmacol Terap. 2007;26(2):76– 85. Spanish.

  8. Papanas N, Maltezos E. Etiology, pathophysiology and classifi cations of the diabetic Charcot foot. Diabet Foot Ankle. 2013 May 21;4. DOI: 10.3402/dfa.v4i0.20872.

  9. Forbes JM, Cooper ME. Mechanism of diabetic complication. Physiol Rev. 2013 Jan;93(1):137–88.

  10. Flanagan M. The physiology of wound healing. J Wound Care. 2000 Jun;9(6):299–300.

  11. Berlanga J, Fernández JI, López E, López PA, del Río A, Valenzuela C, et al. Heberprot-P: a novel product for treating advanced diabetic foot ulcer. MEDICC Rev. 2013 Jan;15(1):11–5.

  12. Zhao R, Liang H, Clarke E, Jackson C, Xue M. Infl ammation in chronic wounds. Int J Mol Sci. 2016 Dec 11;17(12). pii: E2085.

  13. Traversa B, Sussman G. The role of growth factors, cytokines and proteases in wound management. Growth factors, cytokines & proteases. 2001 Nov;9(4):161–7.

  14. Lodish H, Berk A, Matsudaira P, Kaiser C. Molecular Cell Biology. 5th ed. New York: W. H. Freeman; 2003 Aug 1. 973 p.

  15. Berg JM, Tymoczko JL, Stryer L. Biochemistry. 5th ed. New York: W. H. Freeman; 2002 Feb 15. 1100 p.

  16. Pfaffl MW. A new mathematical model for relative quantifi cation in real-time RT–PCR. Nucleic Acids Res. 2001 May 1;29(9):e45.

  17. Fernández-Montequín JI, Valenzuela-Silva CM, González-Díaz O, Savigne W, Sancho-Soutelo N, Rivero-Fernández F, et al. Intra-lesional injections of recombinant human epidermal growth factor promotes granulation and healing in advanced diabetic foot ulcers. Multicenter, randomized, placebo-controlled, double blind study. Int Wound J. 2009 Dec;6(6):432–43.

  18. Fleige S, Pfaffl MW. RNA integrity and the effect on the real-time qRT-PCR performance. Mol Aspects Med. 2006 Apr–Jun;27(2–3):126–39.

  19. Multid Analyses AB [Internet]. Göteborg (SE): MultiD Analyses AB; c2001–2015. GenEX 6.1 software; 2016 Feb 29 [cited 2017 May 27]. Available from: http://www.multid.se

  20. National Center for Biotechnology Information [Internet]. Bethesda (US): U.S. National Library of Medicine. Available from: https://www.ncbi .nlm.nih.gov/

  21. Rozen S, Skaletsky HJ. Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 2000;132:365–86.

  22. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002 Jun 18;3(7):RESEARCH0034. Epub 2002 Jun 18.

  23. Pfaffl MW, Horgan GW, Dempfl e L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res. 2002 May 1;30(9):e36.

  24. World Medical Association. Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA [Internet]. 2013 Nov 27 [cited 2018 May 5];310(20):2191–4. Available from: https://jamanetwork.com/journals/jama/full article/10.1001/jama.2013.281053

  25. Brownlee M. The pathobiology of diabetic complications: a unifying mechanism. Diabetes. 2005 Jun;54(6):1615–25.

  26. Goldin A, Beckman JA, Schmidt AM, Creager MA. Advanced glycation end products: sparking the development of diabetic vascular injury. Circulation. 2006 Aug 8;114(6):597–605.

  27. Noh H, King GL. The role of protein kinase C activation in diabetic nephropathy. Kidney Int Suppl. 2007 Aug;(106):S49–S53.

  28. Pierce GF, Mustoe TA, Altrock BW, Deuel TF, Thomason A. Role of platelet-derived growth factor in wound healing. J Cell Biochem. 1991 Apr;45(4):319–26. DOI:10.1002./jcb.240450403.

  29. Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev. 2003 Jul;83(3):835–70.

  30. Wang S, Nath N, Fusaro G, Chellappan S. Rb and prohibitin target distinct regions of E2F1 for repression and respond to different upstream signals. Mol Cell Biol. 1999 Nov;19(11):7447–60.

  31. Mishra S, Murphy LC, Murphy LJ. The Prohibitins: emerging roles in diverse functions. J Cellular Molecular Med. 2006;10(2):353–63.

  32. Ferry RJ Jr, Katz LE, Grimberg A, Cohen P, Weinzimer SA. Cellular actions of insulin-like growth factor binding proteins. Horm Metab Res. 1999 Feb–Mar;31(2–3):192–202.

  33. Liu Y, Min D, Bolton T, Nubé V, Twigg SM, Yue DK, et al. Increased matrix metalloproteinase-9 predicts poor wound healing in diabetic foot ulcers. Diabetes Care. 2009 Jan;32(1):117–9.

  34. Bhupinder S. Matrix metalloproteinase: an overview. Res Reports Biol. 2010 Sep 14;1:1–20.

  35. Armstrong DG, Jude EB. The role of matrix metalloproteases in wound healing. J Am Podiatric Med Assoc. 2002 Jan;92(1):12–8.

  36. Frazier K, Williams S, Kothapalli D, Klapper H, Grotendorst GR. Stimulation of fi broblast cell growth, matrix production, and granulation tissue formation by connective tissue growth factor. J Invest Dermatol. 1996 Sep;107(3):404–11.

  37. Pakyari M, Farrokhi A, Maharlooei MK, Ghahary A. Critical role of transforming growth factor beta in different phases of wound healing. Adv Wound Care (New Rochelle). 2013 Jun;2(5):215–24.

  38. Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev. 2003 Jul;83(3):835–70.

  39. Deonarine K, Panelli MC, Stashower ME, Jin P, Smith K, Slade HB, et al. Gene expression profi ling of cutaneous wound healing. J Transl Med. 2007 Feb 21;5:11.




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