Influence of insulin treatment on the lacrimal gland and ocular surface of diabetic rats

Módulo, Carolina Maria; Jorge, A. G.; Dias, Ana Carolina; Braz, Alexandre Martins; Bertazolli-Filho, Rubens; Jordão, Alceu Afonso; Marchini, Júlio Sérgio; Rocha, Eduardo Melani

doi:10.1007/s12020-009-9208-9

Cited by 42 publications

(48 citation statements)

References 51 publications

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“…Such an insight will provide a better understanding of dry eye pathophysiology (7,12,21) . The major current finding was that lower intracellular transport protein expression levels in the DM group were associated with declines in delimited SV density and their ultra-structural appearance of the acinar apical cell membranes.…”

Section: Discussionmentioning

confidence: 99%

“…In an aging rodent, which is another model of the dry eye syndrome, similar alterations in SV appearance and declines in their density were associated with lower Rab 3d levels (20) . Similar to the DM model used here, this model had high levels of oxidative stress and insulin resistance (19,21) . As in that study, we were unable to identify syntaxin 1 at the mRNA levels, but detected it with Western blotting.…”

Section: Discussionmentioning

confidence: 99%

“…This dose was not sufficient to maintain adequate glycemic control, but was enough to avoid body weight loss and growth retardation (21) . Comparative studies of the three groups, i.e., control (C), diabetic (DM), and diabetic with insulin treatment (IT), were performed 10 weeks later.…”

Section: Animal Modelmentioning

confidence: 99%

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Insulin replacement restores the vesicular secretory apparatus in the diabetic rat lacrimal gland

Dias

Batista²,

Roma³

et al. 2015

Arquivos Brasileiros De Oftalmologia

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Purpose: In the lacrimal gland (LG) acinar cells, signaling regulates the release of secretory vesicles through specific Rab and SNARE exocytotic proteins. In diabetes mellitus (DM), the LGs are dysfunctional. The aim of this work was to determine if secretory apparatus changes were associated with any effects on the secretory vesicles (SV) in diabetic rats as well as the expression levels of constituent Rab and members of the SNARE family, and if insulin supplementation reversed those changes. Methods: DM was induced in male Wistar rats with an intravenous dose of streptozotocin (60 mg/kg). One of the two diabetic groups was then treated every other day with insulin (1 IU). A third control group was injected with vehicle. After 10 weeks, Western blotting and RT-PCR were used to compared the Rab and SNARE secretory factor levels in the LGs. Transmission electron microscopy evaluated acinar cell SV density and integrity. Results: In the diabetes mellitus group, there were fewer and enlarged SV. The Rab 27b, Rab 3d, and syntaxin-1 protein expression declined in the rats with diabetes mellitus. Insulin treatment restored the SV density and the Rab 27b and syntaxin expression to their control protein levels, whereas the Vamp 2 mRNA expression increased above the control levels. Conclusions: Diabetes mellitus LG changes were associated with the declines in protein expression levels that were involved in supporting exocytosis and vesicular formation. They were partially reversed by insulin replacement therapy. These findings may help to improve therapeutic management of dry eye in diabetes mellitus.Keywords: Diabetes mellitus/chemically induced; Lacrimal apparatus; Exocytosis; secretory vesicles; R-SNARE proteins; Animals; Rats RESUMO Objetivo: Células acinares da glândula lacrimal (GL) sinalizam a regulação da liberação através de vesículas secretórias específicas Rab proteínas exocitóticas SNARE. No diabetes mellitus (DM), as glândulas lacrimais são disfuncionais. O objetivo deste trabalho foi determinar se em ratos diabéticos, alterações dos aparatos secretórios estão associados a efeitos sobre vesículas secretoras (VS) e sobre os níveis de expressão do constituinte Rab, bem como membros da família SNARE, e se a suplementação de insulina reverte as alterações. Métodos: DM foi induzido em ratos Wistar machos com uma dose intravenosa de estreptozotocina (60 mg/kg). Um dos dois grupos diabéticos foi então tratado a cada dois dias com insulina (1 UI). Um terceiro grupo controle foi injetado com o veículo. Após 10 semanas, western blot e RT-PCR comparou níveis de fatores secretórios de

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Insulin replacement restores the vesicular secretory apparatus in the diabetic rat lacrimal gland

Dias

Batista²,

Roma³

et al. 2015

Arquivos Brasileiros De Oftalmologia

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“…A lesão celular causada pelos radicais livres é resultante de peroxidação lipídica das membranas, modificação oxidativa de proteínas e dano oxidativo ao DNA (11) . Há ainda indícios de que o estresse oxidativo relacionado ao estado de hiperglicemia em casos de diabetes mellitus e resistência à insulina esteja envolvido na produção das alterações histológicas encontradas na glândula lacrimal de ratos, caracterizadas por mudanças na morfologia e acúmulo de inclusões semelhantes a lipofuscina (12) . As variações fisiológicas que se desenvolvem com o envelhecimento também estão envolvidas na gênese do olho seco e incluem diminuição de volume e fluxo lacrimal, aumento da osmolaridade, perda da estabilidade do filme lacrimal e alterações na composição lipídica das glândulas de Meibômio.…”

Section: Fisiopatogeniaunclassified

Olho seco: etiopatogenia e tratamento

Fonseca¹,

Arruda²,

Rocha³

2010

Arq. Bras. Oftalmol.

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“…Insulin is thought to be of major importance, based on observations related to its deprivation in diabetes mellitus (DM), on LG, tear film, and ocular surface in humans, animal models, and in culture (6)(7)(8) . Lacrimal gland acinar cell (LGAC) culture is an appropriate model for improving the understanding of the influence of insulin on these cells.…”

mentioning

confidence: 99%

Lacrimal gland primary acinar cell culture: the role of insulin

Malki

Dias

Jorge

et al. 2016

Arquivos Brasileiros De Oftalmologia

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The condition of lacrimal glands (LG) is influenced by components of the immune, neural, and endocrine systems (1)(2)(3)(4)(5) . Insulin is thought to be of major importance, based on observations related to its deprivation in diabetes mellitus (DM), on LG, tear film, and ocular surface in humans, animal models, and in culture (6)(7)(8) . Lacrimal gland acinar cell (LGAC) culture is an appropriate model for improving the understanding of the influence of insulin on these cells.In previous studies, the culture of LGACs from different species was performed over an average period of 21 days (9,10) . Several attempts to enhance the growth of acinar cells in culture have been made, but after a span of approximately 3 weeks, LGACs exhibited decreased viability, with high numbers of apoptotic cells (11)(12)(13)(14) . Exocrine acinar cells are fragile, post-mitotic epithelial cells with marked polarity. This indicates that the apical and basal sides are clearly positioned but also that intracellular organelles are located at one or the other pole, depending on their cellular functions. Thus, an adequate extracellular matrix and other specific ingredients are needed in the culture media to mimic in vivo conditions (15,16) . The handling during isolation and culture procedure of LGACs should be gentle ABSTRACT Purpose: The goal of the present study was to establish a protocol for primary culture of lacrimal gland acinar cells (LGACs) and to assess the effect of adding insulin to the culture media. Methods:LGACs were isolated and cultured from lacrimal glands of Wistar male rats. The study outcomes included cell number, viability, and peroxidase release over time and in response to three concentrations of insulin (0.5, 5.0, and 50.0 μg/mL). Results: In LGAC primary culture, cells started to form clusters by day 3. There was a time-response pattern of peroxidase release, which rose by day 6, in response to carbachol. Culture viability lasted for 12 days. An insulin concentration of 5.0 μg/mL in the culture medium resulted in higher viability and secretory capacity. Conclusions:The present method simplifies the isolation and culture of LGACs. The data confirmed the relevance of adding insulin to maintain LGACs in culture. Keywords

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Influence of insulin treatment on the lacrimal gland and ocular surface of diabetic rats

Cited by 42 publications

References 51 publications

Insulin replacement restores the vesicular secretory apparatus in the diabetic rat lacrimal gland

Insulin replacement restores the vesicular secretory apparatus in the diabetic rat lacrimal gland

Olho seco: etiopatogenia e tratamento

Lacrimal gland primary acinar cell culture: the role of insulin

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