大肠杆菌感染诱导的小尾寒羊外周血白细胞TLR1～5基因差异表达
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摘要
小尾寒羊经乳房实验性感染大肠杆菌,利用半定量RT-PCR方法检测大肠杆菌感染后不同时间小尾寒羊外周血白细胞中TLR1~5基因mRNA的表达情况。结果显示,大肠杆菌感染后外周血白细胞中TLR1~5基因mRNA的表达均出现先升高后降低的趋势。其中TLR3、TLR4和TLR5基因的相对表达量于大肠杆菌感染后24,36,48h均出现显著或极显著升高,而TLR2和TLR4基因的相对表达量于感染后72h时出现显著降低。结果表明,大肠杆菌感染后,TLR3、TLR4和TLR5受体与细菌的识别、炎症的发生密切相关,为进一步研究TLR1~5基因在绵羊大肠杆菌乳房炎发病过程中的作用机制奠定了基础。
Small-tail Han sheep was experimentally infected with E.coli through beast and semiquantitative RT-PCR method was adopted to detect the expression of mRNA of peripheral blood leukocytes after the infection at different times.The results showed that the expression of TLR1-5 mRNA of E.coli-affected peripheral blood leukocytes increased first and then decreased.The relative expression of TLR3,TLR4,TLR5 genes increased significantly or extremely significantly after24,36,48 hand TLR2,TLR4 gene relative expression decreased markedly after 72 h.This reveals that TLR3,TLR4,TLR5 receptors closely relate to bacteria identification and inflammation after the E.coli infection.This study lays the groundwork for the future research of TLR1-5 in the pathogenesis of E.coli mastitis.
Small-tail Han sheep was experimentally infected with E.coli through beast and semiquantitative RT-PCR method was adopted to detect the expression of mRNA of peripheral blood leukocytes after the infection at different times.The results showed that the expression of TLR1-5 mRNA of E.coli-affected peripheral blood leukocytes increased first and then decreased.The relative expression of TLR3,TLR4,TLR5 genes increased significantly or extremely significantly after24,36,48 hand TLR2,TLR4 gene relative expression decreased markedly after 72 h.This reveals that TLR3,TLR4,TLR5 receptors closely relate to bacteria identification and inflammation after the E.coli infection.This study lays the groundwork for the future research of TLR1-5 in the pathogenesis of E.coli mastitis.
引文
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[14]GALLI R,PAONE A,FABBRI M,et al.Toll-like receptor 3(TLR3)activation induces microRNA-dependent;reexpression of functional RAR beta and tumor regression[J].Proc Natl Acad Sci USA,2013,110(24):9812-9817.
[15]SALAUN B,ZITVOGEL L,ASSELINPATUREL C,et al.TLR3as a biomarker for the therapeutic efficacy of double-stranded RNA in breast cancer[J].Cancer Res,2011,71(5):1607-1614.
[16]侯一峰,何韶衡.革兰氏阳性菌肽聚糖对人嗜碱性粒细胞中TLR_3、TLR_9 mRNA的上调作用[J].汕头大学医学院学报,2005,18(2):74-76.
[2]王爱华,付茂华,程叶.布尔山羊乳腺炎病原菌的分离与鉴定[J].动物医学进展,2002,23(4):78-80.
[3]MEDZHITOV R,PRESTON-HURLBURT P,JA-NEWAY C A Jr.A human homologue of the Drosophila Toll protein signals activation of adaptive immunity[J].Nature,1997,388(6640):394-397.
[4]MATZINGER P.The danger model:a renewed sense of self[J].Science,2002,296(5566):301-305.
[5]BEUTLER B,JIANG Z,GEORGEL P,et al.Genetic analysis of host re-sistance;Toll-like receptor signaling and immuniy at large[J].Annu Rev Immunol,2006,24:353-389.
[6]IBEAGHAAWEMU E M,LEE J W,IBEAGHA AE,et al.Bacterial lipopolysaccharide induces increased expression of toll-like receptor(TLR)4and downstream TLR signaling molecules in bovine mammary epithelial cells[J].Vet Res,2008,39(2):11.
[7]GILBERT F B,CUNHA P,JENSEN K,et al.Differential response of bovine mammary epithelial cells to Staphylococcus aureus[J].Vet Res,2013,44(1):1-23.
[8]MINTZ M,MINTZ D,EZRA-ELIA R,et al.Pam3CSK4/TLR2signaling elicits neutrophil recruitment and restricts invasion of Escherichia coli P4into mammary gland epithelial cells in a murine mastitis model[J].Vet Immunol Immunopathol,2013,152(1/2):168-175.
[9]WU J,LI L,SUN Y,et al.Altered molecular expression of the TLR4/NF-κB signaling pathway in mammary tissue of Chinese Holstein cattle with mastitis[J].PLoS One,2015,10(2):e0118458.
[10]PORCHERIE A,CUNHA P,TROTEREAU A,et al.Repertoire of Escherichia coli agonists sensed by innate immunity receptors of the bovine udderand mammary epithelial cell[J].Vet Res,2012,14.doi:10.1186/1297-9716-43-14.
[11]MIAO E A,ANDERSEN-NISSEN E,WARREN SE,et al.TLR5 and Ipaf:dual sensors of bacterial flagellin in the innate immune system[J].Semin Immunopathol,2007,29:275-288.
[12]CHANG G,PETZL W,VANSELOW J,et al.Epigenetic mechanisms contribute to enhanced expression of immune response genes in the liver of cows after experimentally induced Escherichia coli,mastitis[J].Vet J,2014,203(3):339-341.
[13]MATSUMOTO M,SEYA T.TLR3:Interferon induction by double-stranded RNA including poly(I:C)[J].Adv Drug Delivery Rev,2008,60(7):805-812.
[14]GALLI R,PAONE A,FABBRI M,et al.Toll-like receptor 3(TLR3)activation induces microRNA-dependent;reexpression of functional RAR beta and tumor regression[J].Proc Natl Acad Sci USA,2013,110(24):9812-9817.
[15]SALAUN B,ZITVOGEL L,ASSELINPATUREL C,et al.TLR3as a biomarker for the therapeutic efficacy of double-stranded RNA in breast cancer[J].Cancer Res,2011,71(5):1607-1614.
[16]侯一峰,何韶衡.革兰氏阳性菌肽聚糖对人嗜碱性粒细胞中TLR_3、TLR_9 mRNA的上调作用[J].汕头大学医学院学报,2005,18(2):74-76.