[1]李若楠,康瑞芬,沈丹,等.谷氨酰胺对呕吐毒素诱导IPEC-J2细胞凋亡和炎症的影响[J].南京农业大学学报,2020,43(4):740-747.[doi:10.7685/jnau.201907009]
 LI Ruonan,KANG Ruifen,SHEN Dan,et al.Effects of glutamine on deoxynivalenol induced apoptosis and inflammation of IPEC-J2 cells[J].Journal of Nanjing Agricultural University,2020,43(4):740-747.[doi:10.7685/jnau.201907009]
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谷氨酰胺对呕吐毒素诱导IPEC-J2细胞凋亡和炎症的影响()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
43卷
期数:
2020年4期
页码:
740-747
栏目:
动物科学
出版日期:
2020-07-13

文章信息/Info

Title:
Effects of glutamine on deoxynivalenol induced apoptosis and inflammation of IPEC-J2 cells
作者:
李若楠 康瑞芬 沈丹 戴鹏远 唐倩 李春梅
南京农业大学动物科技学院, 江苏 南京 210095
Author(s):
LI Ruonan KANG Ruifen SHEN Dan DAI Pengyuan TANG Qian LI Chunmei
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
关键词:
谷氨酰胺呕吐毒素猪空肠上皮细胞炎症凋亡
Keywords:
glutaminedeoxynivalenolintestinal porcine epithelial cell(IPEC-J2 cell)inflammationapoptosis
分类号:
S828.9
DOI:
10.7685/jnau.201907009
摘要:
[目的] 本试验以猪空肠上皮细胞(IPEC-J2)为模型,探讨谷氨酰胺(Gln)对呕吐毒素(DON)诱导的IPEC-J2细胞凋亡和炎症的影响。[方法] 通过MTT方法测定细胞活力来选择适宜的Gln浓度,以不添加Gln和DON的细胞为空白对照组,试验组分别为0.75 mmol·L-1 Gln组,2.0 μg·mL-1 DON组和2.0 μg·mL-1 DON+0.75 mmol·L-1 Gln组,处理24 h后测定各组细胞凋亡率、活性氧自由基(ROS)及细胞凋亡和炎症相关基因和蛋白的表达水平。[结果] 与对照组相比,DON处理24 h细胞凋亡比例和ROS含量(ROS荧光密度/细胞总数)显著升高(P<0.01);与DON组相比,DON+Gln组显著降低细胞的凋亡比例和ROS含量(P<0.01)。与对照组相比,DON组上调IPEC-J2细胞白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、环氧合酶2(COX-2)及肿瘤坏死因子α(TNF-α)炎症相关基因和Caspase-3Caspase-8凋亡相关基因的表达水平;与DON组相比,DON+Gln组下调IPEC-J2细胞IL-1β、COX-2、Caspase-3、Caspase-8、BAKBcl-2基因的表达水平。蛋白免疫荧光结果显示,与对照组相比,DON组上调IPEC-J2细胞Caspase-3,核转录因子κB(NF-κB)和磷酸化核转录因子κB(phospho-NF-κB)蛋白的表达,而添加Gln后(DON+Gln组)下调了相关蛋白的表达。[结论] Gln通过清除DON诱导的IPEC-J2细胞中过量的ROS及调节炎症和凋亡相关基因及蛋白的表达量来缓解由DON引起的肠道上皮细胞损伤。
Abstract:
[Objectives] In this study,intestinal porcine epithelial cells(IPEC-J2 cells) were used as a model to investigate the effects of glutamine(Gln) on deoxynivalenol(DON) induced apoptosis and inflammation of IPEC-J2 cells.[Methods] The appropriate Gln concentration was selected by MTT cell viability assay,and the cells without Gln and DON were used as the control group. The treated groups were 0.75 mmol·L-1 Gln,2.0 μg·mL-1 DON and 2.0 μg·mL-1 DON+0.75 mmol·L-1 Gln,respectively. After 24 h,the apoptosis,reactive oxygen species(ROS),some genes and proteins associated with apoptosis and inflammation were measured.[Results] Compared with the control group,the percentage of apoptosis and ROS content(ROS fluorescence density/cell total) significantly increased after DON treatment for 24 h(P<0.01). Compared with the DON group,DON+Gln significantly decreased the percentage of apoptosis and ROS content(P<0.01). Compared with the control group,DON up-regulated the expression of genes involved in inflammation and apoptosis,such as interleukin-1β(IL-1β),interleukin-6(IL-6),cyclooxygenase 2(COX-2),tumor necrosis factor alpha(TNF-α),Caspase-3,Caspase-8;DON+Gln down-regulated the expression of IL-1β,COX-2,Caspase-3,Caspase-8,BAK and Bcl-2 genes compared with DON group. Protein immunofluorescence revealed that compared with the control group,DON up-regulated the expression of Caspase-3,nuclear factor kappa B(NF-κB) and phosphorylated nuclear factor kappa B(phospho-NF-κB) protein,and Gln down-regulate the expression of related proteins.[Conclusions] Gln alleviated the damage of IPEC-J2 cells caused by DON by decreasing ROS production and regulating the expression of genes and protein involved in inflammation and apoptosis.

参考文献/References:

[1] Li R,Li Y,Su Y,et al. Short-term ingestion of deoxynivalenol in naturally contaminated feed alters piglet performance and gut hormone secretion[J]. Animal Science Journal,2018,89(8):1134-1143.
[2] Ji F,Xu J,Liu X,et al. Natural occurrence of deoxynivalenol and zearalenone in wheat from Jiangsu Province,China[J]. Food Chemistry,2014,157:393-397.
[3] 龚阿琼,全明旭,周锋,等. 2017年上半年我国饲料原料及配合饲料霉菌毒素检测分析[J]. 中国饲料,2017(20):41-43. Gong A Q,Quan M X,Zhou F,et al. Detection and analysis of mycotoxin in feed ingredients and compound feed in China in the first half of 2017[J]. Chinese Feed,2017(20):41-43(in Chinese with English abstract).
[4] Wu Q,Ku A?1 a K,Humpf H U,et al. Fate of deoxynivalenol and deoxynivalenol-3-glucoside during cereal-based thermal food processing:a review study[J]. Mycotoxin Research,2017,33(1):79-91.
[5] González-Castro A M,Martínez C,Salvo-Romero E,et al. Mucosal pathobiology and molecular signature of epithelial barrier dysfunction in the small intestine in irritable bowel syndrome[J]. Journal of Gastroenterology and Hepatology,2017,32(1):53-63.
[6] Awad W A,Zentek J. The feed contaminant deoxynivalenol affects the intestinal barrier permeability through inhibition of protein synthesis[J]. Archives of Toxicology,2015,89(6):961-965.
[7] Bracarense A P,Lucioli J,Grenier B,et al. Chronic ingestion of deoxynivalenol and fumonisin,alone or in interaction,induces morphological and immunological changes in the intestine of piglets[J]. British Journal of Nutrition,2012,107(12):1776-1786.
[8] Kang R,Li R,Dai P,et al. Deoxynivalenol induced apoptosis and inflammation of IPEC-J2 cells by promoting ROS production[J]. Environmental Pollution,2019,251:689-698.
[9] Liu Y,Xiong X,Tan B E,et al. Nutritional intervention for the intestinal development and health of weaned pigs[J]. Frontiers in Veterinary Science,2019,6:46-59.
[10] Wang H,Zhang C,Wu G Y,et al. Glutamine enhances tight junction protein expression and modulates corticotropin-releasing factor signaling in the jejunum of weanling piglets[J]. Journal of Nutrition,2015,145(1):25-31.
[11] Xing S,Zhang B L,Lin M,et al. Effects of alanyl-glutamine supplementation on the small intestinal mucosa barrier in weaned piglets challenged with lipopolysaccharide[J]. Canadian Journal of Animal Science,2017,98(1):144-155.
[12] Wang B,Wu G,Zhou Z,et al. Glutamine and intestinal barrier function[J]. Amino Acids,2015,47(10):2143-2154.
[13] Ding Z,Li W,Huang J,et al. Dietary alanyl-glutamine and vitamin E supplements could considerably promote the expression of GPx and PPARα genes,antioxidation,feed utilization,growth,and improve composition of juvenile cobia[J]. Aquaculture,2017,470:95-102.
[14] Evans M E,Jones D P,Ziegler T R. Glutamine prevents cytokine-induced apoptosis in human colonic epithelial cells[J]. Journal of Nutrition,2003,133(10):3065-3071.
[15] Kim M H,Kim H. The roles of glutamine in the intestine and its implication in intestinal diseases[J]. International Journal of Molecular Sciences,2017,18(5):1051-1066.
[16] Chen G,Shi J,Qi M,et al. Glutamine decreases intestinal nuclear factor kappa B activity and pro-inflammatory cytokine expression after traumatic brain injury in rats[J]. Inflammation Research,2008,57(2):57-64.
[17] 黄杰,刘京,马娜娜,等. 谷氨酰胺对急性肺损伤小鼠MUC5AC的调节及其对肺脏的保护作用[J]. 畜牧与兽医,2019,51(4):97-103. Huang J,Liu J,Ma N N,et al. Regulative and protective effects of glutamine on the expression of MUC5AC[J]. Animal Husbandry & Veterinary Medicine,2019,51(4):97-103(in Chinese with English abstract).
[18] Zhou J,Fan S,Cao Y,et al. Tumor necrosis factor-α suppresses the protein fractional synthesis rate of the small intestine stimulated by glutamine in rats[J]. Experimental and Therapeutic Medicine,2015,9(2):547-552.
[19] Sverrisson K,Axelsson J,Rippe A,et al. Acute reactive oxygen species(ROS)-dependent effects of IL-1β,TNF-α,and IL-6 on the glomerular filtration barrier(GFB)in vivo[J]. American Journal of Physiology Renal Physiology,2015,309(9):F800-F806.

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备注/Memo

备注/Memo:
收稿日期:2019-07-06。
基金项目:国家自然科学基金项目(31772648)
作者简介:李若楠,硕士研究生。
通信作者:李春梅,教授,博士,主要从事饲料卫生、畜禽环境生理与营养调控研究,E-mail:chunmeili@njau.edu.cn。
更新日期/Last Update: 1900-01-01