Effects of Prolonged Exposure in Vitro to Interferon-γ and Tumour Necrosis Factor-α on Nitric Oxide and Insulin Production of Rat Pancreatic Islets

Abstract

It has been postulated that cytokines may mediate the β-cell destructive process causing insulin-dependent diabetes mellitus. The aim of this investigation was to study cytokine effects on pancreatic islet functions in vitro. For this purpose 5–7 days precultured (medium RPMI 1640+ 10% fetal calf serum) rat pancreatic islets were exposed for another 48 h to either culture medium alone or with addition of rat interferon-γ (IFN-γ; 1000 U/ml), or human tumor necrosis factor-α (TNF-α; 1000 U/ml) or a combination of the cytokines. After the culture period the islets were subjected to short-term experiments in the absence of cytokines. Neither the DNA nor the insulin content of the islets were affected by the cytokines alone or by the combination. The combination IFN-γ + TNF-α caused a 5-fold increase in the medium nitrite accumulation, indicating induction of nitric oxide formation. It was found that IFN-γ reduced medium insulin accumulation and basal insulin secretion at 1.7 mM glucose, without affecting the medium nitrite level. On the other hand, the islet glucose oxidation rate at 16.7 mM glucose and the insulin secretory response to 16.7 mM glucose was normal or even increased when examined after 48 h. TNF-α alone had no significant effects. In conclusion, a combination of the cytokines can induce nitric oxide formation and inhibition of insulin production in rat pancreatic islets. However, this effect appears not to be sustained. Moreover, IFN-γ alone seems to induce changes not related to nitric oxide.

Key Words:

• β-cells
• Cytokines
• Interferon-γ
• Nitric oxide
• Pancreatic islets
• TNF-α