| Grant number: | 14/23247-4 |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| Start date: | November 01, 2016 |
| End date: | March 31, 2019 |
| Field of knowledge: | Health Sciences - Pharmacy - Pharmacognosy |
| Principal Investigator: | Claudia Helena Pellizzon |
| Grantee: | Fernando Pereira Beserra |
| Host Institution: | Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil |
| Associated scholarship(s): | 18/06042-0 - Interaction between Vitamin D analogs and anticancer drug (cediranib) in human malignant melanoma cell lines, BE.EP.DR |
Abstract Wound healing difficulty is one of the major complications of diabetes mellitus (DM), which makes the healing process slower and the treatment more expensive. This process is divided into three overlapping phases: inflammatory phase (Which the main events are hemostasis and inflammation), proliferative phase (there is the formation of granulation tissue) and the phase of extracellular matrix remodeling. It is known that the high level of oxidative stress and the persistence of inflammation contribute to delayed healing of skin wounds in diabetic patients. The Lupeol is a triterpene found in many vegetables such as figs, mango, carrot, soybean, melon, grapes, propolis and has several proven biological activities. Thus, this project aims to investigate the effect and mechanisms involved in the healing action of lupeol-based cream on skin wounds in diabetic rats (DR) and non-diabetic rats (RND). Male Wistar rats (n=8) will be randomly divided into the following experimental groups: base cream (control RND), collagenase (positive control RND) and lupeol-based cream 0,1 %, 0,2 % and 0,4 in RND, cream base (control RD), insulin-based cream (positive control RD) and lupeol-based cream (best effective dose) in RD. The experimental induction of diabetes will be performed by a single intraperitoneal application of streptozotocin at a dose of 55 mg/kg. After the confirmation of the diabetic state of the animals, the rats will be anesthetized and the wounds induced in the dorsal region with the aid of a punch than 2 cm in diameter. The lesions will be treated and measured daily for 3, 7 or 14 days (according to the phases described in the literature). After the death, the region of the wound will be devoted to histological analysis (inflammatory cells count, inflammatory score, morphology of collagen fibers), histomorphometrical (regenerative epithelium height measurement) and immunohistochemistry (epidermal and vascular endothelial growth factors, cell proliferation and inflammatory response), determination of collagen and besides molecular analyzes (qPCR and RT-PCR to collagen I and III and epidermal, ²1-transforming growth factor). Antioxidant activity (reduced glutathione, glutathione reductase and peroxidase, superoxide dismutase, myeloperoxidase and nitrite), anti-inflammatory activity by the method of gel zymography to determine the enzymatic activity of matrix metalloproteinases MMP-2 and MMP-9 and determination of cytokine levels in serum by ELISA (TNF-±, IL-1² and IL-6 and IL-10) will be performed. This research is linked to the funding project "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. (AU) | |
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