Influência do alcoolismo crônico e da deficiência estrogênica sobre a expressão imunoistoquímica de proteínas reguladoras do processo de reabsorção óssea no periodonto de ratas
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Data
2016-08-10
Autores
Marchini, Adriana Mathias Pereira da Silva [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Estudos prévios sugerem que tanto o consumo de álcool quanto a deficiência estrogênica poderiam afetar o osso alveolar e aumentar a suscetibilidade individual a uma mais rápida progressão da doença periodontal. No entanto, a literatura ainda não chegou a um consenso sobre esse assunto, que precisa ser mais estudado. O objetivo deste estudo foi investigar possíveis modificações na expressão imunoistoquímica de proteínas envolvidas na regulação do processo de reabsorção óssea, no periodonto de ratas submetidas ao alcoolismo crônico e/ou à deficiência estrogênica. Ratas Wistar, com três meses de idade, foram divididas em dois grupos: ovariectomizadas (ovz) ou não ovariectomizadas (sham). Trinta dias após os procedimentos cirúrgicos, os animais foram subdivididos e receberam os seguintes tratamentos: dieta alcoólica (alc), dieta isocalórica (iso) ou dieta livre (dl). Os grupos álcool receberam solução alcoólica (20%), por dois meses. A análise do tratamento dietético mostrou que o grupo ovz/dl foi o que mais ganhou peso e o que mais ingeriu ração, sendo as diferenças significativas se comparadas a todos os outros grupos (p<0,01). Quando considerados ambos os grupos álcool, 53,49% das calorias diárias da dieta (em média) foram provenientes do álcool. Os resultados também mostraram que o grupo sham/alc consumiu mais álcool se comparado ao ovz/alc, sendo essa diferença estatisticamente significativa (p<0,001). A crista óssea alveolar e tecidos adjacentes (na região entre o primeiro e segundo molar inferior) foram avaliados por imunoistoquímica, utilizando-se anticorpos para detecção de osteoprotegerina (OPG), proteína que inibe a reabsorção óssea, e ligante do receptor ativador do fator nuclear Kappa β (RANKL), proteína que estimula a reabsorção óssea. Os grupos ovz/alc, ovz/iso e ovz/dl apresentaram menor imunomarcação para a OPG, se comparados ao sham/alc, sham/iso e sham/dl, respectivamente, sendo essas diferenças estatisticamente significativas (p<0,05, p<0,05 e p<0,01). O grupo ovz/iso apresentou maior imunomarcação para o RANKL se comparado ao grupo sham/iso, sendo essa diferença estatisticamente significativa (p<0,01). O grupo sham/alc apresentou menor imunomarcação para a OPG e maior imunomarcação para o RANKL quando comparado aos outros grupos sham, sendo essas diferenças estatisticamente significativas (p<0,01). No grupo ovz/alc foi observada a menor imunomarcação para a OPG e a maior imunomarcação para o RANKL, sendo as diferenças estatisticamente significativas (p<0,05), quando comparado com a maioria dos outros grupos (exceção do ovz/iso e sham/alc, para o RANKL). Pode-se concluir que tanto a deficiência estrogênica quanto a dieta alcoólica foram relacionados à diminuição da expressão de OPG e aumento da expressão de RANKL, o que seria compatível com uma maior predisposição à reabsorção óssea. Esses resultados foram ainda mais evidentes no grupo no qual a deficiência estrogênica e o consumo de álcool foram associados.
Previous studies suggest that both chronic alcoholism and estrogen deficiency may affect alveolar bone and increase individual susceptibility to faster periodontal disease progression. However, the literature has not yet reached a consensus on this issue, which needs to be further studied. The aim of this study was to investigate possible changes in immunohistochemical expression of proteins involved in the regulation of bone resorption process in the periodontium of female rats subjected to chronic alcoholism and/or estrogen deficiency. Wistar female rats, at three months of age, were divided into two groups: ovariectomized (ovx) or not ovariectomized (sham). Thirty days after the surgical procedures, animals were split again and received the following treatments: alcoholic diet (alc), isocaloric diet (iso) or ad libitum diet (ad). The alcohol groups received alcoholic solution (20%), for two months. The dietary treatment analysis showed ovx/ad group gained more weight and ingested more solid food and these differences were statistically significant as compared with all other groups (p<0.01). When considering both alcohol groups, 53.49% of the daily calories diet (in average) were coming from alcohol. The results also showed the sham/alc group ingested more alcohol as compared to ovx/alc group and this difference was statistically significant (p<0.001).The alveolar bone crest and adjacent tissues (in the region between first and second lower molars) were evaluated by immunohistochemistry, using antibodies for detection of osteoprotegerin (OPG), a protein that inhibits bone resorption, and receptor activator of nuclear factor-kappa β ligand (RANKL), a protein that stimulates bone resorption. The groups ovx/alc, ovx/iso and ovx/ad showed lower immunostaining for OPG, compared to sham/alc sham/iso and sham/ad, respectively, and these differences were statistically significant (p<0.05, p<0.05, and p<0.01). The group ovx/iso showed higher immunostaining for the RANKL compared to sham/iso group, and this difference was statistically significant (p<0.01). Sham/alc group showed lower immunostaining for OPG and higher immunostaining for the RANKL when compared to other sham groups, and these differences were statistically significant (p<0.01). In ovx/alc group the lowest immunostaining for OPG and the highest immunostaining for the RANKL were observed, and these differences were statistically significant (p<0.05) as compared to the most others groups (except ovx/iso and sham/alc for RANKL). It can be concluded that estrogen deficiency and alcohol diet were related to decreased expression of OPG and increased expression of RANKL, which would be consistent with a greater predisposition to bone resorption. These results were even more evident in the group in which estrogen deficiency and alcohol consumption were associated.
Previous studies suggest that both chronic alcoholism and estrogen deficiency may affect alveolar bone and increase individual susceptibility to faster periodontal disease progression. However, the literature has not yet reached a consensus on this issue, which needs to be further studied. The aim of this study was to investigate possible changes in immunohistochemical expression of proteins involved in the regulation of bone resorption process in the periodontium of female rats subjected to chronic alcoholism and/or estrogen deficiency. Wistar female rats, at three months of age, were divided into two groups: ovariectomized (ovx) or not ovariectomized (sham). Thirty days after the surgical procedures, animals were split again and received the following treatments: alcoholic diet (alc), isocaloric diet (iso) or ad libitum diet (ad). The alcohol groups received alcoholic solution (20%), for two months. The dietary treatment analysis showed ovx/ad group gained more weight and ingested more solid food and these differences were statistically significant as compared with all other groups (p<0.01). When considering both alcohol groups, 53.49% of the daily calories diet (in average) were coming from alcohol. The results also showed the sham/alc group ingested more alcohol as compared to ovx/alc group and this difference was statistically significant (p<0.001).The alveolar bone crest and adjacent tissues (in the region between first and second lower molars) were evaluated by immunohistochemistry, using antibodies for detection of osteoprotegerin (OPG), a protein that inhibits bone resorption, and receptor activator of nuclear factor-kappa β ligand (RANKL), a protein that stimulates bone resorption. The groups ovx/alc, ovx/iso and ovx/ad showed lower immunostaining for OPG, compared to sham/alc sham/iso and sham/ad, respectively, and these differences were statistically significant (p<0.05, p<0.05, and p<0.01). The group ovx/iso showed higher immunostaining for the RANKL compared to sham/iso group, and this difference was statistically significant (p<0.01). Sham/alc group showed lower immunostaining for OPG and higher immunostaining for the RANKL when compared to other sham groups, and these differences were statistically significant (p<0.01). In ovx/alc group the lowest immunostaining for OPG and the highest immunostaining for the RANKL were observed, and these differences were statistically significant (p<0.05) as compared to the most others groups (except ovx/iso and sham/alc for RANKL). It can be concluded that estrogen deficiency and alcohol diet were related to decreased expression of OPG and increased expression of RANKL, which would be consistent with a greater predisposition to bone resorption. These results were even more evident in the group in which estrogen deficiency and alcohol consumption were associated.
Descrição
Palavras-chave
Alcoolismo, Ovariectomia, Osteoporose, Periodonto, Reabsorção óssea, Alcoholism, Ovariectomy, Osteoporosis, Periodontium, Bone Resorption