Effects of Lactobacillus acidophilus on Antioxidative Ability and Expression of Tight Junction Protein in Intestinal Epithelial Cells of Piglets under Oxidative Stress
LUO Bowen1, ZOU Tiande1, CHEN Lilin1,2, ZENG Yongdi1, GUO Xiaobo1, YOU Jinming1
1. Jiangxi Province Key Laboratory of Animal Nutrition, Engineering Research Center of Feed Development, Jiangxi Agricultural University, Nanchang 330045, China;
2. Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China
Abstract：The purpose of this study was to investigate the effects of Lactobacillus acidophilus on antioxidative ability and expression of tight junction protein (occludin, ZO-1 and claudin-1) in intestinal epithelial cells of piglets (IPEC-J2) under oxidative stress. Firstly, the IPEC-J2 were treated with different H2O2 concentrations, including 0 (control), 0.6, 0.8, 1.0, 1.2, 1.4, 1.6, 1.8 and 2.0 mmol/L, with 6 replications in each group. The optimum concentration of H2O2 was determined according to cell activity. And then, the 1.2 mmol/L H2O2 were used to induce oxidative stress. The cells were divided into 7 groups, and added 0(control), 1.0×105, 1.0×106, 1.0×107, 1.0×108, 1.0×109 and 1.0×1010 CFU/mL Lactobacillus acidophilus, respectively, with 6 replications in each group, and treated for 24 h, analyzing the antioxidant ability of Lactobacillus acidophilus in IPEC-J2 under oxidative stress. At last, 1.2 mmol/L H2O2 was selected to establish the IPEC-J2 oxidative stress model, and then 1.0×109 CFU/mL Lactobacillus acidophilus was added to culture cells for 24 h, and then cells was divided into four groups:the control group (adding equivalent phosphate buffer), the H2O2 oxidative stress group, the Lactobacillus acidophilus group, and the Lactobacillus acidophilus+H2O2 group (adding H2O2 first and then Lactobacillus acidophilus), with 6 replications in each group. The expressions of epithelial tight junction proteins (occludin, ZO-1 and claudin-1) were measured. The results showed as follows:1) compared with the control group, at the concentration of H2O2 ≥ 0.6 mmom/L, the cell activity was significantly reduced (P<0.05). Compared with control group and 0.6 to 1.0 mmol/L H2O2 groups, the activities of cells were decreased significantly at the concentration between 1.2 and 1.8 mmol/L H2O2 (P<0.05). 2) Compared with control group, at the concentration of Lactobacillus acidophilus ≥ 1.0×108 CFU/mL, the cell activity was significantly increased (P<0.05). 3) Under oxidative stress, with the increase of Lactobacillus acidophilus concentration, cell activity was increaseed gradually. Compared with control group, at the concentration of Lactobacillus acidophilus ≤ 1.0×107 CFU/mL, the cell activity was significantly decreased (P<0.05), at the concentration of Lactobacillus acidophilus ≥ 1.0×109 CFU/mL, cell activity was increased significantly (P<0.05). 4) Compared with control group, at the concentration of Lactobacillus acidophilus ≥ 1.0×106 CFU/mL, the content of malonaldehyde (MDA) in cells was significantly reduced (P<0.05). Compared with control group, at the concentration of Lactobacillus acidophilus ≥ 1.0×107 CFU/mL, superoxide dismutase (SOD) activity was significantly increased (P<0.05), and glutathion peroxidase (GSH-Px) activity in cells in other groups was significantly increased compared with control group (P<0.05). 5) Compared with the H2O2 oxidative stress group, the protein expressions of ZO-1, occludin and claudin proteins in Lactobacillus acidophilus group and Lactobacillus acidophilus+H2O2 group were significantly decreased (P<0.05). Compared with the control group and H2O2 oxidative stress group, claudin protein expression in Lactobacillus acidophilus group and Lactobacillus acidophilus+H2O2 group was significantly decreased (P<0.05). In conclusion, Lactobacillus acidophilus can promote the growth of cells, enhance the antioxidant ability of cells, and reduce the expression of intracytoplasmic tight junction protein.
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LUO Bowen, ZOU Tiande, CHEN Lilin, ZENG Yongdi, GUO Xiaobo, YOU Jinming. Effects of Lactobacillus acidophilus on Antioxidative Ability and Expression of Tight Junction Protein in Intestinal Epithelial Cells of Piglets under Oxidative Stress. Chinese Journal of Animal Nutrition, 2020, 32(5): 2108-2115.
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