46 research outputs found
Implanting 1.1B4 human β-cell pseudoislets improves glycaemic control in diabetic severe combined immune deficient mice
AIM: To investigate the potential of implanting pseudoislets formed from human insulin-releasing β-cell lines as an alternative to islet transplantation. METHODS: In this study, the anti-diabetic potential of novel human insulin releasing 1.1B4 β-cells was evaluated by implanting the cells, either as free cell suspensions, or as three-dimensional pseudoislets, into the subscapular region of severe combined immune deficient mice rendered diabetic by single high-dose administration of streptozotocin. Metabolic parameters including food and fluid intake, bodyweight and blood glucose were monitored throughout the study. At the end of the study animals were given an intraperitoneal glucose tolerance test. Animals were then culled and blood and tissues were collected for analysis. Insulin and glucagon contents of plasma and tissues were measured by insulin radioimmunoassay and chemiluminescent enzyme-linked immunosorbance assay respectively. Histological analyses of pancreatic islets were carried out by quantitative fluorescence immunohistochemistry staining. RESULTS: Both pseudoislet and cell suspension implants yielded well vascularised β-cell masses of similar insulin content. This was associated with progressive amelioration of hyperphagia (P < 0.05), polydipsia (P < 0.05), body weight loss (P < 0.05), hypoinsulinaemia (P < 0.05), hyperglycaemia (P < 0.05 - P < 0.001) and glucose tolerance (P < 0.01). Islet morphology was also significantly improved in both groups of transplanted mice, with increased β-cell (P < 0.05 - P < 0.001) and decreased alpha cell (P < 0.05 - P < 0.001) areas. Whereas mice receiving 1.1B4 cell suspensions eventually exhibited hypoglycaemic complications, pseudoislet recipients displayed a more gradual amelioration of diabetes, and achieved stable blood glucose control similar to non-diabetic mice at the end of the study. CONCLUSION: Although further work is needed to address safety issues, these results provide proof of concept for possible therapeutic applicability of human β-cell line pseudoislets in diabetes
Differential Acute and Long Term Actions of Succinic Acid Monomethyl Ester Exposure on Insulin-Secreting BRIN-BD11 Cells
Esters of succinic acid are potent insulin secretagogues,
and have been proposed as novel antidiabetic
agents for type 2 diabetes. This study
examines the effects of acute and chronic exposure
to succinic acid monomethyl ester (SAM) on insulin
secretion, glucose metabolism and pancreatic beta
cell function using the BRIN-BD11 cell line. SAM
stimulated insulin release in a dose-dependent
manner at both non-stimulatory (1.1mM) and stimulatory
(16.7mM) glucose. The depolarizing actions
of arginine also stimulated a significant increase
in SAM-induced insulin release but 2-ketoisocaproic
acid (KIC) inhibited SAM induced insulin
secretion indicating a possible competition between
the preferential oxidative metabolism of these two
agents. Prolonged (18hour) exposure to SAM revealed
decreases in the insulin-secretory responses
to glucose, KIC, glyceraldehyde and alanine.
Furthermore, SAM diminished the effects of nonmetabolized
secretagogues arginine and 3-isobutyl-1-methylxanthine (IBMX). While the ability of
BRIN-BD11 cells to oxidise glucose was unaffected
by SAM culture, glucose utilization was substantially
reduced. Collectively, these data suggest that
while SAM may enhance the secretory potential of
non-metabolized secretagogues, it may also serve as
a preferential metabolic fuel in preference to other
important physiological nutrients and compromise
pancreatic beta cell function following prolonged
exposure
Stem Cell-Based Approaches for the Treatment of Diabetes
The incidence of diabetes and the associated debilitating complications are increasing at an alarming rate worldwide. Current therapies for type 1 diabetes focus primarily on administration of exogenous insulin to help restore glucose homeostasis. However, such treatment rarely prevents the long-term complications of this serious metabolic disorder, including neuropathy, nephropathy, retinopathy, and cardiovascular disease. Whole pancreas or islet transplantations have enjoyed limited success in some individuals, but these approaches are hampered by the shortage of suitable donors and the burden of lifelong immunosuppression. Here, we review current approaches to differentiate nonislet cell types towards an islet-cell phenotype which may be used for larger-scale cell replacement strategies. In particular, the differentiation protocols used to direct embryonic stem cells, progenitor cells of both endocrine and nonendocrine origin, and induced pluripotent stem cells towards an islet-cell phenotype are discussed
L-Arginine Is Essential for Pancreatic b-Cell Functional Integrity, Metabolism and Defense From Inflammatory Challenge
In this work, our aim was to determine whether L-arginine (a known insulinotropic amino acid) can promote a shift of b-cell intermediary metabolism favoring glutathione (GSH) and glutathione disulfide (GSSG) antioxidant responses, stimulus–secretion coupling and functional integrity. Clonal BRIN-BD11 b-cells and mouse islets were cultured for 24 h at various L-arginine concentrations (0–1.15 mmol/l) in the absence or presence of a proinflammatory cytokine cocktail (interleukin 1b, tumour necrosis factor a and interferon g). Cells were assessed for viability, insulin secretion, GSH, GSSG, glutamate, nitric oxide (NO), superoxide, urea, lactate and for the consumption of glucose and glutamine. Protein levels of NO synthase-2, AMP-activated protein kinase (AMPK) and the heat shock protein 72 (HSP72) were also evaluated. We found that L-arginine at 1.15 mmol/l attenuated the loss of b-cell viability observed in the presence of proinflammatory cytokines. L-Arginine increased total cellular GSH and glutamate levels but reduced the GSSG/GSH ratio and glutamate release. The amino acid stimulated glucose consumption in the presence of cytokines while also stimulating AMPK phosphorylation and HSP72 expression. Proinflammatory cytokines reduced, by at least 50%, chronic (24 h) insulin secretion, an effect partially attenuated by L-arginine. Acute insulin secretion was robustly stimulated by L-arginine but this effect was abolished in the presence of cytokines. We conclude that L-arginine can stimulate b-cell insulin secretion, antioxidant and protective responses, enabling increased functional integrity of b-cells and islets in the presence of proinflammatory cytokines. Glucose consumption and intermediary metabolism were increased by L-arginine. These results highlight the importance of L-arginine availability for b-cells during inflammatory challeng
Molecular determinants and intracellular targets of taurine signalling in pancreatic islet β‐cells
AbstractAimDespite its abundance in pancreatic islets of Langerhans and proven antihyperglycemic effects, the impact of the essential amino acid, taurine, on islet β‐cell biology has not yet received due consideration, which prompted the current studies exploring the molecular selectivity of taurine import into β‐cells and its acute and chronic intracellular interactions.MethodsThe molecular aspects of taurine transport were probed by exposing the clonal pancreatic BRIN BD11 β‐cells and primary mouse and human islets to a range of the homologs of the amino acid (assayed at 2–20 mM), using the hormone release and imaging of intracellular signals as surrogate read‐outs. Known secretagogues were employed to profile the interaction of taurine with acute and chronic intracellular signals.ResultsTaurine transporter TauT was expressed in the islet β‐cells, with the transport of taurine and homologs having a weak sulfonate specificity but significant sensitivity to the molecular weight of the transporter. Taurine, hypotaurine, homotaurine, and β‐alanine enhanced insulin secretion in a glucose‐dependent manner, an action potentiated by cytosolic Ca2+ and cAMP. Acute and chronic β‐cell insulinotropic effects of taurine were highly sensitive to co‐agonism with GLP‐1, forskolin, tolbutamide, and membrane depolarization, with an unanticipated indifference to the activation of PKC and CCK8 receptors. Pre‐culturing with GLP‐1 or KATP channel inhibitors sensitized or, respectively, desensitized β‐cells to the acute taurine stimulus.ConclusionTogether, these data demonstrate the pathways whereby taurine exhibits a range of beneficial effects on insulin secretion and β‐cell function, consistent with the antidiabetic potential of its dietary low‐dose supplementation