Effects of 2,2'-Azobis (2-Amidinopropane) Dihydrochloride-Induced Oxidative Stress on Cell Viability and Mitochondrial Function of Porcine Intestinal Epithelial Cells

LU Huijie; ZHANG Ying; TIAN Zhimei; RONG Ting; LIU Zhichang; DENG Dun; MA Xianyong

doi:10.3969/j.issn.1006-267x.2021.07.045

2021 , Vol. 33 >Issue 7: 4038 - 4048

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.07.045

MOLECULAR AND CELLULAR NUTRITION

Effects of 2,2'-Azobis (2-Amidinopropane) Dihydrochloride-Induced Oxidative Stress on Cell Viability and Mitochondrial Function of Porcine Intestinal Epithelial Cells

LU Huijie ,
ZHANG Ying ,
TIAN Zhimei ,
RONG Ting ,
LIU Zhichang ,
DENG Dun ,
MA Xianyong

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1. State Key Laboratory of Livestock and Poultry Breeding, Key Laboratory of Animal Nutrition and Feed Science in South China, Ministry of Agriculture and Rural Affairs, Guangdong Key Laboratory of Animal Breeding and Nutrition, Guangdong Engineering Technology Research Center of Animal Meat Quality and Safety Control and Evaluation, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
2. Maoming Branch,Guangdong Laboratory for Lingnan Modern Agriculture, Maoming 525000, China

Received date: 2020-12-02

Online published: 2021-07-06

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Abstract

The aim of this study was to evaluate the effects of 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress on cell viability and mitochondrial function of porcine intestinal epithelial cells (IPEC-J2 cells), and to establish a stable in vitro oxidative damage model. IPEC-J2 cells were treated with different concentrations (0, 30, 32 and 34 mmol/L) of AAPH for 24 h, the cell viability, cell proliferation, intracellular total reactive oxygen species (ROS) content, mitochondria membrane potential, mitochondrial DNA (mtDNA) copy number and the expression of mitochondrial function-related genes[mitochondrial transcription factor A (TFAM), mitochondrial transcription factor B1 (TFB1M), mitochondrial transcription factor B2 (TFB2M), uncoupling protein 1 (UCP1), uncoupling protein 2 (UCP2) and uncoupling protein 3 (UCP3)] were measured. The results showed that AAPH had a dosage and time dependent relationship on the viability of IPEC-J2 cells. Compared with the 0 mmol/L AAPH treatment, different concentrations (30, 32 and 34 mmol/L) of AAPH treatment significantly inhibited the proliferation of IPEC-J2 cells (P<0.05), the cell viability significantly decreased (P<0.05), the intracellular total ROS content significantly increased (P<0.05), the mRNA expression of mitochondrial function-related genes (TFAM, TFB1M, TFB2M, UCP1 and UCP3) significantly down-regulated (P<0.05), and the copy number of mtDNA significantly decreased (P<0.05). In summary, AAPH-induced oxidative damage leads to cell mitochondria dysfunction, resulting in decrease of cell viability and inhibited cell proliferation of IPEC-J2 cells.

Key words： AAPH; oxidative stress; intestinal epithelial cells; mitochondria

Cite this article

LU Huijie , ZHANG Ying , TIAN Zhimei , RONG Ting , LIU Zhichang , DENG Dun , MA Xianyong . Effects of 2,2'-Azobis (2-Amidinopropane) Dihydrochloride-Induced Oxidative Stress on Cell Viability and Mitochondrial Function of Porcine Intestinal Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2021 , 33(7) : 4038 -4048 . DOI: 10.3969/j.issn.1006-267x.2021.07.045

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