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Control of ER stress by a chemical chaperone counteracts apoptotic signals in IFN-γ-treated murine hepatocytes

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Abstract

Apoptosis of hepatocytes plays a key role in the pathogenesis of immune-mediated hepatitis. However, the detailed mechanisms of apoptotic signaling remain unclear. In this study, we investigated the involvement of ER stress in a model of IFN-γ-induced apoptosis of hepatocytes in vitro, using a chemical chaperone reagent, glycerol. IFN-γ-induced apoptotic events (mitochondrial release of cytochrome c, enzymatic activation of caspase-3 and -9) were markedly inhibited by glycerol. Glycerol induced partial inhibition of cytotoxicity indicated by lactate dehydrogenase release from the cytosol but had no inhibitory effect on the induction of IRF-1 gene expression and reactive oxygen species, required for hepatocyte apoptosis by IFN-γ. Induction of caspase-4 and -12 gene expression, positively correlated with ER stress, was attenuated by glycerol. Gene analysis revealed that induction of ER stress-related genes, C/EBP homologue protein (CHOP/GADD153) and TRB3, was suppressed completely by glycerol treatment. These results suggest that ER stress plays a crucial role in mediating apoptosis of hepatocytes induced by IFN-γ, and a chemical chaperone is an effective inhibitor of the ER stress.

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Abbreviations

AG:

Aminoguanidine

CHOP:

C/EBP homologue protein

ER:

Endoplasmic reticulum

IFN-γ:

Interferon gamma

LDH:

Lactate dehydrogenase

4-PBA:

Sodium 4-phenylbutyrate

PDTC:

Pyrrolidinedithiocarbamate

ROS:

Reactive oxygen species

SERCA:

Sarcoplasmic/endoplasmic Ca2+-ATPase

TG:

Thapsigargin

UPR:

Unfold protein response

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Acknowledgments

This work was supported by the High-Tech Research Center Project for Private Universities: matching fund subsidy from MEXT (Ministry of Education, Culture, Sports, Science and Technology), 2004–2009.

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  1. Department of Pharmaceutical Sciences, Musashino University, 1-1-20 Shinmachi, Nishitokyo, Tokyo, 202-8585, Japan

    Keita Kanki, Takeshi Kawamura & Yoshifumi Watanabe

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  1. Keita Kanki
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  2. Takeshi Kawamura
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  3. Yoshifumi Watanabe
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Correspondence to Yoshifumi Watanabe.

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Kanki, K., Kawamura, T. & Watanabe, Y. Control of ER stress by a chemical chaperone counteracts apoptotic signals in IFN-γ-treated murine hepatocytes. Apoptosis 14, 309–319 (2009). https://doi.org/10.1007/s10495-009-0318-x

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