不同浓度脂多糖刺激奶牛乳腺上皮细胞对NF-κB及LAP表达的影响  

Effect of lipopolysaccharide with different concentrations on nuclear factor kappa B and LAP in dairy cow mammary epithelial cells

作　　者：程兰玲[1] 曹贵方[1] 温世勇[1] 赵鹏伟[1] 凃勇[1] 菅瑞珍[1] 李琦[1] 

机构地区：[1]内蒙古农业大学动物胚胎与发育生物学研究室,内蒙古呼和浩特010018

出　　处：《中国兽医科学》2012年第10期1068-1072,共5页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(30460093;31060328);内蒙古高等学校科学研究重点领域资助项目(ZL010144);转基因生物新品种培育重大专项(2008ZX08007-004)

摘　　要：为了观察不同浓度脂多糖对奶牛乳腺上皮细胞NF-κB及LAP基因表达的影响,设置不同浓度(0、50、100、200、400和800ng/mL)的脂多糖(LPS)刺激奶牛乳腺上皮细胞,分别于刺激2、4、8、16、24、48和72h后提取细胞总RNA,经反转录反应后,运用荧光定量PCR方法检测NF-κB P65亚单位和LAP的表达水平。结果,与空白对照组相比,脂多糖浓度达到400ng/mL并培养72h后,奶牛乳腺上皮细胞NF-κB P65亚单位和LAP的表达量最高,且与对照组有极显著差异(P<0.01)。结果表明,在脂多糖诱导的奶牛乳腺上皮细胞炎性效应中,NF-κB的表达活性增强,调控着防御基因LAP表达,呈现一定的剂量和时间效应关系。The objective was to observe the gene expression of nuclear factor kappa B（NF-κB） and lingual antimicrobial peptide（LAP） in dairy cow mammary epithelial cells with different dosages of lipopoltsaccharide（LPS） in vitro.The dairy cow mammary epithelial cells were stimulated with different concentrations（50,100,200,400 and 800 ng/mL） of LPS.The total RNA was extracted after stimulated for seven different time（2,4,8,16,24,48 and 72 h）,and the mRNA expression levels of NF-κB P65 and LAP were evaluated by real-time quantitative PCR.In result,the NF-κB P65 and LAP mRNA expression levels treated with 400 ng/mL LPS for 72 h,were the highest compared with the control group（P〈0.01）.The result confirmed that the expression activity of NF-κB was enhanced,and it regulated the expression of defense gene LAP with a certain dose and time effects.

关 键 词：Β-防御素 NF-ΚB 乳腺上皮细胞 脂多糖 

分 类 号：S852.161[农业科学—基础兽医学]