Azize ŞENER, Özge ÇEVİK, Özge DOĞAN, Nazli Gül ALTINDİŞ, Halil AKSOY, Betül OKUYAN

The effects of topical melatonin on oxidative stress, apoptosis signals,and p53 protein expression during cutaneous wound healing

Elimination of reactive oxygen species (ROS) can be an important strategy to improve healing of wounds. ROS have an effect on proliferation and cell survival signaling, which results in alteration of apoptotic pathways in cells. Melatonin has antioxidant properties on skin wounds. In our study, we investigated the effects of topical melatonin (3%, w/w) on apoptosis and p53 protein expression together with parameters of oxidative stress in a cutaneous excision wound model. Bcl-2 protein levels in wound tissue at the end of days 3, 7, and 14 were significantly increased, while caspase-3 activity and p53 protein expression in wound tissue at the end of days 3, 7, and 14 were also reduced with melatonin treatment during wound healing. On days 3 and 7 after the wound, malondialdehyde level was reduced and glutathione was increased with melatonin treatment. Melatonin decreased myeloperoxidase levels and increased hydroxyproline levels in wound tissue at the end of day 7. However, melatonin had no significant effect on percentage of wound closure. Considering our results, topical melatonin displays antioxidant, antiapoptotic, and p53-inhibitory effects, but these effects are not sufficient for the acceleration of wound closure.

Anahtar Kelimeler:

Apoptosis, Bcl-2, p53, oxidative stress, topical melatonin, wound healing

The effects of topical melatonin on oxidative stress, apoptosis signals,and p53 protein expression during cutaneous wound healing

Keywords:

Apoptosis, Bcl-2, p53, oxidative stress, topical melatonin, wound healing,

PDF

___

Baydas G, Reiter RJ, Akbulut M, Tuzcu M, Tamer S (2005). Melatonin inhibits neural apoptosis induced by homocysteine in hippocampus of rats via inhibition of cytochrome c translocation and caspase-3 activation and by regulating pro- and anti-apoptotic protein levels. Neuroscience 135: 879–886.
Beutler E (1975). Glutathione. In: Beutler E ,editor. Red Cell Metabolism, A Manual of Biochemical Methods. 2nd ed. New York, NY, USA: Grune and Stratton, pp. 112–114.
Blakytny R, Jude E (2006). The molecular biology of chronic wounds and delayed healing in diabetes. Diabet Med 23: 594–608.
Bradford MM (1976). Rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254.
Buege JA, Aust SD (1978). Microsomal lipid peroxidation. Methods Enzymol 52: 302–310.
Bulbuller N, Dogru O, Yekeler H, Cetinkaya Z, Ilhan N, Kirkil C (2005). Effect of melatonin on wound healing in normal and pinealectomized rats. J Surg Res 123: 3–7.
Chang WK, Yang KD, Chuang H, Jan JT, Shaio MF (2002). Glutamine protects activated human T cells from apoptosis by up-regulating glutathione and Bcl-2 levels. Clin Immunol 104: 151–160.
Cuzzocrea S, Tan DX, Costantino G, Mazzon E, Caputi AP, Reiter RJ (1999). The protective role of endogenous melatonin in carrageenan-induced pleurisy in the rat. FASEB J 13: 1930– 1938.
Deveci M, Gilmont RR, Dunham WR, Mudge BP, Smith DJ, Marcelo CL (2005). Glutathione enhances fibroblast collagen contraction and protects keratinocytes from apoptosis in hyperglycaemic culture. Br J Dermatol 152: 217–224.
Drobnik J (2012). Wound healing and the effect of pineal gland and melatonin. J Exp Integr Med 2: 3–14.
Drobnik J, Dabrowski R (1996). Melatonin influence on the collagen content in the granulation tissue of a wound depends on the time of its application. J Endocrinol Invest 19: 48–48.
Fan TJ, Han LH, Cong RS, Liang J (2005). Caspase family proteases and apoptosis. Acta Biochim Biophys Sin (Shanghai) 37: 719– 727.
Fischer TW, Slominski A, Zmijewski MA, Reiter RJ, Paus R (2008a). Melatonin as a major skin protectant: from free radical scavenging to DNA damage repair. Exp Dermatol 17: 713–730.
Fischer TW, Zmijewsk MA. Wortsman J, Slominski A (2008b). Melatonin maintains mitochondrial membrane potential and attenuates activation of initiator (casp-9) and effector caspases (casp-3/casp-7) and PARP in UVR-exposed HaCaT keratinocytes. J Pineal Res 44: 397–407.
Greenhalgh DG (1998). The role of apoptosis, wound healing. Int J Biochem Cell Biol 30: 1019–1030.
Hameedaldeen A, Liu J, Batres A, Graves GS, Graves DT (2014). FOXO1, TGF-β regulation and wound healing. Int J Mol Sci 15: 16257–16269.
Hillegas, LM, Griswold DE, Brickson B, Albrightson-Winslow C (1990). Assessment of myeloperoxidase activity in whole rat kidney. J Pharmacol Methods 24: 285–295.
James TJ, Hughes MA, Hofman D, Cherry GW, Taylor RP (2001). Antioxidant characteristics of chronic wound fluid. Br J Dermatol 145: 185–186.
Kane CD, Greenhalgh DG (2000). Expression and localization of p53 and Bcl-2 in healing wounds in diabetic and nondiabetic mice. Wound Repair Regen 8: 45–58.
Kim BC, Shon BS, Ryoo YW, Kim SP, Lee KS (2001). Melatonin reduces X-ray irradiation-induced oxidative damages in cultured human skin fibroblasts. J Dermatol Sci 26: 194–200.
Lippens S, Hoste E, Vandenabeele P, Agostinis P, Declercq W (2009). Cell death in the skin. Apoptosis 14: 549–569.
Nagata M, Takenaka H, Shibagaki R, Kishimoto S (1999). Apoptosis and p53 protein expression increase in the process of burn wound healing in guinea-pig skin. Br J Dermatol 140: 829–838.
Ozler M, Korkmaz A, Uysal B, Simsek K, Ozkan C, Topal T, Oter S (2011). Effects of topical melatonin and vitamin E in a rat ischemic wound model. J Exp Integr Med 1: 123–129.
Ozler M, Simsek K, Ozkan C, Akgul EO, Topal T, Oter S, Korkmaz A (2010). Comparison of the effect of topical and systemic melatonin administration on delayed wound healing in rats that underwent pinealectomy. Scand J Clin Lab Invest 70: 447–452.
Rai NK, Tripathi K, Sharma D, Shukla VK (2005). Apoptosis: a basic physiologic process in wound healing. Int J Low Extrem Wounds 4: 138–144.
Reddy GK, Enwemeka CS (1996). A simplified method for the analysis of hydroxyproline in biological tissues. Clin Biochem 29: 225–229.
Reiter RJ, Tan DX, Mayo JC, Sainz RM, Leon J, Czarnocki Z (2003). Melatonin as an antioxidant: biochemical mechanisms and pathophysiological implications in humans. Acta Biochim Pol 50: 1129–1146.
Sainz RM, Mayo JC, Rodriguez C, Tan DX, Lopez-Burillo S, Reiter RJ (2003). Melatonin and cell death: Differential actions on apoptosis in normal and cancer cells. Cell Mol Life Sci 60: 1407–1426.
Schäfer M, Werner S (2008). Oxidative stress in normal and impaired wound repair. Pharmacol Res 58: 165–171.
Sener A, Ozsavci D, Bingol-Ozakpinar O, Cevik O, Yanikkaya- Demirel G, Yardimci T (2009). Oxidized-LDL and Fe3+/ascorbic acid-induced oxidative modifications and phosphatidylserine exposure in human platelets are reduced by melatonin. Folia Biol (Praha) 55: 45–52.
Sener G, Sert G, Sehirli AO, Arbak SN, Ayanoglu-Dulger G (2006). Melatonin protects against pressure ulcer-induced oxidative injury of the skin and remote organs in rats. J Pineal Res 40: 280–287.
Sharifi R, Rastegar RH, Kamalinejad M, Dehpour AR, Tavangar SM, Maliheh P, Natanzi MM, Ghannadian N, Akbari M, Pasalar P (2012). Effect of topical application of silymarin (Silybum marianum) on excision wound healing in albino rats. Acta Medica Iranica 50: 583-588.
Slominski A, Fischer TW, Zmijewski MA, Wortsman J, Semak I, Zbytek B, Slominski RM, Tobin DJ (2005). On the role of melatonin in skin physiology and pathology. Endocrine 27: 137–148.
Stratos I, Richte N, Rotter R, Li Z, Zechner D, Mittlmeier T, Vollmar B (2012). Melatonin restores muscle regeneration and enhances muscle function after crush injury in rats. J Pineal Res 52: 62–70.
Tomas-Zapico C, Coto-Montes A (2005). A proposed mechanism to explain the stimulatory effect of melatonin on antioxidative enzymes. J Pineal Res 39: 99–104.
Viswanathan AN, Schernhammer ES (2009). Circulating melatonin and the risk of breast and endometrial cancer in women. Cancer Lett 281: 1–7.
Vollmar B, El-Gibaly AM, Scheuer C, Strik MW, Bruch HP, Menger MD (2002). Acceleration of cutaneous wound healing by transient p53 inhibition. Lab Invest 82: 1063–1071.
Wu YS, Chen SN (2014). Apoptotic cell: linkage of inflammation and wound healing. Front Pharmacol 21: 1–6.
Yoo YM, Yim SV, Kim SS, Jang HY, Lea HZ, Hwang GC, Kim JW, Kim SA, Lee HJ, Kim CJ et al. (2002). Melatonin suppresses NO-induced apoptosis via induction of Bcl-2 expression in PGT-beta immortalized pineal cells. J Pineal Res 33: 146–150.