Effects of Glutamine on Oxidative Damage and Apoptosis of Human Enterocyte-Like HT-29 Cells Induced by Hydrogen Peroxide

XIAO Yingping, GUI Guohong, CHEN Anguo, HE Xiangxiang, LI Kaifeng, YANG Hua

doi:10.3969/j.issn.1006-267x.2017.08.016

2017 , Vol. 29 >Issue 8: 2734 - 2742

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.08.016

Molecular Nutrition

Effects of Glutamine on Oxidative Damage and Apoptosis of Human Enterocyte-Like HT-29 Cells Induced by Hydrogen Peroxide

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1. Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
2. State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
3. College of Animal Science, Guizhou University, Guiyang 550025, China;
4. College of Animal Sciences, Zhejiang University, Hangzhou 310058, China

Received date: 2017-02-01

Online published: 2017-08-02

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Abstract

This experiment was conducted to study effects of glutamine (Gln) on oxidative damage and apoptosis of human enterocyte-like HT-29 cells induced by hydrogen peroxide (H₂O₂), and to explore the antioxidant function of Gln and its mechanisms. HT-29 cells I were induced by different concentrations [0 (control group), 0.5, 2.0 and 10.0 mmol/L] Gln and 0.35 mmol/L H₂O₂ for 12, 24 and 32 h, the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells were measured, and the mRNA relative expression of related to cell apoptosis gene were also assessed by RT-qPCR. Furthermore, the HT-29 cells were dyed with the Annexin V-FITC/PI double staining method, and the apoptosis of cells was determined by the flow cytometer. The results showed as follows: 1) after treated 24 h, the SOD activity in 0.5 and 2.0 mmol/L Gln groups was significantly higher than that in control group (P<0.05); after treated 32 h, the SOD activity in 0.5 and 2.0 mmol/L Gln groups was significantly higher than that in control group (P<0.05), and the MDA content was significantly lower than that in control group (P<0.05). 2) After treated 12 h, there were no significant differences on the mRNA relative expressions of cysteinyl aspartate specific proteinase-3 (Caspase-3) and B cell lymphoma/leukemia-2 associated X protein (Bax) among all groups (P>0.05). Compared with the control group, the mRNA relative expression of nuclear factor kappa B (NF-κB) in 0.5 mmol/L Gln group was significantly decreased (P<0.05), the mRNA relative expression of B cell lymphoma/leukemia-2 (Bcl-2) in 0.5 and 2.0 mmol/L Gln groups was significantly increased (P<0.05). After treated 24 h, compared with the control group, the mRNA relative expressions of Caspase-3, NF-κB and Bax in 0.5, 2.0 and 10.0 mmol/L Gln groups were significantly decreased (P<0.05), while the mRNA relative expression of Bcl-2 was significantly increased (P<0.05). After treated 32 h, compared with the control group, the mRNA relative expressions of FAS, Caspase-3, NF-κB and Bax in 2.0 mmol/L Gln group were significantly decreased (P<0.05), while the mRNA relative expression of Bcl-2 was significantly increased (P<0.05); the mRNA relative expressions of FAS, Caspase-3, NF-κB and Bax in 10.0 mmol/L Gln group were significantly decreased (P<0.05). 3) After treated 24 h, compared with the control group, the living cells number in Gln groups increased by 5.32% to 11.97%, and the necrotic cells number decreased by 6.75% to 12.66%. After treated 32 h, compared with the control group, the living cells number in Gln groups increased by 1.39% to 7.63%, and the necrotic cells number decreased by 3.40% to 4.57%. It is concluded that glutamine can restrain the oxidant damage of HT-29 induced by H₂O₂ and decrease the cells apoptosis.

Key words： glutamine; oxidative damage; cell apoptosis; HT-29 cell; hydrogen peroxide

Cite this article

XIAO Yingping, GUI Guohong, CHEN Anguo, HE Xiangxiang, LI Kaifeng, YANG Hua . Effects of Glutamine on Oxidative Damage and Apoptosis of Human Enterocyte-Like HT-29 Cells Induced by Hydrogen Peroxide[J]. Chinese Journal of Animal Nutrition, 2017 , 29(8) : 2734 -2742 . DOI: 10.3969/j.issn.1006-267x.2017.08.016

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