牙龈卟啉单胞菌脂多糖(Pg-LPS)对破骨细胞EphA2基因表达的调控

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (3): 268-271.

牙龈卟啉单胞菌脂多糖(Pg-LPS)对破骨细胞EphA2基因表达的调控

高爱超¹, 于海燕¹, 李娜¹, 侯玉帛¹, 戚欣², 于小琳³, 于维先^1,3*

1. 吉林大学口腔医学院牙周病科吉林长春 130021;
2. 北华大学;
3. 吉林大学口腔医学院,吉林省牙发育及颌骨重塑与再生重点实验室

收稿日期:2014-07-03 出版日期:2015-03-28 发布日期:2016-04-29
通讯作者: 于维先,电话:0431-88796000
作者简介:高爱超(1987~ )女,山东人,硕士,医师,主要从事口腔牙周病学临床和基础研究工作。
基金资助:
吉林省科技厅自然科学基金资助课题(编号:201115104)

Effect of Porphyromonas Gingivalis Lipopolysaccharide (Pg-LPS) on the Expression of EphA2 Gene in Osteoclast.

GAO Ai-chao, YU Hai-yan, LI Na, et al.

Department of Periodontics, the Stomatological Hospital of Jilin University, Changchun 130021

Received:2014-07-03 Online:2015-03-28 Published:2016-04-29

摘要/Abstract

摘要： 目的:研究小鼠单核巨噬细胞系RAW264.7在破骨分化过程中,Pg-LPS对破骨细胞EphA2表达的影响。方法:用终浓度10 mg/L的Pg-LPS 刺激RAW264.7 细胞后,分别在1、3、5 d,应用RT-PCR检测破骨细胞中EphA2基因和破骨细胞相关基因(破骨细胞内基质金属蛋白酶( MMP9)、ACP5、c-fos、组织蛋白酶K(CtsK)、NFATc1)的表达,并且通过酒石酸抗酸性磷酸酶(TRAP)染色观察实验组和对照组破骨细胞的分化成熟情况。结果:RT-PCR检测10 mg/L的Pg-LPS在第3天和5天,实验组比对照组EphA2基因表达分别增高2.4倍和1.2倍,两组之间存在显著差异(P<0.01);同时也能够促进破骨相关基因c-fos、NFATc1、CtsK、ACP5、MMP9的表达,实验组与对照组相比差异有显著性;TRAP染色结果显示:实验组比对照组的TRAP阳性多核细胞数目明显增多。结论:10 mg/L的Pg-LPS对小鼠单核巨噬细胞系RAW264.7,在破骨分化的中期和晚期均能够促进EphA2基因的表达,但是在破骨分化早期对EphA2基因的表达无明显作用。

关键词: Pg-LPS, 破骨细胞, EphA2基因

Abstract: Objective: To investigate the effect of porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on the expression of EphA2 gene in RAW264.7 cells during osteoclastic differentiation. Methods: RAW264.7 cells were stimulated with 10μg/ml of porphyromonas gingivalis lipopolysaccharide (Pg-LPS) at 1, 3 and 5 d. RT-PCR was applied to determine the expression of EphA2 gene and the osteoclast related genes(MMP9,c-fos,ACP5,CtsK,NFATc1). Tartrate-resistant acid phosphatase (TRAP) staining was applied to observe osteoclast differentiation and maturation. Results: Compared with the control group, at the 3 d and 5 d, the EphA2 gene mRNA expression was significantly increased 2.4-fold and 1.2-fold in Pg-LPS group. At the 1 d, there was no obvious difference between the Pg-LPS group and control group. Meanwhile Pg-LPS stimulation significantly promote osteoclast related genes of c-fos, NFATc1, CtsK, ACP5 and MMP9 expressions.Tartrate-resistant acid phosphatase (TRAP) staining showed that compared with the control group, the number of the TRAP positive cells was significantly increased in the Pg-LPS group. Conclusion: Pg-LPS can promote the expression of EphA2 gene in the middle and later stages of osteoclast differentiation,but has no obvious effect in the early stage.

Key words: RAW264.7 , Pg-LPS, EphA2 gene

中图分类号:

R781.4

高爱超, 于海燕, 李娜, 侯玉帛, 戚欣, 于小琳, 于维先. 牙龈卟啉单胞菌脂多糖(Pg-LPS)对破骨细胞EphA2基因表达的调控[J]. 口腔医学研究, 2015, 31(3): 268-271.

GAO Ai-chao, YU Hai-yan, LI Na, et al.. Effect of Porphyromonas Gingivalis Lipopolysaccharide (Pg-LPS) on the Expression of EphA2 Gene in Osteoclast.[J]. Journal of Oral Science Research, 2015, 31(3): 268-271.

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