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Reduced endoplasmic reticulum luminal calcium links saturated fatty acid-mediated endoplasmic reticulum stress and cell death in liver cells

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An Erratum to this article was published on 24 October 2009

An Erratum to this article was published on 11 July 2009

Abstract

Chronic exposure to elevated free fatty acids, in particular long chain saturated fatty acids, provokes endoplasmic reticulum (ER) stress and cell death in a number of cell types. The perturbations to the ER that instigate ER stress and activation of the unfolded protein in response to fatty acids in hepatocytes have not been identified. The present study employed H4IIE liver cells and primary rat hepatocytes to examine the hypothesis that saturated fatty acids induce ER stress via effects on ER luminal calcium stores. Exposure of H4IIE liver cells and primary hepatocytes to palmitate and stearate reduced thapsigargin-sensitive calcium stores and increased biochemical markers of ER stress over similar time courses (6 h). These changes preceded cell death, which was only observed at later time points (16 h). Co-incubation with oleate prevented the reduction in calcium stores, induction of ER stress markers and cell death observed in response to palmitate. Inclusion of calcium chelators, BAPTA-AM or EGTA, reduced palmitate- and stearate-mediated enrichment of cytochrome c in post-mitochondrial supernatant fractions and cell death. These data suggest that redistribution of ER luminal calcium contributes to long chain saturated fatty acid-mediated ER stress and cell death.

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Acknowledgments

This work was supported by grants DK47416 and DK072017 from the National Institutes of Health and the Lillian Fountain Smith Foundation Endowment.

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Authors and Affiliations

  1. Department of Food Science and Human Nutrition, Colorado State University, Gifford 234, Fort Collins, CO, 80523-1571, USA

    Yuren Wei, Dong Wang, Christopher L. Gentile & Michael J. Pagliassotti

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  1. Yuren Wei
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  2. Dong Wang
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Correspondence to Michael J. Pagliassotti.

Additional information

An erratum to this article can be found at http://dx.doi.org/10.1007/s11010-009-0302-3

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Supplemental Fig. 1

(A) Caspase-3 activity in H4IIE liver cells or primary hepatocytes incubated for 6 or 16 h in control media (LG), or control media supplemented with thapsigargin (Thap, 450 nM), oleate (O, 250 μM), or palmitate (P, 250 μM). (B) Caspase-3 activity in H4IIE liver cells or primary rat hepatocytes incubated for 16 h in control media (LG), or control media supplemented with palmitate (P, 250 μM) or palmitate (P, 250 μM) + oleate (O, 125 μM). (C) Caspase-3 activity in H4IIE liver cells or primary rat hepatocytes incubated for 16 h in control media (LG), or control media supplemented with oleate (O, 250 μM), linoleate (L, 250 μM), palmitate (P, 250 μM) or stearate (S, 250 μM) in the absence (no additions) or presence of BAPTA-AM (20 μM) or EGTA (1 mM). Data are reported as the mean ± SD for triplicate samples from 4 to 6 independent experiments. *Significantly (P < 0.05) different from LG−. +Significantly (P < 0.05) different from no additions of the same treatment group

Supplemental Fig. 2

(A) Real time PCR analysis of CHOP, GADD34, ATF4 mRNA in H4IIE liver cells and primary hepatocytes following 16 h incubations in control media (LG), or control media supplemented with oleate (O, 250 μM), linoleate (L, 250 μM), palmitate (P, 250 μM) or stearate (S, 250 μM) in the absence (no additions) or presence of BAPTA-AM (20 μM) or EGTA (1 mM). LG with no additions was set to 1. (B) MTT cell viability assay following 16 h incubations. Data are expressed as the mean ± SD for 4–6 independent experiments

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Wei, Y., Wang, D., Gentile, C.L. et al. Reduced endoplasmic reticulum luminal calcium links saturated fatty acid-mediated endoplasmic reticulum stress and cell death in liver cells. Mol Cell Biochem 331, 31–40 (2009). https://doi.org/10.1007/s11010-009-0142-1

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