脂多糖刺激的单核巨噬细胞培养上清液对小鼠MC3T3-E1细胞c-fos表达的影响

目的：研究经牙龈卟啉单胞菌（Porphyromonas gingivalis,P.g）脂多糖（LPS）刺激的小鼠单核巨噬细胞RAW264.7培养上清对小鼠成骨细胞（MC3T3-E1细胞）c-fos表达的影响。方法：收集100μg/L的Pg LPS刺激RAW264.724h后的上清液,以20%稀释浓度分别作用于MC3T3-E1,分别在1、3、6、12、24、48、72h时,采用RT-PCR和Western blot检测ALP mRNA、c-fos mRNA及蛋白表达水平的变化。结果：Pg LPS刺激RAW264.7细胞培养上清作用于MC3T3-E1后,ALP mRNA表达均下降,在6h时表达最低,而c-fos的mRNA和蛋白水平均提高,分别在3h和6h时达最高水平。结论：Pg LPS刺激的小鼠单核巨噬细胞RAW264.7培养上清可通过上调成骨细胞c-fos的表达而抑制其成骨能力。

Expression of c-fos in MC3T3-E1 after Affected by LPS Stimulated RAW264.7 Culture Supernatant

Objective：To investigate the expression of c-fos in MC3T3-E1 after affected by Pg LPS stimulated RAW264.7 culture supernatant. Methods：We selected the supernatant of the RAW264.7 which was stimulated by 100ng/ml Pg LPS. After put the supernate of 20% concentration into the cells of MC3T3-E1 for 1h,3 h,6h,12h,24h,48h,72h,we used the RT-PCR to analyze the gene of ALP and c-fos,and used the Western blot to analyze the protein of c-fos. Results：In the MC3T3-E1,affected by the supernate,the expression of ALP mRNA was decreased and reached the lowest point at 6h. But the expression of c-fos mRNA and protein were increased and reached the highest point at 3h and 6h respectively（P0.05） . Conclusion：The supernate fluid can up-regulate the expression of the gene of c-fos and the c-fos protein to inhibit the osteogenesis and differentiation of the osteoblasts.