续骨补肾方含药血清对成骨细胞增殖及骨形成蛋白-2,Ⅰ型胶原表达的影响

Effect of Serum Containing Xugu Bushen Recipe on Proliferation of Osteoblasts and the mRNA Expression of BMP-2 and Collagen Type Ⅰ

目的:探究续骨补肾方含药血清在骨折愈合过程中对成骨细胞增殖及BMP-2,Ⅰ型胶原mRNA表达的影响,进一步揭示其促进骨折愈合的具体机制,为临床推广提供实验依据。方法:分离培养大鼠成骨细胞并进行鉴定,传代备用并制备15%含药血清并分组(高、中、低剂量3个含药血清组和生理盐水组)+DMEM培养基;MTT法观察各组对成骨细胞增殖的影响;RT-PCR检测成骨细胞的Ⅰ型胶原、BMP-2 mRNA表达。结果:续骨补肾方含药血清组在24 h,48 h,72 h均可促进成骨细胞增殖,且在24 h低剂量组促进成骨细胞增殖作用最强;在72 h时,中剂量组促进成骨细胞增殖作用最强,差异有统计学意义。续骨补肾方含药血清各剂量组均可上调BMP-2,Ⅰ型胶原mRNA的表达(P<0.05),且低剂量组作用最强。结论:续骨补肾方可促进成骨细胞增殖,增加BMP-2,Ⅰ型胶原mRNA的表达,促进骨折愈合,且低剂量是最佳剂量。

Objective:To investigate the effect of serum containing Xugu Bushen recipe on the proliferation of osteoblasts and mRNA expression of BMP-2 and collagentypeⅠin the process of fracture healing.Methods:Isolation,culture and identification of osteoblasts;Preparation and grouping of drug-containing serums(high,medium and low dose serums and normal saline groups);MTT assay was used to observe the effect of each group on osteoblast proliferation;RT-PCR detection of osteoblast collagen typeⅠ,BMP-2 mRNA expression.Results:The serum group containing the prescription of Xugu Bushen decoction could promote the proliferation of osteoblasts at 24 h,48 h and 72 h.At 72 h,the medium dose group had the strongest effect on promoting the proliferation of osteoblasts.Xugu Bushen decoction drug-containing serum high and low dose group can raise BMP-2,collagen typeⅠmRNA expression(P<0.05),the strongest and low dose group.Conclusion:Xugu Bushen decoction can promote the proliferation of osteoblasts and increase the mRNA expression of collagen typeⅠand BMP-2.