Short-Term Protocols to Obtain Insulin-Producing Cells from Rat Adipose Tissue: Signaling Pathways and In Vivo Effect.

Studies using mesenchymal stromal cells (MSCs) as a source of insulin-secreting cells (IPCs) are a promising path in the pursuit for diabetes therapy. Here, we investigate three short-term differentiation protocols in order to generate IPCs from autologous adipose-derived stromal cells (ADSCs) with an expressive insulin-secreting profile in vitro and in vivo, as well as the signaling pathways involved in the chosen differentiation protocols. We extracted and cultured ADSCs and differentiated them into IPCs, using three different protocols with different inductors. Afterwards, the secretory profile was analyzed and IPCs differentiated in exendin-4/activin A medium, which presented the best secretory profile, was implanted in the kidney subcapsular region of diabetic rats. All protocols induced the differentiation, but media supplemented with exendin-4/activin A or resveratrol induced the expression and secretion of insulin more efficiently, and only the exendin-4/activin-A-supplemented medium generated an insulin secretion profile more like ß-cells, in response to glucose. The PI3K/Akt pathway seems to play a negative role in IPC differentiation; however, the differentiation of ADSCs with exendin-4/activin A positively modulated the p38/MAPK pathway. Resveratrol medium activated the Jak/STAT3 pathway and generated IPCs apparently less sensitive to insulin and insulin-like receptors. Finally, the implant of IPCs with the best secretory behavior caused a decrease in hyperglycemia after one-week implantation in diabetic rats. Our data provide further information regarding the generation of IPCs from ADSCs and strengthen evidence to support the use of MSCs in regenerative medicine, specially the use of exendin-4/activin A to produce rapid and effectively IPCs with significant in vivo effects.

Streptozotocin causes acute responses on hippocampal S100B and BDNF proteins linked to glucose metabolism alterations.

Streptozotocin (STZ) is a glucosamine-nitrosourea commonly used to induce long-lasting models of diabetes mellitus and Alzheimer's disease. Direct toxicity of STZ on the pancreas and kidneys has been well characterized, but the acute effect of this compound on brain tissue has received less attention. Herein, we investigated the acute and direct toxicity of STZ on fresh hippocampal slices, measuring changes in BDNF and S100B secretion (two widely-used peripheral markers of brain injury), as well as glucose metabolism. Moreover, we investigated in vivo changes of these proteins in the hippocampus, 48 h after intracerebroventricular STZ administration. Transverse hippocampal slices (0.3 mm thick) were obtained using a McIlwain tissue chopper and target proteins were measured in the incubation medium by ELISA. STZ decreased S100B secretion, but increased BDNF secretion as well as causing impairment in glucose uptake in hippocampal slices, measured using [3H] deoxy-glucose. Glucose levels and glucose metabolism differentially modulated S100B secretion in astrocytes and BDNF secretion in neurons, when evaluated under specific conditions (high-potassium medium, presence of tetrodotoxin or fluorocitrate). Moreover, at 48 h after intracerebroventricular STZ, hippocampal BDNF content, but not S100B, was reduced. Our results indicate that BDNF and S100B are useful and sensitive markers of glucose metabolism disturbance and reinforce these proteins as general acute markers of brain disorders.

Hippocampal changes in STZ-model of Alzheimer's disease are dependent on sex.

The majority of Alzheimer's disease (AD) cases are sporadic and aging is the major risk factor for developing the disease, affecting more women than men. In spite of different gender prevalence, most experimental studies in animal models have been performed in male. This study investigates the streptozotocin (STZ)-induced AD model at three different times (2, 4 and 8 weeks afterwards) and in male and female rats, evaluating cognitive deficit, cholinergic neurotransmission, glucose uptake, glutathione content and specific glial markers (GFAP and S100B protein) in the hippocampus of the rat. Our data reinforce the relevance of alterations in STZ model of dementia, reported in the genesis and/or progression of AD such as cholinergic deficit and glucose uptake decrease. All alterations in these parameters (except GFAP) were dependent on sex. It is unclear, at this moment, which alterations are due to sex steroid modulation. In spite of limitations of this experimental model, these data may contribute to understand AD susceptibility and progression dependent on sex.