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4-phenylbutyric acid attenuates endoplasmic reticulum stress-mediated apoptosis and protects the hepatocytes from intermittent hypoxia-induced injury

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Abstract

Purpose

To investigate the effect of 4-phenylbutyric acid (4-PBA) on intermittent hypoxia (IH)-induced liver cell injury and to clarify the underlying mechanisms.

Methods

L02 cells (normal human liver cells) were cultured in normoxic condition or subjected to intermittent hypoxia for 4, 8, and 12 h. A part of hypoxia-treated L02 cells was applied with 4-PBA 1 h before exposure to hypoxia. The effect of 4-PBA on liver injury, hepatocyte apoptosis, endoplasmic reticulum stress (ERS), and PERK-eIFa2-ATF4-CHOP apoptotic pathway was investigated.

Results

(1) IH caused apoptosis in hepatocyte; (2) IH caused ERS in hepatocyte; (3) IH caused hepatic injury; (4) 4-PBA attenuated IH-induced liver cell injury; (5) 4-PBA protected liver cell from IH-induced apoptosis; (6) 4-PBA suppressed ERS-related apoptotic pathway (PERK-eIFa2-ATF4-CHOP), but did not suppress IH-induced unfold protein reaction (UPR).

Conclusions

4-PBA could protect liver cells by suppressing IH-induced apoptosis mediated by ERS, but not by reducing the UPR.

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Abbreviations

4-PBA:

4-phenylbutyric acid

ALT:

alanine aminotransferase

AST:

aspartate aminotransferase

ATF4:

activating transcription Factor 4

CHOP:

C/EBP homologous protein

ERS:

endoplasmic reticulum stress

FBS:

fetal bovine serum

GRP78:

glucose-regulated protein 78

IH:

intermittent hypoxia

NC:

negative control

OSA:

obstructive sleep apnea

PBS:

phosphate-buffered saline

p-eIF2α:

phosphorylated-eukaryotic translation initiation factor 2 subunit α

PMSF:

phenylmethanesulphonylfluoride

p-PERK:

phosphorylated-PKR-like ER protein kinase

UPR:

unfold protein reaction

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Funding

National Natural Science Foundation Committee of China provided financial support in the form of National Natural Science Foundation of China funding (No. 81670086, No. 81370183). Natural Science Foundation Committee of Tianjin City provided financial support in the form of Natural Science Foundation of Tianjin City funding (No. 14JCYBJC27800) and major special project for the prevention and control of chronic diseases in Tianjin (Grant No. 17ZXMFSY00080). The sponsor had no role in the design or conduct of this research.

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Authors and Affiliations

  1. Geriatrics, Institute of Gerontology of Tianjin, Tianjin Medical University General Hospital, No.154, Anshan Road, Heping District, Tianjin, China

    Liu Xin, Wu Fan, Du Tingting & Zhang Qiang

  2. Beckman Research Institute of the City of Hope, Duarte, CA, USA

    Sun Zuoming

Authors
  1. Liu Xin
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  2. Wu Fan
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  3. Du Tingting
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  4. Sun Zuoming
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  5. Zhang Qiang
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Correspondence to Zhang Qiang.

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The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Comment

This study shows that increasing exposure of hepatocytes to hypoxia demonstrates hypoxia causes a dose-dependent injury which is partly attenuated by 4-phenylbutyric acid. The role of intermittent hypoxia in the pathophysiology of OSA-related liver dysfunction is clearly of interest to scientists and clinicians alike; this study helps advance our understanding of this area and potential therapeutic pathways.

While this is an in vitro study, it does provide support for the notion that currently accepted adequate notions of treatment efficacy may need to be reconsidered. Perhaps with CPAP, some is helpful, more is better, and most of the night is better still.

Ian Wilcox

NSW, Australia

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Xin, L., Fan, W., Tingting, D. et al. 4-phenylbutyric acid attenuates endoplasmic reticulum stress-mediated apoptosis and protects the hepatocytes from intermittent hypoxia-induced injury. Sleep Breath 23, 711–717 (2019). https://doi.org/10.1007/s11325-018-1739-y

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