为了探明NLRP3在鸡小肠上皮细胞中表达情况,为鸡的NLRP3基因在小肠细胞中的功能研究奠定基础。采用18胚龄的鸡胚,分离培养鸡的小肠上皮细胞(intestinal epithelial cells,IEC);采用免疫细胞化学(immunocytochemistry,ICC)和反转录PCR(RT-PCR)方法,检测NLRP3基因在鸡小肠上皮细胞中的表达。结果表明:分离培养出活性较强的IEC;ICC检测显示获得的小肠上皮细胞表面NLRP3抗原阳性,RT-PCR法可扩增出520 bp的目的片段。表明成功分离培养出鸡小肠上皮细胞,并证明鸡NLRP3在鸡小肠上皮细胞中表达。
The aim of this study is to ascertain the expression of NLRP3 in intestinal epithelial cells(IECs)of chicken,andlay the foundation for the functional studies of chicken NLRP3 gene in IECs. Intestinal epithelial cells were isolated and culturedfrom chicken embryo at 18 days. The expression of chicken NLRP3 was detected by immunocytochemistry(ICC)and reverse tran-scription PCR(RT-PCR)method.The results showed that chicken IEC grew very well.The surface antigen of NLRP3 was positiveby ICC,and the 520 bp fragments of NLRP3 gene was obtained by RT-PCR length.Chicken IECs were isolated and cultured suc-cessfully.These results indicate that chicken NLRP3 gene exists in chicken IECs.