[1]萧晟,刘丹丹,甘芳,等.脱氧雪腐镰刀菌烯醇对IPEC-J2细胞增殖、凋亡和屏障功能的影响及其自噬机制[J].南京农业大学学报,2019,42(4):746-751.[doi:10.7685/jnau.201901044]
 XIAO Sheng,LIU Dandan,GAN Fang,et al.Effects of deoxynivalenol on the proliferation,apoptosis and barrier function of IPEC-J2 cells and its autophagy mechanisms[J].Journal of Nanjing Agricultural University,2019,42(4):746-751.[doi:10.7685/jnau.201901044]
点击复制

脱氧雪腐镰刀菌烯醇对IPEC-J2细胞增殖、凋亡和屏障功能的影响及其自噬机制()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
42卷
期数:
2019年4期
页码:
746-751
栏目:
动物科学
出版日期:
2019-07-08

文章信息/Info

Title:
Effects of deoxynivalenol on the proliferation,apoptosis and barrier function of IPEC-J2 cells and its autophagy mechanisms
作者:
萧晟12 刘丹丹2 甘芳2 黄克和2
1. 芜湖职业技术学院, 安徽 芜湖 241003;
2. 南京农业大学动物医学院, 江苏 南京 210095
Author(s):
XIAO Sheng12 LIU Dandan2 GAN Fang2 HUANG Kehe2
1. Wuhu Institute of Technology, Wuhu 241003, China;
2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
关键词:
脱氧雪腐镰刀菌烯醇仔猪空肠上皮细胞细胞凋亡紧密连接细胞自噬
Keywords:
deoxynivalenolIPEC-J2 cellsapoptosistight junctionautophagy
分类号:
S856.4
DOI:
10.7685/jnau.201901044
摘要:
[目的]本试验旨在研究脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)对体外培养的仔猪空肠上皮细胞(IPEC-J2)增殖、凋亡和屏障功能的影响,探讨DON的肠道毒性作用及相关机制。[方法]以不同浓度(0、0.5、1、2、4、8、和16 μmol·L-1) DON处理IPEC-J2细胞24 h后,采用MTT、Hoechst 33258、Western blot、Millipore-ERS和IFA法检测DON对IPEC-J2细胞增殖、凋亡、屏障功能和自噬水平的影响;在4 μmol·L-1 DON处理组添加自噬激活剂雷帕霉素(Rapa)来提高细胞的自噬水平,用上述方法检测Rapa对DON引起的IPEC-J2细胞增殖毒性、凋亡和屏障功能的影响。[结果]与对照组相比,当DON浓度为4、8和16 μmol·L-1时,细胞活性显著(P<0.05)或极显著(P<0.01)下降。4和8 μmol·L-1 DON处理凋亡细胞数量和凋亡相关蛋白Cleaved-Caspase3表达显著增加(P<0.01),细胞跨膜电阻(TEER)值和紧密连接蛋白Occludin表达显著下降(P<0.05),自噬相关蛋白LC3-Ⅱ和Atg5表达显著下降(P<0.05)。添加Rapa可以显著缓解DON引起的细胞毒性、凋亡和屏障功能障碍。[结论]DON通过抑制自噬引起IPEC-J2细胞增殖下降、凋亡增加和屏障功能障碍。
Abstract:
[Objectives]The purpose of this study was to investigate the effects of deoxynivalenol(DON)on proliferation,apoptosis and barrier function of intestinal porcine epithelial cells(IPEC-J2)in vitro,and its autophagy mechanisms.[Methods]IPEC-J2 cells were treated with different concentrations of DON(0,0.5,1,2,4,8 and 16 μmol·L-1)for 24 h. Cell proliferation,apoptosis,barrier damage and autophgay were detected by thiazolyl blue tetrazolium bromide(MTT),Hoechst 33258,Western blot,Millipore-ERS and immunofluorescence(IFA). After autophagy activator rapamycin(Rapa)inducing an increased level of autophagy in 4 μmol·L-1 DON group,the effects of DON on cell proliferation,apoptosis and barrier damage of IPEC-J2 cells were assayed.[Results]The results showed that DON(4,8 and 16 μmol·L-1)significantly reduced the cell viability(P<0.05 or P<0.01). DON(4 and 8 μmol·L-1)increased the number of apoptotic cells and the expression of apoptosis-related protein Cleaved-Caspase3(P<0.01),and decreased cell transmembrane resistance(TEER)value and the expression of tight junction protein Occludin(P<0.05). DON(4 and 8 μmol·L-1)also significantly decreased the expressions of autophagy-related protein LC3-Ⅱ and Atg5(P<0.05). However,the Rapa could significantly alleviate the cell toxicity,apoptosis and barrier disorder induced by DON.[Conclusions]In conclusion,DON can decrease cell proliferation and barrier dysfunction,and increase apoptosis by inhibiting autophagy in IPEC-J2 cells.

参考文献/References:

[1] Ebrahem M,Kersten S,Valenta H,et al. Effects of feeding deoxynivalenol(DON)-contaminated wheat to laying hens and roosters of different genetic background on the reproductive performance and health of the newly hatched chicks[J]. Mycotoxin Res,2014,30(3):131-140.
[2] Wang S,Yang J,Zhang B,et al. Deoxynivalenol impairs porcine intestinal host defense peptide expression in weaned piglets and IPEC-J2 cells[J]. Toxins(Basel),2018,10(12):541-556.
[3] Pierron A,Alassane-Kpembi I,Oswald I P. Impact of two mycotoxins deoxynivalenol and fumonisin on pig intestinal health[J]. Porcine Health Manag,2016,2:21-28.
[4] Lucke A,Bohm J,Zebeli Q,et al. Dietary deoxynivalenol and oral lipopolysaccharide challenge differently affect intestinal innate immune response and barrier function in broiler chickens[J]. J Anim Sci,2018,96(12):5134-5143.
[5] Vancamelbeke M,Vermeire S. The intestinal barrier:a fundamental role in health and disease[J]. Expert Rev Gastroenterol Hepatol,2017,11(9):821-834.
[6] Luissint A C,Parkos C A,Nusrat A. Inflammation and the intestinal barrier:leukocyte-epithelial cell interactions,cell junction remodeling,and mucosal repair[J]. Gastroenterology,2016,151(4):616-632.
[7] Kim H J,Lee S,Jung J U. When autophagy meets viruses:a double-edged sword with functions in defense and offense[J]. Semin Immunopathol,2010,32(4):323-341.
[8] Sun Y,Yu S,Ding N,et al. Autophagy benefits the replication of Newcastle disease virus in chicken cells and tissues[J]. J Virol,2014,88(1):525-537.
[9] Levine B,Kroemer G. Biological functions of autophagy genes:a disease perspective[J]. Cell,2019,176(1/2):11-42.
[10] Chen X,Xu J,Liu D,et al. The aggravating effect of selenium deficiency on T-2 toxin-induced damage on primary cardiomyocyte results from a reduction of protective autophagy[J]. Chem Biol Interact,2019,300:27-34.
[11] Cadwell K. Crosstalk between autophagy and inflammatory signalling pathways:balancing defence and homeostasis[J]. Nat Rev Immunol,2016,16(11):661-675.
[12] Matsuzawa-Ishimoto Y,Shono Y,Gomez L E,et al. Autophagy protein ATG16L1 prevents necroptosis in the intestinal epithelium[J]. J Exp Med,2017,214(12):3687-3705.
[13] Ghareeb K,Awad W A,Bohm J,et al. Impacts of the feed contaminant deoxynivalenol on the intestine of monogastric animals:poultry and swine[J]. J Appl Toxicol,2015,35(4):327-337.
[14] Meurens F,Summerfield A,Nauwynck H,et al. The pig:a model for human infectious diseases[J]. Trends Microbiol,2012,20(1):50-57.
[15] Patterson J K,Lei X G,Miller D D. The pig as an experimental model for elucidating the mechanisms governing dietary influence on mineral absorption[J]. Exp Biol Med,2008,233(6):651-664.
[16] Adesso S,Autore G,Quaroni A,et al. The food contaminants nivalenol and deoxynivalenol induce inflammation in intestinal epithelial cells by regulating reactive oxygen species release[J]. Nutrients,2017,9(12):1343-1359.
[17] Pierron A,Mimoun S,Murate L S,et al. Intestinal toxicity of the masked mycotoxin deoxynivalenol-3-beta-D-glucoside[J]. Arch Toxicol,2016,90(8):2037-2046.
[18] Awad W A,Aschenbach J R,Zentek J. Cytotoxicity and metabolic stress induced by deoxynivalenol in the porcine intestinal IPEC-J2 cell line[J]. J Anim Physiol Anim Nutr,2012,96(4):709-716.
[19] Hamilton R M,Thompson B K,Trenholm H L,et al. Effects of feeding white Leghorn hens diets that contain deoxynivalenol(vomitoxin)-contaminated wheat[J]. Poult Sci,1985,64(10):1840-1852.
[20] Capaldo C T,Powell D N,Kalman D. Layered defense:how mucus and tight junctions seal the intestinal barrier[J]. J Mol Med,2017,95(9):927-934.
[21] Suzuki T. Regulation of intestinal epithelial permeability by tight junctions[J]. Cell Mol Life Sci,2013,70(4):631-659.
[22] Liao P,Liao M,Li L,et al. Effect of deoxynivalenol on apoptosis,barrier function,and expression levels of genes involved in nutrient transport,mitochondrial biogenesis and function in IPEC-J2 cells[J]. Toxicol Res,2017,6(6):866-877.
[23] Wirawan E,Berghe V T,Lippens S,et al. Autophagy:for better or for worse[J]. Cell Res,2012,22(1):43-61.
[24] Ge D,Gao J,Han L,et al. Novel effects of sphingosylphosphorylcholine on the apoptosis of breast cancer via autophagy/AKT/p38 and JNK signaling[J]. J Cell Physiol,2019,234(7):11451-11462.

相似文献/References:

[1]丁举静,何成华,刘俐君,等.脱氧雪腐镰刀菌烯醇对BHK-21细胞的线粒体膜电位及Bax和Bcl-2蛋白表达的影响[J].南京农业大学学报,2011,34(2):101.[doi:10.7685/j.issn.1000-2030.2011.02.018]
 DING Ju-jing,HE Cheng-hua,LIU Li-jun,et al.Effect of deoxynivalenol on the mitochondrial membrane potentials and Bax,Bcl-2 expression of BHK-21 cells[J].Journal of Nanjing Agricultural University,2011,34(4):101.[doi:10.7685/j.issn.1000-2030.2011.02.018]
[2]何成华,樊彦红,刘国芳,等.一株脱氧雪腐镰刀菌烯醇转化菌的分离、鉴定及其转化效果研究[J].南京农业大学学报,2009,32(4):143.[doi:10.7685/j.issn.1000-2030.2009.04.027]
 HE Cheng-hua,FAN Yan-hong,LIU Guo-fang,et al.Isolation and identification of a strain of Aspergillus tubingensis with deoxynivalenol biotransformation capability[J].Journal of Nanjing Agricultural University,2009,32(4):143.[doi:10.7685/j.issn.1000-2030.2009.04.027]
[3]耿芳芳,许伟,范梦雪,等.脱氧雪腐镰刀菌烯醇对雏鸡脂质过氧化反应及脑部形态结构的影响[J].南京农业大学学报,2016,39(3):460.[doi:10.7685/jnau.201510024]
 GENG Fangfang,XU Wei,FAN Mengxue,et al.Effects of deoxynivalenol on lipid peroxidation reaction and brain morphology in chicken[J].Journal of Nanjing Agricultural University,2016,39(4):460.[doi:10.7685/jnau.201510024]
[4]许伟,范梦雪,孟婷婷,等.脱氧雪腐镰刀菌烯醇对PC12细胞凋亡的影响[J].南京农业大学学报,2016,39(5):838.[doi:10.7685/jnau.201508040]
 XU Wei,FAN Mengxue,MENG Tingting,et al.Effect of deoxynivalenol on apoptosis of PC12 cells[J].Journal of Nanjing Agricultural University,2016,39(4):838.[doi:10.7685/jnau.201508040]
[5]王育伟,吴文达,施志玉,等.脱氧雪腐镰刀菌烯醇对大鼠胃液组成的影响[J].南京农业大学学报,2017,40(2):320.[doi:10.7685/jnau.201601015]
 WANG Yuwei,WU Wenda,SHI Zhiyu,et al.Effects of deoxynivalenol on the composition of gastric juice in rats[J].Journal of Nanjing Agricultural University,2017,40(4):320.[doi:10.7685/jnau.201601015]
[6]陈晓芳,张娅菲,储小燕,等.脱氧雪腐镰刀菌烯醇对仔猪海马神经细胞凋亡及线粒体膜电位的影响[J].南京农业大学学报,2018,41(3):526.[doi:10.7685/jnau.201705024]
 CHEN Xiaofang,ZHANG Yafei,CHU Xiaoyan,et al.Effects of deoxynivalenol on apoptosis and mitochondrial membrane potential in piglet hippocampal nerve cells[J].Journal of Nanjing Agricultural University,2018,41(4):526.[doi:10.7685/jnau.201705024]
[7]朱电锋,张娅菲,储小燕,等.脱氧雪腐镰刀菌烯醇对断奶仔猪生长性能、血清细胞因子及肠道免疫水平的影响[J].南京农业大学学报,2018,41(6):1126.[doi:10.7685/jnau.201711002]
 ZHU Dianfeng,ZHANG Yafei,CHU Xiaoyan,et al.Effects of deoxynivalenol on growth performance, serum cytokine and intestinal immune levels in weanling piglets[J].Journal of Nanjing Agricultural University,2018,41(4):1126.[doi:10.7685/jnau.201711002]
[8]储小燕,张娅菲,朱雷,等.自噬对DON诱导仔猪海马神经细胞损伤的影响[J].南京农业大学学报,2019,42(1):160.[doi:10.7685/jnau.201804028]
 CHU Xiaoyan,ZHANG Yafei,ZHU Lei,et al.Effect of autophagy on piglet hippocampal nerve cells damaged by deoxynivalenol[J].Journal of Nanjing Agricultural University,2019,42(4):160.[doi:10.7685/jnau.201804028]

备注/Memo

备注/Memo:
收稿日期:2019-01-26。
基金项目:国家自然科学基金项目(31772811);安徽省高校学科拔尖人才学术资助重点项目(gxbjZD2016120);安徽省高校自然科学研究重点项目(KJ2017A564);芜湖职业技术学院校级自然科学研究重点项目(Wzyzrzd201907)
作者简介:萧晟,副教授,兽医硕士,主要从事兽医临床研究,E-mail:xiaosheng28@126.com。
通信作者:黄克和,教授,主要从事动物营养代谢病和中毒病研究,E-mail:khhuang@nja
更新日期/Last Update: 1900-01-01