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Evaluation of antioxidant and anti-inflammatory effect of the lupeol-based cream isolated from Bowdichia virgilioides Kunth (Sucupira black) on the cutaneous wound healing

Grant number: 14/23441-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): March 01, 2015
Effective date (End): February 29, 2016
Field of knowledge:Biological Sciences - Pharmacology
Principal Investigator:Claudia Helena Pellizzon
Grantee:Danilo Augusto Bérgamo
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The difficulty in wound healing is a major complication of diabetes mellitus (DM), which makes the healing process slower and more costly treatment. This process is divided into three overlapping phases: the inflammatory phase, the main events are hemostas is and inflammation, proliferative phase in which there is the formation of granulation tissue and phase extracellular matrix remodeling. It is known that the high level of oxidative stress and the persistence of the inflammatory process contributing to delayed healing of skin wounds. The Lupeol is a triterpene found in plant Bowdichia virgilioides Kunth (Sucupira black) and has several proven biological activities. Therefore, this project aims to investigate the effect and mechanisms involved in the healing action of the cream-based lupeol isolated from the stem bark of B. virgiliodes Kunth in skin wounds in rats. To this end, male Wistar rats (n = 8) will be randomly divided into the following experimental groups: sham group (no treatment and induction of the wound) cream base (negative control), collagenase (positive control), based cream lupeol 0, 1%, 0.2% and 0.4% (test substance). The animals will be anesthetized, and subsequently the manufacture of wounds in the dorsal region with the aid of a punch 2 cm in diameter. The lesions will be photographed once a day for 3, 7 or 14 days (according to the phases described in the literature) and measured with the aid of specific software ImageJ. After euthanasia, the region of the wound will be designed to histological analysis (count of inflammatory cells, inflammatory score, morphology and deposition of collagen fibers), morphometric (height of the epithelium regeneration, thickness of epidermis and dermis). The antioxidant activity is determined by measuring levels of reduced glutathione (GSH), glutathione reductase (GR) and peroxidase (GPx), superoxide dismutase (SOD) and myeloperoxidase (MPO) and the anti-inflammatory activity by the method of gel zymography to determine the enzymatic activity of matrix metalloproteinases MMP-2 and MMP-9 This research is linked to PhD project "Evaluation of the cellular and molecular mechanisms of lupeolbased cream on the process of cutaneous wound healing in experimental models ofdiabetic and nondiabetic rats" and the research assistance project approved by FAPESP "Healing process in skin under treatment with monoterpene (Menthol) and triterpene (Lupeol) in experimental models" process 2013/23340-1, which contributed significantly to the implementation of this research in the laboratory as well as in the training of human resources.