BMP-2对体外培养人牙囊细胞表达OPG和RANKL的影响(1)
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[摘要]目的:研究BMP-2对体外培养人牙囊细胞表达OPG和RANKL的影响。方法:第5代人牙囊细胞免疫组化染色,检测人牙囊细胞中OPG和RANKL蛋白的表达;第5代人牙囊细胞与浓度为100 ng/ml的BMP-2共同孵育0h、1h、3h、6h、12h、18h,RT-PCR法检测OPG和RANKL基因表达的变化。结果:人牙囊细胞OPG、RANKL免疫组化染色阳性;100 ng/ml的BMP-2上调OPG蛋白的分泌,最佳效应时间为12～18h,下调RANKL基因的表达,最佳效应时间为6～12h。结论:人牙囊细胞存在OPG、RANKL蛋白的表达;100ng/ml的BMP-2可增强人牙囊细胞OPG蛋白分泌和基因表达,减弱RANKL基因的表达,降低RANKL/OPG的比值,抑制破骨细胞的形成。
[关键词]骨形成蛋白-2;人牙囊细胞;骨保护素;破骨细胞分化因子
[中图分类号]Q813.1[文献标识码]A [文章编号]1008-6455(2010)09-1313-04
Effects of BMP-2 on the expressions of OPG and RANK in the human dental follicle cells
SUN Hai-yan1,ZHANG Yong-kuan2,JIN Zuo-lin3,LIN Zhu3
(1.Department of Stomatology,the 307th Hospital of PLA,Beijing 100071,China; 2.Department of Stomatology,the 150th Hospital of PLA,Luoyang 471031,Henan,China; 3.Department of Orthodontics,the Fourth Millitary Medical University,Xi'an 710032,Shaanxi,China)
Objective:To study the effects of BMP-2 on the expressions of OPG and RANKL in the human dental follicle cells.MethodsThe expressions of OPG and RANKL were examined in the 5th passage human dental follicle cells by immunohistochemical staining technique.The human dental follicle cells were co-cultured with 100 ng/ml BMP-2 for 0,1h,3h,6h,12h,18h,then the expressions of OPG and RANKL were assayed by Reverse transcription polymerase chain reaction(RT-PCR). ResultsThe immunohistochemical staining of OPG and RANKL was positive. 100ng/ml BMP-2 up regulated the expression of OPG and the optimal time was 12h～18h,down regulated the expression of RANKL and the optimal time was 6h～12h.ConclusionsOPG and RANKL are expressed in human dental follicle cells. 100ng/ml BMP-2 up regulate the expression of OPG and down regulate the expression of RANKL.The decrease of RANKL/OPG inhibits osteoclastogenesis.
Key words: BMP-2;OPG;human dental follicle cells;RANKL
牙囊组织起源于外胚间充质,是包绕成釉器周围的疏松结缔组织,在牙齿萌出和牙周组织形成中起重要作用,缺乏牙囊的牙齿不能萌出,而单核细胞在牙囊的聚集以及单核细胞分化成破骨细胞、进而吸收牙槽骨、形成牙齿萌出通道是牙齿萌出的关键。这个过程受许多细胞因子的调控,如集落刺激因子-1(CSF-1)、甲状旁腺相关蛋白(PTHrP)、骨保护素(OPG)、破骨细胞分化因子(RANKL)等,这些因子直接或者间接诱导单核细胞进入牙囊。而牙齿萌出的过程伴随萌出牙齿冠方、根方牙槽骨的代谢,成骨与破骨在骨代谢中是密不可分的,骨形成蛋白-2(BMP-2)作为一种重要的成骨相关蛋白,在这个过程中又是如何发挥作用的呢? ......
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