鸡胚肠上皮细胞Toll样受体的表达谱研究

›› 2013, Vol. 40 ›› Issue (11): 46-50.

鸡胚肠上皮细胞Toll样受体的表达谱研究

唐梦君^1,2, 高玉时^1,2, 张小燕^1,2, 陆俊贤^1,2, 蒲俊华^1,2, 顾荣^1,2

1. 农业部家禽品质监督检验测试中心, 江苏扬州 225125;
2. 中国农业科学院家禽研究所, 江苏扬州 225125

收稿日期:2013-05-02 出版日期:2013-11-20 发布日期:2013-12-19
通讯作者: 高玉时
作者简介:唐梦君(1980-),女,重庆人,博士,研究方向:食品安全检测与控制技术。
基金资助:
江苏省自然科学基金(BK2010297);江苏省家禽科学研究所博士基金(JQ201002)。

Study on Toll-like Receptors Expression Profile in Chicken Embryo Intestinal Epithelial Cells

TANG Meng-jun^1,2, GAO Yu-shi^1,2, ZHANG Xiao-yan^1,2, LU Jun-xian^1,2, PU Jun-hua^1,2, GU Rong^1,2

1. Quality Inspection Center for Poultry of Ministry of Agriculture, Yangzhou 225125, China;
2. Poultry Insitute, Chinese Academy of Agricultural Sciences, Yangzhou 225125, China

Received:2013-05-02 Online:2013-11-20 Published:2013-12-19

摘要/Abstract

摘要： 为明确鸡胚肠上皮细胞中Toll样受体(Toll-like receptors,TLR)的表达情况,本研究采用嗜热菌蛋白酶分离培养18日龄鸡胚的肠上皮细胞,在形态学观察及免疫细胞化学法鉴定基础上,采用RT-PCR方法检测其TLR表达谱,并对不同品种鸡胚肠上皮细胞TLR表达谱进行了比较研究。结果表明,应用嗜热菌蛋白酶消化法得到的鸡胚肠上皮细胞生长状况良好;采用低速离心和差速贴壁进行纯化,得到90%以上纯度的鸡胚肠上皮细胞;纯化后的细胞经形态学观察及细胞角蛋白单克隆抗体鉴定为阳性;10种TLR在鸡胚肠上皮细胞上均有mRNA表达,但其丰度存在明显差异;不同品种鸡胚肠上皮细胞TLR表达谱均无显著差异。

关键词: 鸡胚肠上皮细胞; Toll样受体; 表达谱

Abstract: Chicken epithelial cells (IECs) from eighteen day-old chicken embryos were separated by the digestion of thermolysin. The chicken epithelial cells were identified by morphological observation and immunocytochemical method. TLR mRNA expression was detected with reversing transcription polymerase chain reaction (RT-PCR). TLR receptor expression in the intestinal epithelial cells of different varieties of chick were compared. The results showed that the epithelial cells of chicken embryo grew well. 90% for the purification of the IEC was obtained by removing single cells by low-speed centrifuge and differential velocity adherent. The result of identification by detecting the cyto-keratin was positive. IECs all expressed of 10 kinds of TLR mRNA with different intensity. There was no significant difference in TLR receptor expression in the intestinal epithelial cells of different varieties of chick.

Key words: chicken embryo intestinal epithelial cells; Toll-like receptor; expression profile

中图分类号:

S831

唐梦君, 高玉时, 张小燕, 陆俊贤, 蒲俊华, 顾荣. 鸡胚肠上皮细胞Toll样受体的表达谱研究[J]. , 2013, 40(11): 46-50.

TANG Meng-jun, GAO Yu-shi, ZHANG Xiao-yan, LU Jun-xian, PU Jun-hua, GU Rong. Study on Toll-like Receptors Expression Profile in Chicken Embryo Intestinal Epithelial Cells[J]. , 2013, 40(11): 46-50.

参考文献

1 王晓亮,顾宪红,郑冬梅,等.鸡小肠上皮细胞的分离及体外培养研究进展[J].中国畜牧兽医,2010,37(9):34~38.
2 张道杰,蒋建新,陈永华,等.用嗜热菌蛋白酶进行人肠上皮细胞分离培养[J].第三军医大学学报,2004,26(11):1016~1018.
3 Abreu M T,Fukata M,Arditi M. TLR signaling in the gut in health and disease[J].J Immunol,2007,19(2):106~115.
4 Aldhous M C,Shamkov A N,Bode J,et al. Characterization of conditions for the primary culture of human small intestinal epithelial cells[J]. Clin Exp Immunol,2001,125(1):32~40.
5 Bouhet S,Hourcade E,Loiseau N,et al. The mycotoxin fumonisin B₁ alters the proliferation and the barrier function of porcine intestinal epithelial cells[J]. Toxicol Sci,2004,77(1):165~170.
6 Greenman H S,Rutten M J,Fowler W M,et al. Herbicide/Pesticide effects on intestinal epithelial growth[J]. Environ Res,1997,75:85~93.
7 He Y P,Chu S W,Walker W A. Nucleotide supplements later proliferation and differentiation of cultures human (Caco-2) and rat (IEC-6) intestinal epithelia cells[J]. J Nutr,1993,123:1017~1027.
8 Iqbal M,Philbin V J,Smith A L. Expression patterns of chicken Toll-like receptor mRNA in tissues,immune cell subsets and cell lines[J]. Vet Immunol Immunopathol,2005,104(1~2):117~127.
9 Kang S S,Kaul L S,Gaspari A A. Toll-like receptors applications to dermatologic disease[J]. J Am Acad Dermatol,2006,54(6):951~983.
10 Kedinger M,Assmann P,Alexandre E,et al. Importance of a fibroblastic support of in vitro differentiation of intestinal endodermal cells and for their response to glucorticoids[J]. Cell Differ,1987,20(3):171~182.
11 Ortega-Cava C F,Ishihara S,Rumi M A,et al. Strategic compartment alization of Toll-like receptor 4 in the mouse gut[J]. J Immunol,2003,170(8):3977~3985.
12 Oswald I P. Role of intestinal epithelial cells in the innate immune defence of the pig intestine[J]. Vet Res,2006,37(3):359~368.
13 Sanderson I R,He Y P. Nucleotide uptake and metabolism by intestinal epithelial cells[J].J Nutr,1994,124:131~137.
14 Temperley N D,Berlin S,Paton I R,et al. Evolution of the chicken Toll-like receptor gene family: A story of gene gain and gene loss[J].BMC Genomics,2008,9:62.
15 Uehara A,Fujimoto Y,Fukase K,et al. Various human epithelial cells express functional Toll-like receptors,NOD1 and NOD2 to produce anti-microbial peptides,but not proinflammatory cytokines[J]. Mol Immunol,2007,44(12):3100~3111.

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