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Effects of smoking and alcohol moderate intake on the genes expression of ubiquitin-proteasome proteolytic pathway

Grant number: 14/02267-7
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2014
Effective date (End): December 31, 2014
Field of knowledge:Health Sciences - Physiotherapy and Occupational Therapy
Principal researcher:Patricia Monteiro Seraphim
Grantee:Tamara dos Santos Gouveia
Home Institution: Faculdade de Ciências e Tecnologia (FCT). Universidade Estadual Paulista (UNESP). Campus de Presidente Prudente. Presidente Prudente , SP, Brazil

Abstract

The behavior of society has beend modified due to increase of industrialization, with greater exposure to risk factors that impairs health of people, such as smoking and alcohol intake. Both cause major diseases, such as COPD (chronic obstructive pulmonary disease) and alcoholic myopathy, which interferes in the musculoskeletal system, causing muscle atrophy and initiates the process of proteolysis. This process is regulated by the signaling pathway ubiquitin- proteasome through the expression of proteins like ubiquitin (coded by Ubb gene) and E214kDa (coded by E214kda) and MuRF -1 (coded by Trim63). But, it has been confirmed in numerous studies, that physical exercise can reduce the process of muscle atrophy. Thus the aim of the study is to evaluate the effect of a program of smoking, alcohol and exercise training in the EDL (Extensor digitorum longus) muscle of male Wistar rats on the analysis of the expression of Ubb, Ube2b and Trim63 genes. The animals will be kept in the bioterium of Department of Physical Therapy, Faculty of Science and Technology, UNESP - Presidente Prudente and they will be randomly divided into eight groups of 10 rats per group: C (control group), S (smoking group), A (alcohol group) , SA (smoking alcoholic group) , E (exercised group), SE (smoking exercised group), AE (alcoholic exercised group) , ASE (alcoholic smoking exercised group). The smoking groups will be submitted to a protocol of exposure to cigarette smoke with combustion of 4 cigarettes for 30 minutes twice a day, five days a week, for 60 days. The alcoholic groups will intake ethanol at 10% v/v concentration for 60 days. The training program will be performed in a treadmill with sessions of five times a week, for 60 days, 60 minutes each session, after adaptation. After 24 hours the last training session, the animals will be sacrificed using anesthetic Ketamine and Xylazine (40 mg / kg body weight) intraperitoneally. Then the EDL skeletal muscle will be removed. RT-PCR wil be performed for Ubb, Ube2b, and Trim63 genes expression. ANOVA test, parametric and post-test will be done if necessary for comparison of data. The difference among groups will be considered significant when P <0.05.

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