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10mg/L的Pg-LPS刺激RAW264.7细胞后,分别在1、3、5d,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10mg/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:10mg/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。
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 10t^g/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 （MMPg, c -- fos, ACP5, CtsK,NFATcl）. Tartrate--resistant acid phosphatase （TRAP） staining was applied to observe osteoclast differen- tiation and maturation. Results： Compared with the control group, at the 3 d and 5 d, the EphA2 gene mRNA ex- pression was significantly increased 2.4--fold and 1.2--fold in Pg--LPS group. At the I 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--los, NFATcl, CtsK, ACP5 and MMP9 expressions. Tartrate--resistant acid phos- phatase （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.
出处
《口腔医学研究》
CAS
CSCD
北大核心
2015年第3期268-271,275,共5页
Journal of Oral Science Research
基金
吉林省科技厅自然科学基金资助课题(编号:201115104)
