The impact of testosterone on aging-related remodeling of salivary glands

Detalhes bibliográficos
Autor(a) principal: Santos, João Miguel Valente
Data de Publicação: 2023
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10773/39618
Resumo: Salivary glands play an important role in the secretion of saliva into the oral cavity, influencing salivary composition and oral health. However, the molecular profile of salivary glands is poorly understood, especially in clinical scenarios associated with changes in sex hormone levels. Therefore, our study aimed to investigate how chronic exposure to supraphysiological testosterone levels affects salivary gland remodelling, with a particular focus on proteome adaptation. To achieve this goal, male Wistar rats were implanted subcutaneously at 12 weeks-of-age with devices that release testosterone. At 35 and 61 weeks-of-age, the animals were sacrificed, and their salivary glands were harvested for histological and molecular characterization. The data showed that the submandibular glands were more sensitive to testosterone compared with the parotid glands, as evidenced primarily by a significantly higher ratio of gland mass to body weight. Further analysis of the submandibular glands revealed histologic evidence of acinar cell hypertrophy. At the molecular level, no changes in tissue levels of androgen receptor (AR) and estrogen receptor alpha were detected despite significantly increased levels of circulating sex hormones. GeLC- MS /MS analysis of submandibular gland extracts identified 308 different proteins, 12 of which were modulated by testosterone. Of the common proteins identified from the CTRL and TEST groups, 14 were up-regulated by testosterone, but no relevant interactions were observed between them. In addition, zymographic analysis of the submandibular glands revealed a similar proteolytic profile between the CTRL and TEST groups, with no significant differences observed in optical density values between the two groups. However, our data highlight the contribution of kallikreins (Klk) to the proteolytic activity of these glands, as seen in the zymography gel profile. When looking at the individual proteins that were up-regulated in the TEST group, the stronger expression of Klk3 in the mandibular glands of TEST rats was striking (2-fold increase compared with the CTRL group). Moreover, this protease was also the only kallikrein that showed interaction with the AR, suggesting that Klk3 is a target of this receptor. These results shed light on the differential response of salivary glands to testosterone exposure and provide insight into the proteome-level changes associated with this exposure. Understanding the molecular adaptations in the salivary glands may contribute to a better understanding of oral health and the development of potential therapeutic interventions.
id RCAP_ef2590da22d7e48d93a10c6fa33d82bd
oai_identifier_str oai:ria.ua.pt:10773/39618
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling The impact of testosterone on aging-related remodeling of salivary glandsSalivary glandsSubmandibular gland’s proteomeTestosteroneAgingRatsGeLC-MS/MSSalivary glands play an important role in the secretion of saliva into the oral cavity, influencing salivary composition and oral health. However, the molecular profile of salivary glands is poorly understood, especially in clinical scenarios associated with changes in sex hormone levels. Therefore, our study aimed to investigate how chronic exposure to supraphysiological testosterone levels affects salivary gland remodelling, with a particular focus on proteome adaptation. To achieve this goal, male Wistar rats were implanted subcutaneously at 12 weeks-of-age with devices that release testosterone. At 35 and 61 weeks-of-age, the animals were sacrificed, and their salivary glands were harvested for histological and molecular characterization. The data showed that the submandibular glands were more sensitive to testosterone compared with the parotid glands, as evidenced primarily by a significantly higher ratio of gland mass to body weight. Further analysis of the submandibular glands revealed histologic evidence of acinar cell hypertrophy. At the molecular level, no changes in tissue levels of androgen receptor (AR) and estrogen receptor alpha were detected despite significantly increased levels of circulating sex hormones. GeLC- MS /MS analysis of submandibular gland extracts identified 308 different proteins, 12 of which were modulated by testosterone. Of the common proteins identified from the CTRL and TEST groups, 14 were up-regulated by testosterone, but no relevant interactions were observed between them. In addition, zymographic analysis of the submandibular glands revealed a similar proteolytic profile between the CTRL and TEST groups, with no significant differences observed in optical density values between the two groups. However, our data highlight the contribution of kallikreins (Klk) to the proteolytic activity of these glands, as seen in the zymography gel profile. When looking at the individual proteins that were up-regulated in the TEST group, the stronger expression of Klk3 in the mandibular glands of TEST rats was striking (2-fold increase compared with the CTRL group). Moreover, this protease was also the only kallikrein that showed interaction with the AR, suggesting that Klk3 is a target of this receptor. These results shed light on the differential response of salivary glands to testosterone exposure and provide insight into the proteome-level changes associated with this exposure. Understanding the molecular adaptations in the salivary glands may contribute to a better understanding of oral health and the development of potential therapeutic interventions.As glândulas salivares desempenham um papel importante na secreção de saliva para a cavidade oral, influenciando a saúde oral. No entanto, o perfil molecular das glândulas salivares está pouco estudado, principalmente associado a alterações nos níveis das hormonas sexuais. Assim, a presente dissertação teve como objetivo investigar de que forma a exposição crónica a níveis suprafisiológicos de testosterona afeta a remodelação das glândulas salivares, com particular foco na adaptação do proteoma. Para atingir este objetivo, foram implantados subcutaneamente em ratos Wistar machos, com 12 semanas de idade, dispositivos que libertam testosterona. Às 35 e 61 semanas de idade, os animais foram sacrificados e as suas glândulas salivares foram removidas para caracterização histológica e molecular. Os resultados mostraram que a glândula submandibular era mais sensível à testosterona em comparação com a parótida, como evidenciado pelo rácio massa da glândula em relação ao peso corporal. Assim, o presente trabalho focou-se nas glândulas submandibulares, que evidenciaram sinais histológicos de hipertrofia das células acinares. A nível molecular, não foram detetadas alterações nos níveis do recetor de androgénio (AR) e do recetor de estrogénio alfa, apesar do aumento significativo dos níveis de hormonas sexuais em circulação. A análise GeLC-MS/MS dos extratos das glândulas submandibulares permitiu identificar 308 proteínas diferentes, 12 das quais foram apenas identificadas no grupo dos animais tratados com testosterona (TEST). Das proteínas comuns identificadas nos grupos controlo (CTRL) e TEST, 14 foram reguladas positivamente pela testosterona, embora não tenham sido observadas interações proteínaproteína significativas entre elas. Para além disso, a análise da atividade proteolítica das glândulas submandibulares por zimografia não revelou diferenças de atividade entre os grupos CTRL e TEST. No entanto, estes resultados destacaram a contribuição das calicreínas (Klk) para a atividade proteolítica destas glândulas. A análise das proteínas reguladas positivamente no grupo TEST evidenciaram uma expressão de Klk3 mais significativa nas glândulas mandibulares dos ratos TEST (aumento de 2 vezes em comparação com o grupo CTRL). Além disso, esta protease foi também a única calicreína que evidenciou interação proteína-proteína com o AR, o que sugere que a Klk3 é um alvo deste recetor. Em conclusão, os resultados obtidos na presente dissertação evidenciaram uma regulação diferencial das glândulas salivares à exposição à testosterona e permitiram caracterizar as alterações do proteoma das glândulas submandibulares em resposta a esta exposição. A compreensão das adaptações moleculares das glândulas salivares pode contribuir para uma melhor compreensão dos mecanismos que regulam a saúde oral e para o desenvolvimento de potenciais intervenções terapêuticas para a sua promoção.2025-07-18T00:00:00Z2023-07-05T00:00:00Z2023-07-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/39618engSantos, João Miguel Valenteinfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T12:17:26Zoai:ria.ua.pt:10773/39618Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:09:46.656594Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv The impact of testosterone on aging-related remodeling of salivary glands
title The impact of testosterone on aging-related remodeling of salivary glands
spellingShingle The impact of testosterone on aging-related remodeling of salivary glands
Santos, João Miguel Valente
Salivary glands
Submandibular gland’s proteome
Testosterone
Aging
Rats
GeLC-MS/MS
title_short The impact of testosterone on aging-related remodeling of salivary glands
title_full The impact of testosterone on aging-related remodeling of salivary glands
title_fullStr The impact of testosterone on aging-related remodeling of salivary glands
title_full_unstemmed The impact of testosterone on aging-related remodeling of salivary glands
title_sort The impact of testosterone on aging-related remodeling of salivary glands
author Santos, João Miguel Valente
author_facet Santos, João Miguel Valente
author_role author
dc.contributor.author.fl_str_mv Santos, João Miguel Valente
dc.subject.por.fl_str_mv Salivary glands
Submandibular gland’s proteome
Testosterone
Aging
Rats
GeLC-MS/MS
topic Salivary glands
Submandibular gland’s proteome
Testosterone
Aging
Rats
GeLC-MS/MS
description Salivary glands play an important role in the secretion of saliva into the oral cavity, influencing salivary composition and oral health. However, the molecular profile of salivary glands is poorly understood, especially in clinical scenarios associated with changes in sex hormone levels. Therefore, our study aimed to investigate how chronic exposure to supraphysiological testosterone levels affects salivary gland remodelling, with a particular focus on proteome adaptation. To achieve this goal, male Wistar rats were implanted subcutaneously at 12 weeks-of-age with devices that release testosterone. At 35 and 61 weeks-of-age, the animals were sacrificed, and their salivary glands were harvested for histological and molecular characterization. The data showed that the submandibular glands were more sensitive to testosterone compared with the parotid glands, as evidenced primarily by a significantly higher ratio of gland mass to body weight. Further analysis of the submandibular glands revealed histologic evidence of acinar cell hypertrophy. At the molecular level, no changes in tissue levels of androgen receptor (AR) and estrogen receptor alpha were detected despite significantly increased levels of circulating sex hormones. GeLC- MS /MS analysis of submandibular gland extracts identified 308 different proteins, 12 of which were modulated by testosterone. Of the common proteins identified from the CTRL and TEST groups, 14 were up-regulated by testosterone, but no relevant interactions were observed between them. In addition, zymographic analysis of the submandibular glands revealed a similar proteolytic profile between the CTRL and TEST groups, with no significant differences observed in optical density values between the two groups. However, our data highlight the contribution of kallikreins (Klk) to the proteolytic activity of these glands, as seen in the zymography gel profile. When looking at the individual proteins that were up-regulated in the TEST group, the stronger expression of Klk3 in the mandibular glands of TEST rats was striking (2-fold increase compared with the CTRL group). Moreover, this protease was also the only kallikrein that showed interaction with the AR, suggesting that Klk3 is a target of this receptor. These results shed light on the differential response of salivary glands to testosterone exposure and provide insight into the proteome-level changes associated with this exposure. Understanding the molecular adaptations in the salivary glands may contribute to a better understanding of oral health and the development of potential therapeutic interventions.
publishDate 2023
dc.date.none.fl_str_mv 2023-07-05T00:00:00Z
2023-07-05
2025-07-18T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10773/39618
url http://hdl.handle.net/10773/39618
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/embargoedAccess
eu_rights_str_mv embargoedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799137747895582720