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目的 :观察经牙龈卟啉单胞菌脂多糖(Pg-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 OPG/RANKL表达的影响。方法:收集经100 ng/m L Pg-LPS刺激的单核巨噬细胞RAW264.7培养24 h后的上清,以不同稀释浓度(10%、20%、30%、40%和50%)的培养上清作用于MC3T3-E1 24h,分别通过RT-PCR、免疫印迹法(Western blotting)检测细胞OPG/RANKL基因和蛋白水平表达的变化,采用SPSS17.0软件包对数据进行单因素方差分析。结果:MC3T3-E1经不同稀释浓度的培养上清刺激后,其OPGm RNA及蛋白表达随浓度的增加而显著减少(P<0.05),而RANKLm RNA及蛋白表达随浓度的增加显著增加(P<0.05)。结论:Pg-LPS刺激的小鼠单核巨噬细胞RAW264.7培养上清可通过抑制成骨细胞OPGm RNA及蛋白的表达,增加RANKLm RNA及蛋白的表达而抑制其成骨能力,且与浓度呈一定的正相关。
OBJECTIVE: To observe the effect of RAW264.7 supernatant of mouse monocyte-macrophage stimulated by Pg-LPS on the expression of OPG / RANKL in mouse osteoblast MC3T3-E1. Methods: The supernatants of RAW264.7 monocytes stimulated with 100 ng / mL Pg-LPS for 24 h were collected and cultured in different dilutions (10%, 20%, 30%, 40% and 50% The expression of OPG / RANKL gene and protein were detected by RT-PCR and Western blotting respectively. The data were analyzed by one-way ANOVA with SPSS17.0 software package . Results: The mRNA and protein expressions of OPGm in MC3T3-E1 cells were significantly decreased (P <0.05), while the expression of RANKLm RNA and protein significantly increased with the increase in concentration (P < 0.05). CONCLUSION: The RAW264.7 culture supernatant of Pg-LPS-stimulated mouse monocyte-macrophage cells can inhibit the osteogenic capacity by inhibiting the expression of OPGm RNA and protein in osteoblasts and increasing the expression of RANKLm RNA and protein, There is a certain positive correlation.