Hemin attenuated oxidative stress and inflammation to improve wound healing in diabetic rats
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Oxidative stress and persistent inflammation play crucial role in the progression of diabetic wound complications. Hemeoxgenase-1 (HO-1) by degrading hemin has been shown to display anti-oxidant and anti-inflammatory effects. Further, hemin is a potent HO-1 inducer. Thus, the current study was aimed to evaluate the effect of topical application of hemin on diabetic wound in rats. Four hundred square millimeter open excision wound were created 2 weeks after induction of diabetes with single intraperitoneal injection of streptozotocin (60 mg/kg), and the diabetic rats were divided into three groups namely diabetic control, hemin, and tin protoporphyrin (SnPPIX). Ointment base, hemin (0.5% in ointment base), and SnPPIX (0.5% in ointment base) were applied topically to wounded area in diabetic control, hemin, and SnPPIX group rats, respectively, twice daily for 19 days. Hemin significantly increased the wound contraction in comparison to control and SnPPIX-treated rats. Time-dependent analysis revealed significant increase in anti-oxidants with concomitant decrease in oxidants in hemin-treated rats as compared to diabetic control rats. Further, mRNA expression decreased for inflammatory cytokine and increased for anti-inflammatory cytokine in hemin group as compared to diabetic control rats. Expression of HO-1 also increased in hemin group as compared to diabetic control rats. However, SnPPIX group results were in disagreement with results of hemin which is clearly reflected in histopathology. Results indicate the ability of hemin to accelerate wound healing in diabetic rats by combating inflammation and oxidative stress probably via HO-1.
KeywordsDiabetes Wound Hemin Tin protoporphyrin Oxidative stress Inflammation
DK, MR, VK, and DK conceived and designed the research program. DK, MCL, VS, RP, and SK conducted the experiments. RP, VK, and PG conducted the mRNA studies. DK, GRJ, and SKT analyzed the data. DK and SKT interpreted the results. DK, GRJ, and DK wrote the manuscript. All authors read and approved the manuscript.
The Director the Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India, provided the necessary funds and facilities for conducting the present study to the first author.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All the experimental procedures conducted on animals (Wistar rats) were in compliance with at least the Declaration of the National Institutes of Health Guide for Care and Use of Laboratory Animals. The experimental procedures and protocols were duly approved by the Institutional Animal Ethics Committee of Indian Veterinary Research Institute, Izatnagar, India.
This article does not contain any studies with human participants performed by any of the authors.
- Berlanga-Acosta J, Valdéz-Pérez C, Savigne-Gutiérrez W, Mendoza Mari Y, Franco Pérez N, Vargas Machiran E, Poll Marrón N, Alvarez Duarte H, Echeverría Requeijo H, Pérez Aguilar RM (2010) Cellular and molecular insights into the wound healing mechanism in diabetes. Biotecnología Aplicada 27:255–261Google Scholar
- Fadini GP, Albiero M, Menegazzo L, Boscaro E, Pagnin E, Iori E, Cosma C, Lapolla A, Pengo V, Stendardo M, Agostini C, Pelicci PG, Giorgio M, Avogaro A (2010) The redox enzyme p66Shc contributes to diabetes and ischemia-induced delay in cutaneous wound healing. Diabetes 59:2306–2314. https://doi.org/10.2337/db09-1727 CrossRefGoogle Scholar
- Greenhalgh DG, Sprugel KH, Murray MJ, Ross R (1990) PDGF and FGF stimulate healing in the genetically diabetic mouse. Am J Pathol 136:1235–1246Google Scholar
- Grochot-Przeczek A, Lach R, Mis J, Skrzypek K, Gozdecka M, Sroczynska P, Dubiel M, Rutkowski A, Kozakowska M, Zagorska A, Walczynski J, Was H, Kotlinowski J, Drukala J, Kurowski K, Kieda C, Herault Y, Dulak J, Jozkowicz A (2009) Heme oxygenase-1 accelerates cutaneous wound healing in mice. PLoS One 4:e5803. https://doi.org/10.1371/journal.pone.0005803 CrossRefGoogle Scholar
- Hyvelin JM, Maurel B, Uzbekov R, Motterlini R, Lermusiaux P (2010) Hemin prevents in-stent stenosis in rat and rabbit models by inducing heme-oxygenase-1. J Vasc Surg 51:417–428. https://doi.org/10.1016/j.jvs.2009.09.004
- Juckett MB, Balla J, Balla G, Jessurun J, Jacob HS, Vercellotti GM (1995) Ferritin protects endothelial cells from oxidized low density lipoprotein in vitro. Am J Pathol 147:782–789Google Scholar
- Khanna S, Biswas S, Shang Y, Collard E, Azad A, Kauh C, Bhasker V, Gordillo GM, Sen CK, Roy S (2010) Macrophage dysfunction impairs resolution of inflammation in the wounds of diabetic mice. PLoS One 5:e9539. https://doi.org/10.1371/journal.pone.0009539
- Madesh M, Balasubramanian KA (1998) Microtiter plate assay for superoxide dismutase using MTT reduction by superoxide. Indian J Biochem Biophys 35(3):184–188Google Scholar
- Morse D, Choi AM (2005) Heme oxygenase-1: from bench to bedside. Am J Respir Crit Care Med 172:660–670. https://doi.org/10.1164/rccm.200404-465SO
- Ndisang JF, Lane N, Jadhav A (2009) The heme oxygenase system abates hyperglycemia in Zucker diabetic fatty rats by potentiating insulin-sensitizing pathways. Endocrinology 150:2098–2108. https://doi.org/10.1210/en.2008-0239
- Ram M, Singh V, Kumar D, Kumawat S, Gopalakrishnan A, Lingaraju MC, Gupta P, Tandan SK, Kumar D (2014) Antioxidant potential of bilirubin-accelerated wound healing in streptozotocin-induced diabetic rats. Naunyn Schmiedeberg's Arch Pharmacol 387:955–961. https://doi.org/10.1007/s00210-014-1011-3 CrossRefGoogle Scholar
- Schonfelder U, Abel M, Wiegand C, Klemm D, Elsner P, Hipler UC (2005) Influence of selected wound dressings on PMN elastase in chronic wound fluid and their antioxidative potential in vitro. Biomaterials 26:6664–6673. https://doi.org/10.1016/j.biomaterials.2005.04.030 CrossRefGoogle Scholar
- Shukla A, Rasik AM, Patnaik GK (1997) Depletion of reduced glutathione, ascorbic acid, vitamin E and antioxidant defence enzymes in a healing cutaneous wound. Free Radic Res 26:93–101Google Scholar
- Taye A, Ibrahim BM (2013) Activation of renal haeme oxygenase-1 alleviates gentamicin-induced acute nephrotoxicity in rats. J Pharm Pharmacol 65:995–1004. https://doi.org/10.1111/jphp.12067
- Wetzler C, Kampfer H, Stallmeyer B, Pfeilschifter J, Frank S (2000) Large and sustained induction of chemokines during impaired wound healing in the genetically diabetic mouse: prolonged persistence of neutrophils and macrophages during the late phase of repair. J Invest Dermatol 115:245–253. https://doi.org/10.1046/j.1523-1747.2000.00029.x CrossRefGoogle Scholar