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TNFα and leptin inhibit basal and glucose-stimulated insulin secretion and gene transcription in the HIT-T15 pancreatic cells

International Journal of Obesity volume 25, pages 1018–1026 (2001)Cite this article

Abstract

BACKGROUND: Tumor necrosis factor α (TNFα), a cytokine produced at inflammatory sites and in adipose tissue, is known primarily for its detrimental effects on insulin action. There is evidence to suggest that TNFα may also influence β-cell function. Leptin is another adipose tissue-derived hormone that might also act on β-cells.

OBJECTIVE: We explored the independent and combined effects of TNFα and leptin upon basal and glucose-stimulated insulin transcription and secretion in the HIT-T15 pancreatic β cell line.

METHODS: Cells were cultured for 40 h in the presence of near-normal basal (7 mM) or high (16.7 mM) glucose and treated with either TNFα (1, 10 and 50 ng/ml) or leptin (10, 50 and 100 ng/ml) or both together. Insulin concentrations were measured by radioimmunoassay. Insulin mRNA levels were evaluated by a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method, after normalization with β-actin mRNA.

RESULTS: TNFα significantly suppressed basal and glucose-stimulated insulin secretion and proinsulin mRNA transcription in a dose-dependent manner, an effect that was more powerful in the presence of high glucose. Leptin also inhibited dose-dependent insulin mRNA and protein at both glucose concentrations, but did not appear to further potentiate the suppressive effects of TNFα.

CONCLUSION: TNFα suppresses both basal and glucose-stimulated insulin transcription and secretion in HIT-T15 cells, an effect that is enhanced significantly by high glucose. Leptin also independently inhibits basal and glucose-stimulated insulin secretion and transcription but does not modify TNFα effects. These effects might contribute to the abnormalities of glucose metabolism that characterize conditions of increased TNFα and/or leptin production.

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Acknowledgements

We thank Mrs Athina Koukourava for her valuable assistance in the insulin secretion measurements.

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  1. Hellenic National Diabetes Centre (HNDC), Athens, Greece

    PC Tsiotra, C Tsigos & SA Raptis

  2. 2nd Propedeutic Department of Internal Medicine, Research Institute and Diabetes Center, University of Athens, Athens, Greece

    SA Raptis

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  1. PC Tsiotra
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Correspondence to C Tsigos.

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Tsiotra, P., Tsigos, C. & Raptis, S. TNFα and leptin inhibit basal and glucose-stimulated insulin secretion and gene transcription in the HIT-T15 pancreatic cells. Int J Obes 25, 1018–1026 (2001). https://doi.org/10.1038/sj.ijo.0801657

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