分子营养 Molecular Nutrition

呕吐毒素对IPEC-J2细胞凋亡的影响

展开
  • 1. 湖南中医药大学药学院, 长沙 410208;
    2. 中国科学院亚热带农业生态研究所, 亚热带农业生态过程重点实验室, 长沙 410125
廖美芳(1989-),女,湖南怀化人,硕士研究生,从事猪肠道细胞分子生物研究。E-mail:499136399@qq.com

收稿日期: 2017-09-01

  网络出版日期: 2018-03-05

基金资助

国家自然科学基金青年基金(31402088)

Effects of Deoxynivalenol on Apoptosis of IPEC-J2 Cells

Expand
  • 1. School of Pharmacy, Hunan University of Traditional Chinese Medicine, Changsha 410208, China;
    2. Key Laboratory for Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China

Received date: 2017-09-01

  Online published: 2018-03-05

摘要

本研究以IPEC-J2细胞为材料,研究不同浓度和时间处理下呕吐毒素(DON)对猪空肠上皮细胞凋亡的影响。通过CCK-8试剂盒检测细胞增殖毒性,筛选出合适的DON作用时间(6、12、24 h)。通过低浓度(200 ng/mL)和高浓度(2 000 ng/mL)DON分别作用IPEC-J2细胞6、12、24 h,检测DON对细胞有丝分裂周期和细胞早期凋亡的影响。结果表明:1)DON对IPEC-J2细胞生长有明显抑制作用,200和2 000 ng/mL DON浓度下,48和72 h的细胞存活率显著低于24 h(P<0.05);2 000 ng/mL DON浓度下,72 h的细胞存活率显著低于48 h(P<0.05)。2)不同DON浓度对细胞周期影响不同,低浓度DON主要作用于细胞有丝分裂S期,干扰DNA的复制;而高浓度DON主要作用于细胞有丝分裂G2/M期,干扰有关酶与纺锤丝蛋白质的合成。3)DON作用6 h,低浓度DON组和高浓度DON组早期凋亡率和总凋亡率显著高于对照组(P<0.05);DON作用12 h,高浓度DON组早期凋亡率和总凋亡率显著高于对照组和低浓度DON组(P<0.05)。由此可见,DON抑制了IPEC-J2细胞增殖,低浓度DON使细胞有丝分裂停留在S期,高浓度DON使细胞有丝分裂停留在G2/M期。DON促使细胞早期凋亡,使总凋亡率上升,且促凋亡作用随DON浓度升高而增强。

本文引用格式

廖美芳, 孟英才, 詹济华, 印遇龙, 廖鹏, 李玲 . 呕吐毒素对IPEC-J2细胞凋亡的影响[J]. 动物营养学报, 2018 , 30(3) : 1027 -1034 . DOI: 10.3969/j.issn.1006-267x.2018.03.027

Abstract

This experiment was conducted to determine the effect of deoxynivalenol (DON) on the apoptosis of IPEC-J2 cells after different concentrations and time treatment. An appropriate exposure time (6, 12, and 24 hour) used CCK-8 assay to test for cell proliferation. The effects of DON on cell mitosis cycle and early cell apoptosis were examined by treated the IPEC-J2 cells with low concentration (200 ng/mL) and high concentrations (2 000 ng/mL) DON for 6, 12, 24 hours. The results showed as follows:1) DON significantly inhibited the growth of IPEC-J2 cells. At 200 and 2 000 ng/mL DON concentrations, the cell survival rate of 48 and 72 h was significantly lower than that of 24 h (P<0.05), and the cell survival rate of 72 h was significantly lower than that of 48 at 2 000 ng/mL DON concentration (P<0.05). 2) Different DON concentrations had different effects on cell cycle. The low concentration of DON mainly acted on mitosis stage S and interferes with the replication of DNA, but the high concentration of DON mainly acted on the mitotic phase G2/M and interferes with the synthesis of enzymes and spindle filament proteins. 3) DON for 6 hours, the early apoptosis rate and total apoptosis rate of low concentration DON group and high concentration DON group were significantly higher than those of control group (P<0.05); DON for 12 hours, the early apoptosis rate and total apoptosis rate of high concentration DON group were significantly higher than those of control group and low concentration DON group (P<0.05). In conclusion, DON can inhibit IPEC-J2 cells proliferation, low concentration of DON can caused cell mitosis to stay in the S phase, high concentration of DON can caused cell mitosis to stay in the G2/M period. DON also can promote cell apoptosis and increase total apoptosis rate, and this effect is enhanced with DON concentration increasing.

参考文献

[1] PESTKA J J.Deoxynivalenol-induced proinflammatory gene expression:mechanisms and pathological sequelae[J].Toxins,2010,2(6):1300-1317.

[2] PESTKA J J,SMOLINSKI A T.Deoxynivalenol:toxicology and potential effects on humans[J].Journal of Toxicology and Environmental Health:Part B,2005,8(1):39-69.  

[3] YOSHIZAWA T,TAKEDA H,OHI T.Structure of a novel metabolite from deoxynivalenol,a tricho-thecene mycotoxin,in animals[J].Agricultural and Biological Chemistry,1983,47(9):2133-2135.

[4] COPPICK R W,SWANSON S P,GELBERG H B,et al.Preliminary study of the pharmacokinetics and toxicopathy of deoxynivalenol (vomitoxin) in swine[J].American Journal of Veterinary Research,1985,46(1):169-174.

[5] PESTKA J J.Deoxynivalenol:mechanisms of action,human exposure,and toxicological relevance[J].Archives of Toxicology,2010,84(9):663-679.  

[6] ZHANG H,VAN DER LEE T,WAALWIJK C,et al.Population analysis of the Fusarium graminearum species complex from wheat in China show a shift to more aggressive isolates[J].PLoS One,2012,7(2):e31722.

[7] PRELUSKY D B,VEIRA D M,TRENHOLM H L,et al.Excretion profiles of the mycotoxin deoxynivalenol,following oral and intravenous administration to sheep[J].Fundamental and Applied Toxicology,1986,6(2):356-363.  

[8] RIZZO A F,ATROSHI F,AHOTUPA M,et al.Protective Effect of Antioxidants against Free Radical-Mediated Lipid Peroxidation Induced by DON or T-2 Toxin[J].Transboundary and Emerging Diseases,1994,41(1/2/3/4/5/6/7/8/9/10):81-90.

[9] LAUTRAITE S,PARENT-MASSIN D,RIO B,et al.In vitro toxicity induced by deoxynivalenol (DON) on human and rat granulomonocytic progenitors[J].Cell Biology and Toxicology,1997,13(3):175-183.  

[10] CHUNG Y J,ZHOU H R,PESTKA J J.Transcriptional and posttranscriptional roles for p38 mitogen-activated protein kinase in upregulation of TNF-α expression by deoxynivalenol (vomitoxin)[J].Toxicology and Applied Pharmacology,2003,193(2):188-201.  

[11] 甄阳光,柏凡,张克英,等.我国主要饲料原料及产品中呕吐毒素污染分布规律研究[J].中国畜牧杂志,2009,45(8):21-24,28.

[12] VARGE E,MALACHOVA A,SCHWARTZ H,et al.Survey of deoxynivalenol and its conjugates deoxynivalenol-3-glucoside and 3-acetyl-deoxynivalenol in 374 beer samples[J].Food Additives & Contaminants:Part A,2013,30(1):137-146.  

[13] MARIANI V,PALERMO S,FIORENTINI S,et al.Gene expression study of two widely used pig intestinal epithelial cell lines:IPEC-J2 and IPEC-1[J].Veterinary Immunology and Immunopathology,2009,131(3/4):278-284.

[14] ALM H,GREISING T,BRVSSOW K P,et al.The influence of the mycotoxins deoxynivalenol and zearalenol on in vitro maturation of pig oocytes and in vitro culture of pig zygotes[J].Toxicology in Vitro,2002,16(6):643-648.  

[15] TIEMANN U,VIERGUTZ T,JONAS L,et al.Influence of the mycotoxins α-and β-zearalenol and deoxynivalenol on the cell cycle of cultured porcine endometrial cells[J].Reproductive Toxicology,2003,17(2):209-218.  

[16] DIESING A K,NOSSOL C,DANICKE S,et al.Vulnerability of polarised intestinal porcine epithelial cells to mycotoxin deoxynivalenol depends on the route of application[J].PLoS One,2011,6(2):e17472.

[17] DIESING A K,NOSSOL C,PANTHER P,et al.Mycotoxin deoxynivalenol (DON) mediates biphasic cellular response in intestinal porcine epithelial cell lines IPEC-1 and IPEC-J2[J].Toxicology Letters,2011,200(1/2):8-18.
文章导航

/