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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 effects of Erigeron breviscapus (L.) Breviscapus (Hymenoptera: Erigerontis Breviscapus) on the expression of osteopontin (OPG) and RANKL protein in cultured osteoblasts and the mechanism of action of Erigeron Breviscapus on orthodontic bone remodeling. METHODS: Osteoblast-like cell line MG63 cells were cultured in vitro. The cell mechanical tension was stimulated by SXG4201 four-point bending cell mechanical loader. The effects of mechanical tension tension (2000 μstrain, 0.5 Hz), single breviscapus (1 mg/mL) intervention, and mechanical tension (2000 μstrain, 0.5 Hz) and breviscapus (1 mg/mL) alone on MG63 cells were different. After time (3h, 6h, 12h, 24h), total cellular RNA and protein were extracted, and the expression of OPG protein and RANKL protein in MG63 cells was detected by Western blot. Results: The expression of OPG protein was up-regulated after SMA stimulation with mechanical tension, and it was time-dependent, but there was no significant change in RANKL protein expression. OPG protein expression was down-regulated and RANKL protein expression was increased after breviscapine intervention alone in MG63 cells. After combined intervention of MG-63 cells, the expression of OPG protein decreased and the expression of RANKL protein increased significantly. Conclusion: Erigeron breviscapus can antagonize the stimulatory effect of mechanical tension on OPG secretion in osteoblasts and promote the stimulatory effect of mechanical tension on RANKL secretion in osteoblasts.