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目的:在右归饮对体外正常大鼠成骨细胞的增殖和分化作用研究基础上,进一步观察右归饮对激素性股骨头坏死大鼠体外诱导培育成骨细胞的增殖和分化以及基因表达的影响。方法:取体重为100～120g雄性SD大鼠20只,臀肌注射醋酸强的松龙49mg/kg·d,连用6d(造模),7d后处死,无菌条件下取造模大鼠骨髓基质细胞,诱导培养成骨细胞,然后随机分为4组:高、中、低浓度右归饮含药血清组(R1、R2、R3)和对照组(R4),分别添加高、中、低浓度右归饮含药血清及对照血清,继续培养72h后,进行成骨细胞增殖、碱性磷酸酶活性、骨保护素(OPG)和破骨细胞分化因子(RANKL)表达水平检测(实时荧光定量PCR检测),并进行统计学分析。结果:高、中浓度右归饮含药血清组(R1、R2)与对照组(R4)相比,成骨细胞增殖率显著升高(P<0.01),碱性磷酸酶活性显著升高(P<0.01),成骨细胞OPGmRNA的相对表达量显著升高(P<0.01),且与浓度剂量呈一定的正相关;成骨细胞RANKLmRNA显著降低(P<0.01)。低浓度右归饮含药血清组(R3)与对照组(R4)相比,成骨细胞增殖率、碱性磷酸酶活性以及OPG、RANKLmRNA相对表达量无统计学差异。结论:高、中浓度的右归饮含药血清对体外培养的SINFH大鼠成骨细胞的增殖和分化具有明显的促进作用,并可提高成骨细胞OPGmRNA的相对表达量而对其RANKLmRNA有抑制作用。右归饮含药血清对体外诱导培养成骨细胞的作用与剂量有一定相关性,在一定的浓度范围内促进成骨细胞的分化、增殖。
OBJECTIVE: To study the effect of Youguiyin on the proliferation and differentiation of osteoblasts induced by steroid-induced osteonecrosis of the femoral head in vitro and on the basis of the study of the effect of Yougui Yin on the proliferation and differentiation of normal rat osteoblasts in vitro influences. METHODS: Twenty male SD rats weighing 100-120g were injected with glucocorticoid prednisolone 49mg / kg · d into the gluteal muscles for 6 days (model). After 7 days, the rats were sacrificed and the bone marrow The osteoblasts were induced and then divided into 4 groups at random: high, medium and low concentrations of Youguiyin containing serum group (R1, R2, R3) and control group (R4), respectively, high, medium and low Concentration of Yougui Yin containing serum and control serum, continue to culture 72h after osteoblast proliferation, alkaline phosphatase activity, osteoprotegerin (OPG) and osteoclast differentiation factor (RANKL) expression levels were detected (real-time fluorescence quantitative PCR detection), and statistical analysis. Results: Compared with the control group (R4), the proliferation rate of osteoblasts increased significantly (P <0.01) and the activity of alkaline phosphatase increased remarkably (P <0.01) (P <0.01). The relative expression of OPG mRNA in osteoblasts was significantly increased (P <0.01), and positively correlated with the concentration of OPG mRNA. The expression of RANKL mRNA in osteoblasts decreased significantly (P <0.01). Compared with the control group (R4), the osteoblast proliferation rate, alkaline phosphatase activity, and the relative expression levels of OPG and RANKL mRNA in the low concentration YUGUYINYU containing serum group (R3) were not significantly different. CONCLUSION: The high and medium concentrations of YGW drink serum can significantly promote the proliferation and differentiation of osteoblasts cultured in vitro and increase the relative expression of OPG mRNA in osteoblasts, while inhibiting the expression of RANKLmRNA effect. Youguiyin containing serum on osteoblasts in vitro induction of the role and dose have a certain relevance, in a certain concentration range to promote osteoblast differentiation and proliferation.