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目的:研究补肾活血固齿方对小鼠颅顶前成骨细胞(MC3T3-E1细胞)BMP2表达及超微结构的影响,探讨该方剂治疗牙周炎的作用机理。方法:MC3T3-E1细胞分别于补肾活血固齿方含药血清、无药血清中培养24、48、72 h,ELISA法检测MC3T3-E1细胞BMP2的表达;培养14 d后,透射电镜观察MC3T3-E1细胞超微结构的变化。结果:含药血清组培养上清液的光密度值明显高于无药血清组,统计结果显示差异有显著性(P<0.05)。MC3T3-E1细胞经药物作用后呈现成骨细胞样表型,细胞表面突起变多,核卵圆形、较大,核膜凹陷,核仁明显;胞质丰富,粗面内质网扩张明显,网腔内充满低电子密度絮状物;高尔基复合体、线粒体发达,具有良好的的分泌功能,细胞代谢活跃。结论:补肾活血固齿方能促进MC3T3-E1细胞分泌BMP2,参与诱导成骨作用;补肾活血固齿方能诱导MC3T3-E1细胞呈成骨细胞样表型,促进成骨细胞的分化生长。
Objective: To study the effect of Bushen Huoxuesugu Recipe (BMPR) on BMP2 expression and ultrastructure in mouse cranial top osteoblasts (MC3T3-E1 cells) and to explore the mechanism of action of this recipe on periodontitis. Methods: The MC3T3-E1 cells were cultured for 24,48 and 72 h in Bushenhuoxuegucuangfang serum containing drug-free serum and the expression of BMP2 in MC3T3-E1 cells by ELISA. After cultured for 14 days, the expression of MC3T3- E1 cell ultrastructure changes. Results: The optical density of the culture supernatant of the drug-containing serum group was significantly higher than that of the drug-free serum group. The statistical results showed that the difference was significant (P <0.05). MC3T3-E1 cells showed osteoblast-like phenotype after drug treatment. The number of cell surface protrusions increased, the nucleus was oval and large, the nuclei were hollow and the nucleoli were obvious. The cytoplasm was rich and the rough endoplasmic reticulum was dilated obviously. Network cavity filled with low electron density floc; Golgi complex, developed mitochondria, has a good secretion, cell metabolism is active. Conclusion: Bushen Huoxuegucuang can promote the secretion of BMP2 in MC3T3-E1 cells, which is involved in the induction of osteogenesis. Bushenhuoxuegucuan can induce osteoblastic phenotype of MC3T3-E1 cells and promote the osteoblastic differentiation and growth.