摘要
通过研究脂多糖(LPS)诱导奶牛子宫内膜上皮细胞核因子κB亚单位P65蛋白mRNA的表达变化,从细胞水平深入探讨子宫内膜炎的发病机理。将奶牛子宫内膜上皮细胞传代培养,采用1、10、50、100、1000μg/mL五个浓度梯度的LPS刺激子宫内膜上皮细胞,MTT法筛选最佳刺激浓度;以上述最佳刺激浓度刺激细胞,于0、0.5、1、2、4h后收集细胞,荧光定量RT-PCR检测p65 mRNA的表达差异性。结果显示,100μg/mLLPS为最佳刺激浓度;LPS刺激1h组p65 mRNA的表达极显著(P<0.01)高于其他时间组;2h组显著(P<0.05)高于0、0.5和4h组;0.5和4h组显著(P<0.05)高于0h组。结果表明,LPS可诱导奶牛子宫内膜上皮细胞NF-κB的激活,LPS介导的NF-κB信号通路存在于奶牛子宫内膜上皮细胞中,并参与子宫内膜炎发病机理的调控。
To investigate the mechanism of endometritis in cellular level,we studied the changes of mRNA level of NF-κB subunit P65,which were induced by LPS in bovine endometrial epithelial cell.The bovine endometrial epithelial cells were subcultivated and stimulated by LPS at five different concentrations of 1,10,50,100 and 1 000μg/mL,respectively.The optimal concentration was selected by MTT.Stimulated with the optimal concentration for 0,0.5,1,2 or 4 hseparately,the cells were collected to detect the expression of p65 mRNA by RT-PCR.In results,the optimal concentration of LPS was 100μg/mL.The mRNA expression level of p65 in the 1 hgroup was extremely higher than that of the other groups（P0.01）. The mRNA expression level was higher in the 2hgroup compared with the 0,0.5 and 4 hgroups（P 0.05）.In the 0.5and the 4hgroups,the mRNA expression level was higher than the 0h group（P0.05）. In conclusion,NF-κB could be activated by LPS in bovine endometrial epithelial cell,and the NF-κB signal pathway could be induced by LPS in bovine endometrial epithelial cell and participated in the regulation mechanism of endometritis.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2010年第10期1063-1066,共4页
Chinese Veterinary Science
基金
山东省教育厅高等学校科技计划项目(J10LC03)
