LPS刺激单核巨噬细胞培养上清对成骨细胞OPG/RANKL的影响

上海口腔医学 ›› 2015, Vol. 24 ›› Issue (4): 428-432.

LPS刺激单核巨噬细胞培养上清对成骨细胞OPG/RANKL的影响

王海燕¹, 秦化祥¹, 邓辉², 孙超凡¹, 胡荣党¹

1.温州医科大学附属口腔医院正畸科,2.牙周病科,浙江温州 325027

收稿日期:2015-01-16 出版日期:2015-08-20 发布日期:2015-09-10
通讯作者: 胡荣党,Tel: 0577-88855488,E-mail: hurongdang@yahoo.com.cn E-mail:hurongdang@yahoo.com.cn
作者简介:王海燕（1983-）,女,硕士,住院医师,E-mail: 179402394@qq.com
基金资助:
浙江省自然科学基金（Y207360）; 温州市科技计划项目（Y20130259）

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

WANG Hai-yan¹, QIN Hua-xiang¹, DENG Hui², SUN Chao-fan¹, HU Rong-dang¹

1.Department of Orthodontics, 2.Department of Periodontics, School and Hospital of Stomatology, Wenzhou Medical University. Wenzhou 325027, Zhejiang Province, China

Received:2015-01-16 Online:2015-08-20 Published:2015-09-10
Supported by:
Supported by Natural Science Foundation of Zhejiang Province(Y207360) and Science and Technology Research Project of Wenzhou City (Y20130259)

摘要/Abstract

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/mL 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OPGmRNA及蛋白表达随浓度的增加而显著减少(P<0.05),而RANKLmRNA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OPGmRNA及蛋白的表达,增加RANKLmRNA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。

关键词: LPS, 单核巨噬细胞, 成骨细胞, OPG/RANKL

Abstract: 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 24h, then RT-PCR was used to detect the expression changes of OPG/RANKL mRNA.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 OPGmRNA and protein significantly decreased (P<0.05), whereas the expressions of RANKLmRNA 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 OPGmRNA and protein of osteoblastic cells, while increasing the expression of RANKLmRNA and protein in a concentration-dependent manner .

Key words: LPS, Monocyte, Osteoblasts, OPG/RANKL Shanghai J Stomatol, 2015, 24(4):428-432.

中图分类号:

R781.4

王海燕, 秦化祥, 邓辉, 孙超凡, 胡荣党. LPS刺激单核巨噬细胞培养上清对成骨细胞OPG/RANKL的影响[J]. 上海口腔医学, 2015, 24(4): 428-432.

WANG Hai-yan, QIN Hua-xiang, DENG Hui, SUN Chao-fan, HU Rong-dang. Effects of LPS-stimulated monocyte culture supernatant on the OPG/RANKL of osteoblastic cells[J]. Shanghai Journal of Stomatology, 2015, 24(4): 428-432.

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