含骨碎补总黄酮血清对新生SD大鼠成骨细胞增殖、分化和矿化的影响

目的 探讨骨碎补总黄酮含药血清对新生SD大鼠成骨细胞增殖、分化、矿化的影响及其作用机制.方法 (1)骨碎补总黄酮含药血清的制备:选用12月龄雌性SD大鼠,药物灌胃,每日1次,连续12 d,末次灌药1.5 h后腹主动脉取血,分离血清备用.(2)实验模型的建立:采用二次酶消化法从新生SD大鼠颅骨获取成骨细胞,传代培养,取第3代成骨细胞为实验模型.(3)指标测定:①细胞增殖:用MTr法;②细胞分化:碱性磷酸酶(ALP)活性(PNPP法);骨钙素含量(放射免疫分析法);③细胞矿化:采用茜素红染色法,在40倍光镜下计矿化结节.结果 骨碎补总黄酮含药血清可刺激体外培养新生SD大鼠成骨细胞增殖,促进成骨细胞合成分泌ALP,且能增加成骨细胞骨钙素的分泌,促进矿化结节的形成.结论 骨碎补总黄酮可促进成骨细胞的增殖、分化和矿化,从而促进骨形成.

Effect of serum carrying Fortune's drynaria rhizome chromocor on proliferation, differentiation, and mineralization of osteoblasts of new-born SD rats

Objective To study the effect and mechanism of serum carrying Fortune's drynaria rhizome chomocor on proliferation, differentiation, and mineralization of osteoblast of new-born SD rats. Methods (1) Preparation of serum carrying Fortune's drynaria rhizome chromocor: Fortune's drynaria rhizome chromocor was administered once everyday for continuous 12 days to 12 months old female SD rats. Blood was obtained from aorta. (2) Creation of the experiment model; Isolated the osteoblast from the neonatal SD rats' calvaria by using enzyme digestion twice, subcultured. The third passage of cultured osteoblast was chosen as the experiment model. (3) Index; ①The proliferation capacity of osteoblast: detected by method of MTT. ②The differentiation activity of osteoblast: detected by the alkaline phosphatase assay (PNPP) and the amount of ostecalcin (radioimmunoassay). ③The mineralization activity of osteoblast: detected by counting mineral deposites under 40× microscope after chinalizarin staining. Results Serum carrying Fortune's drynaria rhizome chromocor stimulated proliferation of osteoblast of new-bom SD rats in vitro, promoted the excretion of alkaline phosphatase and osteocalsin, and increased the formation of the mineral deposites. Conclusions Fortune's drynaria rhizome chromocor can stimulate proliferation, differentiation, and mineralization of osteoblasts and promote the bone formation.