作　　者： (段荟芹）; (王利）; (李键）; (钟金城）; (字向东）; (江明锋）; (熊显荣）;

机构地区： 西南民族大学青藏高原研究院,四川成都610041

出　　处： 《中国兽医学报》 2017年第9期1743-1747,共5页

摘　　要： 采用PCR方法从牦牛成纤维细胞中扩增TLR4基因,并用5mg/L质量浓度的LPS诱导牦牛成纤维细胞,分别在0,12,24,36h用实时荧光定量PCR检测TLR4及其信号通路基因(IL-1β、IL-8)mRNA水平的相对表达量。结果显示,克隆得到TLR4基因序列长2 067bp(GenBank号:KU743904),与GenBank中野牦牛TLR4的相似性达99%。5mg/L质量浓度的LPS诱导处理后的各基因与0h相比,在12和24h均有上调,在36h均下调。诱导后24h,TLR4、IL-1β和IL-8基因的表达量均达到最大值,且都显著高于0和36h(P<0.05)。由此得出TLR4、IL-1β和IL-8均可在牦牛皮肤成纤维细胞中表达,并参与了对LPS的应答反应。 TLR4 gene was cloned from yak fibroblasts by RT-PCR. Yak fibroblasts were exposed to 5 mg/L LPS for 0,12,24 and 36 h. The expression level of TLR4,IL-8,IL-1β were detected by re- al-time PCR. The results showed that the nucleotide sequence of TLR4 gene was 2 067 bp,which of GenBank accession number was KU743904. This gene showed 99% homology with Bos taurus and other species. The expression level of all genes increased at 12 and 24 h,while all genes mRNA levels decreased at 36 h. The TLR4,IL-1β and IL-8 mRNA levels reached a maximum value in yak skin fibroblasts after stimulation with 5 mg/L LPS for 24 h,which were up-regulated significantly compared with 0 and 36 h （P〈0.05）. The rerults indicated that TLR4,IL-8 and IL-1β can be ex- pressed in yak skin fibroblasts. They were all involved in the responses to LPS.

关 键 词： 成纤维细胞 牦牛 信号通路 细胞因子