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Protective Effect of Selenoprotein X Against Oxidative Stress-Induced Cell Apoptosis in Human Hepatocyte (LO2) Cells via the p38 Pathway

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Abstract

Oxidative stress, as mediated by ROS (reactive oxygen species), is a significant factor in initiating the cells damaged by affecting cellular macromolecules and impairing their biological functions; SelX, a selenoprotein also known as MsrB1 belonging to the methionine sulfoxide reductase (Msr) family, is the redox repairing enzyme and involved in redox-related functions. In order to more precisely analyze the relationship between oxidative stress, cell oxidative damage, and SelX, we stably overexpressed porcine Selx full-length cDNA in human normal hepatocyte (LO2) cells. Cell viability, cell apoptosis rate, intracellular ROS, and the expression levels of mRNA or protein of apoptosis-related genes under H2O2-induced oxidative stress were detected. We found that overexpression of SelX can prevent the oxidative damage caused by H2O2 and propose that the main mechanism underlying the protective effects of SelX is the inhibition of LO2 cell apoptosis. The results revealed that overexpressed SelX reduced the H2O2-induced intracellular ROS generation, inhibited the H2O2-induced upregulation of Bax and downregulation of Bcl-2, and increased the mRNA and protein ratio of Bcl-2/Bax. Furthermore, it inhibited H2O2-induced p38 MAPK phosphorylation. Taken together, our findings suggested that SelX played important roles in protecting LO2 cells against oxidative damage and that its protective effect is partly via the p38 pathway by acting as a ROS scavenger.

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Abbreviations

Msr:

Methionine sulfoxide reductase

SelX:

Selenoprotein X

ROS:

Reactive oxygen species

LO2:

Human normal hepatocyte;

p38 MAPK:

p38 mitogen-activated protein kinase

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethyl sulfoxide

ECL:

Electrochemiluminescence

PVDF:

Polyvinylidene fluoride

SDS:

Sodium dodecyl sulfate

SE:

Standard error

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Acknowledgements

This work was supported partly by the National Natural Science Foundation of China (No 31272468 and 31072043) and by a Research Funding provided by Sichuan Longda animal husbandry science and technology co., Ltd. (No: 2015SCLD001).

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Authors and Affiliations

  1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China

    Jia-Yong Tang, Ai-Hua He, Gang Jia, Guang-Mang Liu, Xiao-Ling Chen, Jing-Yi Cai, Hai-Ying Shang & Hua Zhao

  2. Trace Element Research Center, Sichuan Agricultural University, No 211 Huimin Road, Chengdu, Sichuan, 611130, China

    Jia-Yong Tang & Hua Zhao

  3. College of Life Science, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China

    Jin-Qiu Liao

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  1. Jia-Yong Tang
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Correspondence to Hua Zhao.

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All authors read and approved the final manuscript. The authors have no conflicts of interest to disclose.

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Ai-Hua He is the co-first author.

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Tang, JY., He, AH., Jia, G. et al. Protective Effect of Selenoprotein X Against Oxidative Stress-Induced Cell Apoptosis in Human Hepatocyte (LO2) Cells via the p38 Pathway. Biol Trace Elem Res 181, 44–53 (2018). https://doi.org/10.1007/s12011-017-1025-z

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