摘要
目的观察牙龈卟啉菌脂多糖(Pg-LPS)刺激的小鼠单核巨噬细胞RAW264.7培养上清对成骨细胞Runx2/Cbfα1表达的影响。方法收集经100 ng/ml Pg-LPS刺激的单核巨噬细胞RAW264.7培养24 h后上清,以20%的稀释浓度的分别作用于MC3T3-E1细胞1、6、12、24、48、72 h后,采用半定量RT-PCR与Western-Blot检测细胞Runx2/Cbfα1在基因与蛋白水平上的差异性表达。结果成骨细胞MC3T3-E1在20%稀释浓度的培养上清刺激后,半定量RT-PCR法发现Runx2/Cbfα1基因表达显著受抑制,且呈时间依赖性,72 h降到最低;Western-Blot检测显示细胞Runx2/Cbfαa1蛋白表达6 h后开始下降,72 h降至最低。结论 Pg-LPS刺激的小鼠单核巨噬细胞RAW264.7培养上清能抑制小鼠成骨细胞Runx2/Cbfα1的表达且与时间呈一定相关。
Objective To observe the expression of Runx2/Cbfα1 in osteoblasts cultivated in Pg-LPS-stimulated RAW264.7 culture supernatant.Methods100 ng/ml Pg-LPS was used to stimulate the murine monocyte-macrophage cells RAW264.7,24 h later,culture supernatant was collected and 20% concentration of which was applied to MC3T3-E1 cells for 1 h、6 h、12 h、24 h、48 h、72 h in vitro.The expression of Runx2/Cbfα1 mRNA in MC3T3-E1 cells was examined by semi-quantitive RT-PCR,and the expression of Runx2/Cbfα1 protein was tested by Western-blot.ResultsAfter cultivated in 20% concentration of Pg-LPS stimulated RAW264.7 culture supernatant,the expression of Runx2/Cbfα1 mRNA in MC3T3-E1 cells obviously decreased and had a negative correlation with time;the expression of Runx2/Cbfα1 protein in MC3T3-E1 cells decreased after 6 h,and after to the lowest point 72 h.ConclusionsPg-LPS-stimulated monocyte culture supernatant can suppress the expression of Runx2/Cbfa1 gene and Runx2/Cbfα1 protein in murine osteoblasts.
出处
《口腔医学》
CAS
2011年第3期140-143,共4页
Stomatology
基金
浙江省自然科学基金项目(Y207360)
