Nuclear factor-κB and advanced glycation end-products expression in lacrimal glands of aging rats

Alves, Mônica; Cunha, Daniel Andrade Da; Calegari, C.; Saad, Mário J.A.; Boschero, Carlos A.; Velloso, Lı́cio A.; Melani, Ela

doi:10.1530/joe-17-6209x

Cited by 2 publications

(3 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This protein drives vesicular docking and apical membrane fusion and it can also play a role in de termining vesicular size (17) (Figure 1). In previous studies, aging rats had presented with dry eye disease, an insulin-signaling im pairment, oxidative stress, and declines in the parasympathetic control of LG function; this revealed an interesting similarity with DM dry eye (18,19) . Some SNARE and Rab family members were also altered in aged LG; particularly, Rab 3 expression was reduced at both the mRNA and protein levels (20) .…”

Section: Insulin Replacement Restores the Vesicular Secretory Apparatmentioning

confidence: 85%

See 1 more Smart Citation

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

Dias

Batista²,

Roma³

et al. 2015

Arquivos Brasileiros De Oftalmologia

View full text Add to dashboard Cite

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

show abstract

Section: Insulin Replacement Restores the Vesicular Secretory Apparatmentioning

confidence: 85%

“…Some SNARE and Rab family members were also altered in aged LG; particularly, Rab 3 expression was reduced at both the mRNA and protein levels (20) . In addition, advanced glycation end pro ducts can accumulate in aged and diabetic rat LGs (7,18) .…”

Section: Insulin Replacement Restores the Vesicular Secretory Apparatmentioning

confidence: 99%

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

Dias

Batista²,

Roma³

et al. 2015

Arquivos Brasileiros De Oftalmologia

View full text Add to dashboard Cite

show abstract

“…An increase in both the number of living cells and cell viability with insulin at 5.0 μg/mL was observed from the third to the ninth day of cultivation, compared with day 0 (seeding), as well as with lower or higher doses of insulin, indicating a key role of insulin in a certain range for the homeostasis and proliferation of LGAC (8,20) . The trend to form clusters after the third day revealed that these cells show polarity, as previously observed (15) .…”

mentioning

confidence: 99%

Lacrimal gland primary acinar cell culture: the role of insulin

Malki

Dias

Jorge

et al. 2016

Arquivos Brasileiros De Oftalmologia

View full text Add to dashboard Cite

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

show abstract

Nuclear factor-κB and advanced glycation end-products expression in lacrimal glands of aging rats

Cited by 2 publications

References 24 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

Lacrimal gland primary acinar cell culture: the role of insulin

Contact Info

Product

Resources

About