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Master's Dissertation
DOI
https://doi.org/10.11606/D.58.2019.tde-29082018-100641
Document
Author
Full name
Maria Cecilia Gorita dos Santos
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2018
Supervisor
Committee
Issa, João Paulo Mardegan (President)
Bezerra, Thiago Augusto Rochetti
Fazan, Valeria Paula Sassoli
Segato, Raquel Assed Bezerra
Title in Portuguese
Avaliação óssea, histológica e imuno-histoquímica em modelo animal ovariectomizado e aterosclerótico
Keywords in Portuguese
Aterosclerose
MMP-9
Osteoporose
Abstract in Portuguese
A osteoporose é uma doença osteometabólica que atinge milhares de pessoas; e as doenças cardiovasculares são as principais causas de mortes no mundo. Estudos demonstram que pacientes com baixa densidade mineral óssea apresentam maior risco de desenvolverem doenças cardiovasculares. No entanto, os mecanismos patológicos, fisiológicos e moleculares que correlacionam a osteoporose e a aterosclerose ainda são desconhecidos. O objetivo deste estudo in vivo foi realizar avaliação histológica e imuno-histoquímica do osso fêmur de camundongos ovariectomizados e ateroscleróticos após tratamento com três drogas: doxiciclina, atorvastatina e vitamina D. Foram utilizados 36 camundongos fêmeas, como controle (linhagem C57BL/6), divididos aleatoriamente em seis grupos: Grupo I: Água; Grupo II: Etanol; Grupo III: Azeite; Grupo IV: Doxiciclina; Grupo V: Atorvastatina e Grupo VI: Vitamina D; e 36 camundongos fêmeas knockout para Apolipoproteína E (ApoE-/-) divididos aleatoriamente em seis grupos: Grupo VII: Água OVX; Grupo VIII: Etanol OVX; Grupo IX: Azeite OVX; Grupo X: Doxiciclina OVX; Grupo XI: Atorvastatina OVX e Grupo XII: Vitamina D OVX. Após indução dos quadros de osteoporose e aterosclerose foi realizado o tratamento conforme cada grupo. Decorrido o tempo experimental, os animais foram eutanasiados e o material coletado submetido ao processamento histotécnico para análise histológica, em cortes corados com Tricrômio de Masson e avaliação da expressão por imunohistoquímica de TRAP, IL-1β, TNF-α, RANKL e MMP-9. Os resultados obtidos foram submetidos à análise estatística por meio dos testes One-Way ANOVA, seguido pelo pósteste de Bonferroni (histologia e colesterol); Kruskal Wallis e Student-Newman-Keuls (imuno-histoquímica). O nível de significância adotado foi de 5%. Os resultados obtidos evidenciaram diferença significativa entre o número de células imunorreativas para MMP-9, TRAP, IL-1β, TNF-α e RANKL nos grupos VII e X; VIII e XI; demonstrando a ação da Doxiciclina e Atorvastatina sob tais moléculas. Os resultados histológicos evidenciaram a eficácia da vitamina D no tratamento da osteoporose, uma vez que promoveu aumento da área óssea
Title in English
Bone, histological and immunohistochemical evaluation in an ovariectomized and atherosclerotic animal model
Keywords in English
Atherosclerosis
MMP-9
Osteoporosis
Abstract in English
Osteoporosis is an osteometabolic disease that affects thousands of people; and cardiovascular diseases are the leading causes of death in the world. Studies have shown that patients with low bone density are at a higher risk of developing cardiovascular disease. However, the pathological, physiological and molecular mechanisms that are correlated to osteoporosis and atherosclerosis are still unknown. The present work in vivo was to perform a histological and immunohistochemical evaluation of the femoral bone of ovariectomized and atherosclerotic mice after treatment with three drugs: doxycycline, atorvastatin and vitamin D. Thirty-six female mice were used as controls (C57BL/6 strain) randomly divided into six groups: Group I: Water; Group II: Ethanol; Group III: Olive oil; Group IV: Doxycycline; Group V: Atorvastatin and Group VI: Vitamin D; and the 36 knockout mice for Apolipoprotein E (ApoE-/-) randomly divided into six groups: Group VII: Water OVX; Group VIII: Ethanol OVX; Group IX: OVX olive oil; Group X: Doxycycline OVX; Group XI: Atorvastatin OVX and Group XII: Vitamin D OVX. After induction of osteoporosis and atherosclerosis, treatment was performed according to each group. After the experimental time, the animals were euthanized and the collected material submitted to histotechnical processing for histological analysis in stained sections with Masson's Trichrome and evaluation of the expression by immunohistochemistry of TRAP, IL-1β, TNF-α, RANKL and MMP-9. The results were submitted to statistical analysis using One-Way ANOVA, followed by Bonferroni post-test (histology and cholesterol); Kruskal Wallis and Student-Newman-Keuls (immunohistochemistry). The level of significance was 5%. The results obtained showed a significant difference between the number of immunoreactive cells for MMP-9, TRAP, IL-1β, TNF-α and RANKL in groups VII and X; VIII and XI; demonstrating the action of Doxycycline and Atorvastatin on such molecules. The histological results evidenced the efficacy of vitamin D in the treatment of osteoporosis, since it promoted increased bone area
 
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Publishing Date
2019-07-19
 
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