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Doctoral Thesis
DOI
Document
Author
Full name
Mayara Sgarbi Semeghini
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2019
Supervisor
Committee
Prado, Karina Fittipaldi Bombonato (President)
Martins, Manoela Domingues
Okamoto, Roberta
Siessere, Selma
Title in Portuguese
Efeito do carotenoide licopeno no metabolismo ósseo e na atividade funcional de células da linhagem osteogênica de ratas ovariectomizadas
Keywords in Portuguese
Carotenoide
Licopeno
Metabolismo ósseo
Osteoporose
Remodelação óssea
Abstract in Portuguese
A osteoporose é uma doença silenciosa caracterizada pela diminuição da massa óssea levando à morbidade e alto risco de fratura. A sua incidência é maior em mulheres após a menopausa devido à mudanças hormonais, assim como propensão genética e hábitos de vida como sedentarismo, alcoolismo, tabagismo e alimentação inadequada. Além de alimentos ricos em cálcio que favorecem a manutenção da massa e densidade óssea, existem outros nutrientes que podem influenciar na prevenção de doenças associadas com o metabolismo ósseo, como os carotenoides. Entre os carotenoides, o licopeno tem sido associado a estes efeitos principalmente devido à suas propriedades antioxidantes. Assim, o objetivo desta investigação foi realizar estudos in vivo e in vitro sobre o efeito do licopeno no metabolismo ósseo de ratas osteoporóticas. Ratas Wistar foram ovariectomizadas e pareadas com animais sham. As avaliações in vitro foram realizadas após 60 dias da cirurgia, quando as células foram cultivadas em meio osteogênico e divididas em grupos controle (sham), ovariectomizado (Ovx) e ovariectomizado + 1 µmol/L de licopeno (Ovx/Lic). Após os períodos experimentais, foram avaliados proliferação celular, detecção in situ de fosfatase alcalina, mineralização e expressão quantitativa de genes associados ao metabolismo ósseo. Além disso, os estudos in vivo foram realizados para avaliar a remodelação óssea por meio de parâmetros tomográficos, histomorfométricos e estereológicos após a ingestão diária de 10 mg/kg de licopeno por 60 dias após a ovariectomia. Todos os dados quantitativos foram analisados por meio de teste estatístico ANOVA com significância para p ≤ 0.05. Os resultados in vitro mostraram que a presença do licopeno favoreceu a proliferação celular e a detecção in situ de ALP em estágio tardios das culturas, assim como promoveu um aumento significativo na deposição de nódulos mineralizados. O licopeno também aumentou a expressão dos genes Alp, Runx2, Bglap e Bmp4, além de diminuir a razão Rankl/Opg quando comparado ao grupo Ovx. Os experimentos in vivo mostraram que a administração oral do licopeno promoveu uma significativa manutenção do volume tecidual e volume ósseo na epífise femoral, assim como do trabeculado ósseo e quantidade de osteócitos. Os resultados sugerem que o licopeno pode beneficiar o metabolismo do tecido ósseo associado à osteoporose
Title in English
Effect of carotenoid lycopene on bone metabolism and functional activity of cells from the osteogenic lineage of ovariectomized rats
Keywords in English
Bone metabolism
Bone remodeling
Carotenoids
Lycopene
Osteoporosis
Abstract in English
Osteoporosis is a silence disease characterized by a decrease in bone mass leading to morbidity and high risk of fractures. Its incidence is higher in women after menopause because of hormonal changes, as well as genetic propension and life styles such as sedentarism, alcoholism, tabagism and poor food intake. Besides calcium rich foods that favour the maintenance of bone mass and its density, there are several other nutrients that may influence in the prevention of diseases associated to bone metabolism, such as carotenoids. Among carotenoids, lycopene has been associated to prevention of bone diseases because of its antioxidant properties. The purpose of this investigation was to perform in vitro and in vivo studies about the effect of lycopene in bone metabolism of ovariectomized rats. Wistar rats were ovariectomized and paired with sham animals. In vitro evaluations were performed after 60 days of surgery, when cells were cultured in osteogenic medium and divided into control (sham), ovariectomized (Ovx) and ovariectomized + 1 µmol/L lycopene (Ovx/Lic) groups. After the experimental periods, there were evaluated cell proliferation, alkaline phosphatase in situ detection, mineralization and quantitative expression of genes associated to bone metabolism. Besides, in vivo studies were carried out to evaluate femur remodeling by tomographic, histomorphometric and estereological evaluation after daily intake of 10 mg/kg of lycopene for 60 days after ovariectomy. All quantitative data were analyzed by means of one-way ANOVA statistical test with significance of p ≤ 0.05. The in vitro results showed that the presence of lycopene favored cell proliferation and in situ detection of ALP in late culture stages, as well as promoted a significative increase in the deposition of mineralized nodules. Lycopene significatively increased the expression of genes Alp, Runx2, Bglap and Bmp4, besides decreasing the Rankl/Opg ratio when compared to Ovx group. In vivo experiments showed that oral administration of lycopene promoted a significant maintenance of tissue and bone volume in femoral epiphysis, as well as of bone trabeculae and number of osteocytes. Results suggest that lycopene can benefit bone metabolism with the onset of osteoporosis
 
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Publishing Date
2019-10-17
 
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