健骨颗粒对成骨细胞分化的影响

目的:观察健骨颗粒对成骨细胞分化过程的影响。方法:取第3代SD大鼠颅骨成骨细胞,生理盐水血清组加入生理盐水血清;含药血清组加入最佳浓度的健骨颗粒颗粒含药血清;MTT法检测最佳含药血清浓度;运用采用ELISA法检测成骨细胞OCN的表达,采用比色法测羟脯氨酸(HYP)、碱性磷酸酶(AKP),实时荧光定量PCR-SYBR GREEN法检测核心结合因子(Cbfα1)、I型胶原(Col I)、成骨转录因子(OSX)mRNA的表达。取第4代细胞采用茜素红染色法染色矿化结节。结果:MTT法检测显示含药血清最佳浓度为20%;钙化结节观察显示含药20%组最先出现矿化结节,结节数量增多;含药血清组细胞液内OCN、AKP和HYP含量高于生理盐水血清组(P0.05);Real-time PCR检测结果显示含药血清组Cbfα1、ColⅠ、OSX基因mRNA的相对表达水平明显高于生理盐水血清组(P0.05)。结论:健骨颗粒含药血清能提高体外培养成骨细胞中矿化结节的数量、AKP、HYP、OCN的含量、ColⅠ、Cbfα1、OSX mRNA的表达,促进成骨细胞增殖,加快细胞分化。

Effect of Jiangu granula on osteoblast in differentiation

Objective: To observate the effect of Jiangu granula(JGG) on osteoblast in differentiation.Methods: Establishing the culture in vitro system of osteoblast: Primary cells were isolated from calvaria of newborn SD rats by enzymatic digestion.Cells at the third passage were selected for experiment.Detective index: Cell proliferation of osteoblast cultured in differenent concentration serum was measured by MTT Method.The expression of OCN was measured by ELISA.The expression of AKP and HYP was measured by color method.The expression of Cbfα1,OSX,ColⅠ was measured by SYBR GREEN.The 4th generation of cell is used the sodium alizarinsulfonate dyeing mineralization tubercle continuously.Results: MTT showed that cell proliferation rate of osteoblast was the fastest when the concentration of the serum containing JGG was 20%.The calcification tubercle was presented in the group including the medicine JGG 20%.The OCN,AKP and HYP content of the NS-group and the JGG-group increased along with the number of days,and the JGG-group was higher than the NS-group,two groups quite even had the significance difference(P<0.05).Realtime-PCR showed Cbfα1,ColⅠand OSX in osteoblast of JGG-group was significantly higher than that in NS-group(P<0.05).Conclusion: JGG can increase the expression of OCN,OSX,Cbfα1,AKP,ColⅠ,HYP and Mineralization tubercle,so that to multiply osteoblast,to speed up the cell differentiation.