健骨颗粒含药血清对成骨样细胞增殖及PCNA、Cyclin D1 mRNA表达的影响

The Effect of Jiangu Granula on the Proliferation and PCNA、Cyclin D1 Genetic Expression in Osteoblast-like Cell in Vitro

目的:观察健骨颗粒含药血清对成骨样细胞增殖及PCNA、Cyclin D1 mRNA表达的影响。方法:制备健骨颗粒含药血清和生理盐水血清,分别干预体外培养的成骨样细胞ROS17/2.8,MTT法检测细胞增殖,流氏细胞术检测细胞周期,荧光定量PCR检测PCNA、Cyclin D1 mRNA表达。结果:与同浓度生理盐水血清组相比,中药血清组10%-30%浓度促细胞增殖均有显著性差异(P<0.05),其中浓度为20%时,两组间有非常显著性差异(P<0.01);中药血清组G0/G1期细胞比例明显低于同期生理盐水血清组,有非常显著性差异(P<0.01)。G2/M期、S期细胞比例及PI在干预48h时均高于同期生理盐水血清组,具非常显著性差异(P<0.01)。中药血清组较理盐水血清组PCNA、Cyclin D1 mRNA表达升高,组间有显著性差异(P<0.05)。结论:健骨颗粒含药血清能促进成骨样细胞的增殖、提高PCNA、Cyclin D1 mRNA表达。

Objective ：To observate the effect of Jiangu granula on the proliferation and PCNA、Cyclin D1 Genetic expression in Osteoblast-like cell in vitro.Method： Preparation for serum containing Jiangu Granula and normal saline（NS）,intervene in Osteoblast-like cell ROS17/2.8 for 48h and detect the proliferation by MTT method.Analyzed the cell cycle by flow cytometry（FCM） and calculated cells proliferation index（PI）.To detect the expression levels of PCNA、Cyclin D1 mRNA by SYBR GREEN real-time polymerase chain reaction.Result： Rat of cell proliferation was the fastest when the concentration of serum containing Jiangu Granula was 20%.It was significantly faster than NS-group with the concentration between 10% and 30%.（P〈0.05）.The cell percentage of G0/G1 phase of TCM-group was lower than NS-group significantly（P〈0.01）.While the percentage of G2/M、S phase and PI were higher than NS-group significatantly with the same time（P〈0.01）.The expression level of PCNA and Cyclin D1 mRNA of TCM-Group was higher than NS-group（P〈0.05）.Conclusion：The serum containing Jiangu Granula stimulated OB in vitro to proliferate obviously,promoted the expression level of PCNA and Cyclin D1 mRNA.