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Long-term exposure of INS-1 rat insulinoma cells to linoleic acid and glucose in vitro affects cell viability and function through mitochondrial-mediated pathways

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Abstract

Obesity with excessive levels of circulating free fatty acids (FFAs) is tightly linked to the incidence of type 2 diabetes. Insulin resistance of peripheral tissues and pancreatic β-cell dysfunction are two major pathological changes in diabetes and both are facilitated by excessive levels of FFAs and/or glucose. To gain insight into the mitochondrial-mediated mechanisms by which long-term exposure of INS-1 cells to excess FFAs causes β-cell dysfunction, the effects of the unsaturated FFA linoleic acid (C 18:2, n-6) on rat insulinoma INS-1 β cells was investigated. INS-1 cells were incubated with 0, 50, 250 or 500 μM linoleic acid/0.5% (w/v) BSA for 48 h under culture conditions of normal (11.1 mM) or high (25 mM) glucose in serum-free RPMI-1640 medium. Cell viability, apoptosis, glucose-stimulated insulin secretion, Bcl-2, and Bax gene expression levels, mitochondrial membrane potential and cytochrome c release were examined. Linoleic acid 500 μM significantly suppressed cell viability and induced apoptosis when administered in 11.1 and 25 mM glucose culture medium. Compared with control, linoleic acid 500 μM significantly increased Bax expression in 25 mM glucose culture medium but not in 11.1 mM glucose culture medium. Linoleic acid also dose-dependently reduced mitochondrial membrane potential (ΔΨm) and significantly promoted cytochrome c release from mitochondria in both 11.1 mM glucose and 25 mM glucose culture medium, further reducing glucose-stimulated insulin secretion, which is dependent on normal mitochondrial function. With the increase in glucose levels in culture medium, INS-1 β-cell insulin secretion function was deteriorated further. The results of this study indicate that chronic exposure to linoleic acid-induced β-cell dysfunction and apoptosis, which involved a mitochondrial-mediated signal pathway, and increased glucose levels enhanced linoleic acid-induced β-cell dysfunction.

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Acknowledgments

The authors would like to acknowledge scientific contribution of Drs SH Lee and F Steyn, and editorial help of Mr M Sinclair. Ya Tuo is a recipient of overseas post-graduate research scholarship from the China Scholarship Council (CSC, China) and subsidy scholarship by the University of Queensland; and research was supported by Australian NHMRC and the University of Queensland

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Authors and Affiliations

  1. Department of Forensic Science, School of Medicine, Xi’an Jiaotong University, Xi’an, Shaanxi, China

    Ya Tuo & Shengbin Li

  2. School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, 4072, Australia

    Ya Tuo, Dengfeng Wang & Chen Chen

  3. Department of Gynaecology and Obstetrics, West China Secondary Hospital, Sichuan University, Chengdu, China

    Dengfeng Wang

  4. Research Institute of Metabolic Disorders (RIMD), Xi’an Jiaotong University, Xi’an, Shaanxi, China

    Chen Chen

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  1. Ya Tuo
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Correspondence to Chen Chen.

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Shengbin Li and Chen Chen contributed equally to this work.

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Tuo, Y., Wang, D., Li, S. et al. Long-term exposure of INS-1 rat insulinoma cells to linoleic acid and glucose in vitro affects cell viability and function through mitochondrial-mediated pathways. Endocr 39, 128–138 (2011). https://doi.org/10.1007/s12020-010-9432-3

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