LPS刺激单核巨噬细胞培养上清对成骨细胞OPG/RANKL的影响  

Effects of LPS-stimulated monocyte culture supernatant on the OPG/RANKL of osteoblastic cells

作　　者：王海燕[1] 秦化祥[1] 邓辉[2] 孙超凡[1] 胡荣党[1] 

机构地区：[1]温州医科大学附属口腔医院正畸科,浙江温州325027 [2]温州医科大学附属口腔医院牙周病科,浙江温州325027

出　　处：《上海口腔医学》2015年第4期428-432,共5页Shanghai Journal of Stomatology

基　　金：浙江省自然科学基金(Y207360);温州市科技计划项目(Y20130259)~~

摘　　要：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。PURPOSE：To investigate the influence of Pg-LPS stimulated monocyte（RAW264.7） culture supernatant on the OPG/RANKL expression of osteoblastic cells（MC3T3-E1）. METHODS： The culture supernatant of monocytes stimulated with Pg-LPS was applied to osteoblasts MC3T3-E1 with different diluted concentrations（10%, 20%, 30%, 40%and 50%） simultaneously for 24 h, then RT-PCR was used to detect the expression changes of OPG/RANKL m RNA.Western blot was used to detect the expression changes of OPG/RANKL protein. The data was analyzed by ANOVA using SPSS 17.0 software package. RESULTS：After stimulation of different concentrations of inflammatory supernatant, the expressions of OPGm RNA and protein significantly decreased（P 〈0.05）, whereas the expressions of RANKLm RNA and protein significantly increased（P〈0.05）, both of them were in a concentration-dependent manner. CONCLUSIONS：These results indicate that Pg-LPS stimulated RAW264.7 culture supernatant can inhibit the osteogenesis and differentiation of the osteoblasts through inhibiting the expression of OPGm RNA and protein of osteoblastic cells, while increasing the expression of RANKLm RNA and protein in a concentration-dependent manner.

关 键 词：LPS 单核巨噬细胞 成骨细胞 OPG/RANKL 

分 类 号：R78[医药卫生—口腔医学]