灯盏花联合机械牵张力对成骨细胞OPG和RANKL蛋白表达的影响

Effects of erigeron breviscapus and mechanical force on the expression of OPG and RANKL in osteoblasts in vitro

目的:研究灯盏花、机械牵张力以及灯盏花与机械牵张力联合作用下对体外培养的成骨细胞OPG和RANKL蛋白表达的影响,探讨灯盏花对正畸骨改建的作用机理。方法:体外培养成骨样细胞株MG63细胞,细胞的机械牵张力刺激采用SXG4201型四点弯曲细胞力学加载仪。分别单用机械牵张力刺激(2000 μstrain,0.5Hz)、单用灯盏花(1mg/mL)干预、以及机械牵张力(2000 μstrain,0.5Hz)和灯盏花(1mg/mL)联合干预MG63细胞不同时间(3h,6h,12h,24h)后,提取细胞总RNA和蛋白质,用Western blot方法检测MG63细胞OPG蛋白和RANKL蛋白的表达。结果:单用机械牵张力刺激MG63细胞后OPG蛋白表达上调,且呈时间依赖性,而RANKL蛋白表达无明显变化;单用灯盏花干预MG63细胞后OPG蛋白表达下调,RANKL蛋白表达增加;两者联合干预MG-63细胞后,OPG蛋白表达量减少,RANKL蛋白表达显著增加。结论:灯盏花能拮抗机械牵张力对成骨细胞OPG分泌的刺激作用,促进机械牵张力对成骨细胞RANKL分泌的刺激作用。

Objective：To study the effection of erigeron breviscapus （EB） and （or） mechanical force on the expression of OPG and RANKL protein in MG-63 osteoblast-like cells in vitro, thus to propose the role that EB can play in orthodontic bone rebuilding. Metbod：MG-63 osteoblast-like cells were cultured in vitro,and were stimulated by a mechanical instrument called SXG4201 model. The 3th passage of cells were divided into the control group and the different experiment group.The different groups were intervented respectively by EB （1 mg/ mL）, mechanical force （2000 μstrain, 0.5 Hz） ,EB （1 mg/mL） combined with mechanical force （2000 μstrain,0.5 Hz）. Total RNA and protein were respectively isolated from cells after 3,6, 12 and 24 hours. Expressions of OPG protein and RANKL protein in MG-63 ostcoblast-like cells were detected by Western blot. Result：Mechanical force （2000 μstrain,0.5 Hz） increased the expression of OPG protein in MG-63 osteoblast-like cells,but could not change the expression of RANKL protein in MG-63 osteoblast-like cells. From 3,6, 12 to 24 hours after intervented by mechanical force,the expression of OPG protein in MG-63 osteoblast- like cells was increased in time dependent. EB （1 mg / mL） decreased the expression of OPG protein in MG-63 osteoblast- like cells, and increased the expression of RANKL protein in MG-63 osteoblast-like cells. EB （1 mg / mL） combined with mechanical force （2000 μstrain,0.5 Hz）decreased the expression of OPG protein in MG-63 osteoblast-like cells,and increased apparently the expression of RANKL protein in MG-63 osteoblast-like cells. Conclusion：EB could inhibit osteoblasts the secretion of OPG induced by mechanical force,and could promote osteoblasts the secretion of RANKL induced by mechanical force.