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Endoplasmic reticulum stress-induced hepatic stellate cell apoptosis through calcium-mediated JNK/P38 MAPK and Calpain/Caspase-12 pathways

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Abstract

Recent reports considered that it was the disturbance of calcium homeostasis and the accumulation of misfolded proteins in the endoplasmic reticulum (ER) that activated hepatic stellate cells (HSCs) apoptosis and promoted fibrosis resolution. However, the signal-transducing events that are activated by ER stress after HSCs activation were incompletely understood. In this study, we induced ER stress with thapsigargin (TG), and determined the activation of calpain and the cleavage of caspase by analyzing the protein levels and the correspondingly increased intracellular calcium levels and the induction of the proapoptotic transcription factor CHOP. Moreover, the phosphorylation of JNK and p38 MAPK were followed by the activation of the executioner caspases, caspase-3. As expected, preventing an increase in intracellular calcium levels using intracellular calcium chelators, EGTA, and BAPTA/AM, could substantially inhibit the phosphorylation of JNK and p38 MAPK, abolish the activation of calpains, namely caspase-12, caspase-9, and caspase-3, and provide significant protection for TG-treated activated HSCs. Interestingly, pretreatment with p38 MAPK inhibitor SB202190, JNK inhibitor SP600125, the pan-caspase inhibitor z-VAD-FMK, or calpain inhibitors calpeptin, significantly reduced the cell apoptosis and the cleavage of caspase-12 and caspase-3. However, pretreatment with z-VAD-FMK failed to reduce the activation of calpain. Additionally, pretreatment with SB202190 and SP600125 also decreased the expression of CHOP. Importantly, PDGF-induced collagen Col1α1 and α-smooth muscle actin (α-SMA), markers for the perpetuation phase of HSCs activation, were inhibited in TG-treated activated HSCs. These findings showed that the Calpain/Caspase-12 activation induced by ER stress and the JNK/p38 MAPK phosphorylation induced by the increase of intracellular calcium concentration releasing from ER are the novel signaling pathway underlying the molecular mechanism of fibrosis recovery.

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Abbreviations

α-SMA:

Alpha-smooth muscle actin

CHOP:

C/EBP homologues protein

ECM:

Extracellular matrix

ER:

Endoplasmic reticulum

ERK:

Extracellular signal-regulated protein kinase

HF:

Hepatic fibrosis

HSCs:

Hepatic stellate cells

JNK:

C-jun N-terminal kinase

MAPK:

Mitogen-activated protein kinase

siRNA:

Small interfering RNA

TG:

Thapsigargin

UPR:

Unfolded protein response

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Acknowledgments

This study is supported by the Chinese National Natural Science Foundation Project (No. 81102493, 81273526, 81072686) and the Specialized Research Fund for the Doctoral Program of Higher Education of China (No. 20103420120001).

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The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. School of Pharmacy, Institute for Liver Diseases, Anhui key laboratory of bioactivity of natural products, Anhui Medical University, Hefei, China

    Yan Huang, Xiaohui Li, Yarui Wang, Huan Wang, Cheng Huang & Jun Li

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  1. Yan Huang
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  2. Xiaohui Li
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  3. Yarui Wang
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  4. Huan Wang
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Correspondence to Jun Li.

Additional information

Y Huang and Xiaohui Li are co-first authors; they contributed equally to the work.

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Huang, Y., Li, X., Wang, Y. et al. Endoplasmic reticulum stress-induced hepatic stellate cell apoptosis through calcium-mediated JNK/P38 MAPK and Calpain/Caspase-12 pathways. Mol Cell Biochem 394, 1–12 (2014). https://doi.org/10.1007/s11010-014-2073-8

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