Effects of Lactobacillus acidophilus on Antioxidative Ability and Expression of Tight Junction Protein in Intestinal Epithelial Cells of Piglets under Oxidative Stress

LUO Bowen; ZOU Tiande; CHEN Lilin; ZENG Yongdi; GUO Xiaobo; YOU Jinming

doi:10.3969/j.issn.1006-267x.2020.05.019

2020 , Vol. 32 >Issue 5: 2108 - 2115

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.05.019

SWINE NUTRITION AND FEED

Effects of Lactobacillus acidophilus on Antioxidative Ability and Expression of Tight Junction Protein in Intestinal Epithelial Cells of Piglets under Oxidative Stress

LUO Bowen ,
ZOU Tiande ,
CHEN Lilin ,
ZENG Yongdi ,
GUO Xiaobo ,
YOU Jinming

Expand

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

Received date: 2019-11-14

Online published: 2020-05-15

Fold

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.

Key words： Lactobacillus acidophilus; IPEC-J2; antioxidant; tight junction protein

Cite this article

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[J]. Chinese Journal of Animal Nutrition, 2020 , 32(5) : 2108 -2115 . DOI: 10.3969/j.issn.1006-267x.2020.05.019

References

[1] WANG N,TANG X C,MA W P,et al.Tu1852 resveratrol ameliorates oxidative stress-induced intestinal epithelial barrier dysfunction by upregulating heme oxygenase-1 expression[J].Gastroenterology,2015,148(4):2522-2534.
[2] 赵玉蓉,王耀东.牛膝多糖抑制沙门氏菌侵染猪小肠上皮细胞的研究[J].动物营养学报,2018,30(12):5083-5088.
[3] 陈建永,徐虹,施琳琳,等.正肝汤对肝硬化大鼠肠道紧密连接蛋白OCLN和ZO-1表达的影响[J].中华中医药学刊,2015,33(12):2991-2994.
[4] GONZÁLEZ-MARISCAL L,LECHUGA S,GARAY E.Role of tight junctions in cell proliferation and cancer[J].Progress in Histochemistry and Cytochemistry,2008,42(1):1-57.
[5] KIM S,KIM G H.Roles of claudin-2,ZO-1 and occludin in leaky HK-2 cells[J].PLoS One,2017,12(12):e0189221.
[6] 邢爽,冯京海.乳酸杆菌对肠道上皮紧密连接蛋白的调控[J].动物营养学报,2019,31(4):1540-1546.
[7] 崔志文.鼠李糖乳杆菌影响仔猪肠道屏障功能的研究[D].博士学位论文.杭州:浙江大学,2013.
[8] AHRNE S,HAGSLATT M L J.Effect of lactobacilli on paracellular permeability in the gut[J].Nutrients,2011,3(1):104-117.

[9] DAI C,ZHAO D H,JIANG M.VSL#3 probiotics regulate the intestinal epithelial barrier in vivo and in vitro via the p38 and ERK signaling pathways[J].International Journal of Molecular Medicine,2012,29(2):202-208.
[10] 雷一慧.乳凝集素改善坏死性小肠结肠炎肠道通透性及损伤修复的研究[D].硕士学位论文.上海:上海交通大学,2015.
[11] 王鑫,张萌萌,姜宁,等.宿主防御肽调节动物肠道屏障功能的研究进展[J].动物营养学报,2018,30(9):3410-3416.
[12] HANDAYANINGSIH A E,IGUCHI G,FUKUOKA H,et al.Reactive oxygen species play an essential role in IGF-Ⅰ signaling and IGF-Ⅰ-induced myocyte hypertrophy in C2C12 myocytes[J].Endocrinology,2011,152(3):912-921.

[13] ZHU L H,ZHAO K L,CHEN X L,et al.Impact of weaning and an antioxidant blend on intestinal barrier function and antioxidant status in pigs[J].Journal of Animal Science,2012,90(8):2581-2589.

[14] LYKKESFELDT J,SVENDSEN O.Oxidants and antioxidants in disease:oxidative stress in farm animals[J].The Veterinary Journal,2014,173(3):502-511.
[15] KIM Y J,KIM E H,HAHM K B.Oxidative stress in inflammation-based gastrointestinal tract diseases:challenges and opportunities[J].Journal of Gastroenterology and Hepatology,2012,27(6):1004-1010.

[16] RYTER S WKIM H P,HOETZEL A,et al.Mechanisms of cell death in oxidative stress[J].Antioxidants & Redox Signaling,2007,9(1):49-89.

[17] PASZTI-GERE E,CSIBRIK-NEMETH E,SZEKER K,et al.Acute oxidative stress affects IL-8 and TNF-α expression in IPEC-J2 porcine epithelial cells[J].Inflammation,2012,35(3):994-1004.

[18] YANG Y,KARAKHANOVA S,WERNER J,et al.Reactive oxygen species in cancer biology and anticancer therapy[J].Current Medicinal Chemistry,2013,20(30):3677-3692.

[19] BRUNO-BÁRCENA J M,ANDRUS J M,LIBBY S L,et al.Expression of a heterologous manganese superoxide dismutase gene in intestinal lactobacilli provides protection against hydrogen peroxide toxicity[J].Applied & Environmental Microbiology,2004,70(8):4702-4710.

[20] SERRANO L M,MOLENAAR D,WELS M,et al.Thioredoxin reductase is a key factor in the oxidative stress response of Lactobacillus plantarum WCFS1[J].Microbial Cell Factories,2007,6(1):29.
[21] ALFONSO-PRIETO M,BIARNÉS X,VIDOSSICH P,et al.The molecular mechanism of the catalase reaction[J].Journal of the American Chemical Society,2009,131(33):11751-11761.

[22] 白明,孟祥晨.益生菌抗氧化活性及菌体抗氧化相关成分的分析[J].食品与发酵工业,2009,35(5):6-11.
[23] HARHAJ N S,ANTONETTI D A.Regulation of tight junctions and loss of barrier function in pathophysiology[J].International Journal of Biochemistry & Cell Biology,2004,36(7):1206-1237.

[24] 张建龙,丘富安,董星,等.猴头菇多糖对氧化应激状态下仔猪回肠形态结构及通透性的影响[J].畜牧兽医学报,2018,49(1):203-210.
[25] UKENA S N,SINGH A,DRINGENBERG U,et al.Probiotic Escherichia coli nissle 1917 inhibits leaky gut by enhancing mucosal integrity[J].PLoS One,2007,2(12):e1308.
[26] PATEL R M,MYERS L S,KURUNDKAR A R,et al.Probiotic bacteria induce maturation of intestinal Claudin 3 expression and barrier function[J].The American Journal of Pathology,2012,180(2):626-635.

[27] ANDERSON R C,COOKSON A L,MCNABB W C,et al.Lactobacillus plantarum MB452 enhances the function of the intestinal barrier by increasing the expression levels of genes involved in tight junction formation[J].BMC Microbiology,2010,10:316.
[28] 张俊,朱建津,刘江英,等.益生菌发酵产物对肉鸡肠道形态和肠道屏障功能的影响[J].浙江农业学报,2016,28(10):1657-1662.
[29] 刘理,杨宗城,黎鳌.紧密连接的分子基础及其调控[J].国外医学(生理、病理科学与临床分册),1999,19(1):75-77.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References