Evaluation of the cellular and molecular mechanisms of lupeol-based cream on the process of cutaneous wound healing in experimental models of diabetic and nondiabetic rats

• Grant number: 14/23247-4
• Support Opportunities: Scholarships in Brazil - Doctorate
• Effective date (Start): November 01, 2016
• Effective date (End): 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)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BESERRA, FERNANDO PEREIRA; VIEIRA, ANA JULIA; SERGIO GUSHIKEN, LUCAS FERNANDO; DE SOUZA, EDUARDO OLIVEIRA; HUSSNI, MARIA FERNANDA; HUSSNI, CARLOS ALBERTO; NOBREGA, RAFAEL HENRIQUE; MONTEIRO MARTINEZ, EMANUEL RICARDO; JACKSON, CHRISTOPHER JOHN; DE AZEVEDO MAIA, GABRIELA LEMOS; et al. Lupeol, a Dietary Triterpene, Enhances Wound Healing in Streptozotocin-Induced Hyperglycemic Rats with Modulatory Effects on Inflammation, Oxidative Stress, and Angiogenesis. OXIDATIVE MEDICINE AND CELLULAR LONGEVITY, 2019. (13/23340-1, 14/23247-4, 17/17600-1)

BESERRA, FERNANDO PEREIRA; VIEIRA, ANA JULIA; SERGIO GUSHIKEN, LUCAS FERNANDO; DE SOUZA, EDUARDO OLIVEIRA; HUSSNI, MARIA FERNANDA; HUSSNI, CARLOS ALBERTO; NOBREGA, RAFAEL HENRIQUE; MONTEIRO MARTINEZ, EMANUEL RICARDO; JACKSON, CHRISTOPHER JOHN; DE AZEVEDO MAIA, GABRIELA LEMOS; et al. Lupeol, a Dietary Triterpene, Enhances Wound Healing in Streptozotocin-Induced Hyperglycemic Rats with Modulatory Effects on Inflammation, Oxidative Stress, and Angiogenesis. OXIDATIVE MEDICINE AND CELLULAR LONGEVITY, v. 2019, p. 20-pg., 2019-01-01. (14/23247-4, 17/17600-1, 13/23340-1)

BESERRA, FERNANDO PEREIRA; SERGIO GUSHIKEN, LUCAS FERNANDO; VIEIRA, ANA JULIA; BERGAMO, DANILO AUGUSTO; BERGAMO, PATRICIA LUISA; DE SOUZA, MARIANA OLIVEIRA; HUSSNI, CARLOS ALBERTO; TAKAHIRA, REGINA KIOMI; NOBREGA, RAFAEL HENRIQUE; MONTEIRO MARTINEZ, EMANUEL RICARDO; et al. From Inflammation to Cutaneous Repair: Topical Application of Lupeol Improves Skin Wound Healing in Rats by Modulating the Cytokine Levels, NF-kappa B, Ki-67, Growth Factor Expression, and Distribution of Collagen Fibers. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 21, n. 14, JUL 2020. (13/23340-1, 14/23247-4, 17/17600-1)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

BESERRA, Fernando Pereira. Mechanisms involved in the cutaneous wound healing effect of lupeol isolated from the stem bark of Bowdichia virgilioides Kunth. (Fabaceae) in experimental models in vivo and in vitro. 2019. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Instituto de Biociências. Botucatu Botucatu.