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Endoplasmic Reticulum Stress-Activated Glycogen Synthase Kinase 3β Aggravates Liver Inflammation and Hepatotoxicity in Mice with Acute Liver Failure

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Abstract

Endoplasmic reticulum stress (ER stress) has been increasingly recognized as an important mechanism in various liver diseases. However, its intrinsic physiological role in acute liver failure (ALF) remains largely undetermined. This study aimed to examine how ER stress orchestrates glycogen synthase kinase 3β (GSK3β) and inflammation to affect ALF. In a murine ALF model induced by d-galactosamine (d-GalN) and lipopolysaccharide (LPS), 4-phenylbutyric acid (4-PBA) is to be administered to relieve ER stress. The lethality rate, liver damage, cytokine expression, and the activity of GSK3β were evaluated. How to regulate LPS-induced inflammation and TNF-α-induced hepatocyte apoptosis by ER stress was investigated in vitro. In vivo, ER stress was triggered in the liver with the progression of mice ALF model. ER stress was essential for the development of ALF because ER stress inhibition by 4-PBA ameliorated the liver damage through decreasing liver inflammation and hepatocyte apoptosis. 4-PBA also decreased GSK3β activity in the livers of ALF mice. In vitro, ER stress induced by tunicamycin synergistically increased LPS-triggered pro-inflammatory cytokine induction and promoted the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway in bone marrow-derived macrophages; moreover, tunicamycin also cooperated with TNF-α to increase hepatocyte apoptosis. ER stress promoted LPS-triggered inflammation depending on GSK3β activation because inhibition of GSK3β by SB216763, the specific inhibitor of GSK3β, resulted in downregulation of pro-inflammatory genes. ER stress contributes to liver inflammation and hepatotoxicity in ALF, particularly by regulating GSK3β, and is therefore a potential therapeutic target for ALF.

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Abbreviations

ALF:

Acute liver failure

ER:

Endoplasmic reticulum

ER stress:

Endoplasmic reticulum stress

GSK3β:

Glycogen synthase kinase 3β

d-GalN:

d-galactosamine

LPS:

Lipopolysaccharide

UPR:

Unfolded protein response

Grp78:

Glucose-regulated protein 78

Grp94:

Glucose-regulated protein 94

HPRT:

Hypoxanthine-guanine phosphoribosyltransferase

IP3R: inositol 1:

4, 5-trisphosphate receptors

PARP:

Poly ADP ribose polymerase

NF-κB:

Nuclear factor-κB

MAPK:

Mitogen-activated protein kinase

IkB-α:

IkappaB-alpha

TNF-α:

Tumor necrosis factor-α

IL-1β:

Interleukin1β

IL-6:

Interleukin6

4-PBA:

4-phenylbutyrate

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

MPO:

Myeloperoxidase

ELISA:

Enzyme-linked immunosorbent assay

TLR4:

Toll-like receptor 4

BMM:

Bone marrow-derived macrophage

PAGE:

Polyacrylamide gel electrophoresis

TM:

Tunicamycin

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Acknowledgments

This work was supported by the China National Key Project of the Twelfth Five-year Plan (2012ZX10002004-006, 2012ZX10004904-003-001, 2013ZX10002002-006-001, 2012ZX10002005-003-003), the National Natural Science Foundation of China (81270532, 81372094,81300349), the Beijing Excellent Talents Training Funding (2011D003034000022), the Technology Foundation for Selected Overseas Chinese Scholar, the Ministry of Personnel of Beijing (2012), the Applied Research for the Clinical Characteristics of Capital (Z1211070010112167), and the Cooperation Research Project of CMU and Clinical (13JL33).

Conflict of Interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Beijing Artificial Liver Treatment & Training Center, Beijing YouAn Hospital, Capital Medical University, Beijing, China

    Feng Ren, Li Zhou, Xiangying Zhang, Tao Wen, Hongbo Shi, Bangxiang Xie, Zhuo Li, Dexi Chen & Zhongping Duan

  2. Qingdao Infectious Disease Hospital, Qingdao, Shandong Province, China

    Zheling Wang

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  1. Feng Ren
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  2. Li Zhou
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Correspondence to Zheling Wang or Zhongping Duan.

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Feng Ren and Li Zhou have contributed equally to this study.

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Ren, F., Zhou, L., Zhang, X. et al. Endoplasmic Reticulum Stress-Activated Glycogen Synthase Kinase 3β Aggravates Liver Inflammation and Hepatotoxicity in Mice with Acute Liver Failure. Inflammation 38, 1151–1165 (2015). https://doi.org/10.1007/s10753-014-0080-2

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