补骨脂单体成分对体外培养成骨细胞和破骨细胞分化的影响

The effects of psoralen components on mice osteoblast and osteoclast differention in vitro

关键词:  异补骨脂素  异补骨脂查尔酮  成骨细胞  破骨细胞  增殖  分化

Key words:  isopsoralen  isobavachalcone  osteoclasts  osteoblast  proliferation  differention

摘要:

[目的] 研究补骨脂单体成分异补骨脂素、异补骨脂查尔酮对体外培养破骨细胞抗酒石酸酸性磷酸酶(TRACP)及成熟破骨细胞形成数目、体外培养成骨细胞增殖与分化的影响。[方法] 体外采用巨噬细胞集落刺激因子(M-CSF)和核因子κB受体活化因子配体(RANKL)诱导培养破骨细胞前体细胞,考察异补骨脂素、异补骨脂素查尔酮对破骨细胞前体细胞在分化过程中其标志性酶TRACP的活性和成熟破骨细胞产生数目的影响。对硝基苯磷酸盐法(PNPP法)测定破骨细胞TRACP活性,TRACP细胞染色法鉴定并检测成数破骨细胞数目。酶消化法体外培养新生小鼠成骨细胞,(CCK-8)(cell counting kit-8)法检测两成分对成骨细胞增殖的影响,磷酸苯二钠法测定两成分对成骨细胞标志性酶碱性磷酸酶(ALP)的影响。[结果] 体外成功培养小鼠破骨细胞和成骨细胞,两细胞强表达各自的标志性酶TRACP和ALP.异补骨脂查尔酮在0.05 μmol/L浓度下对成熟破骨细胞的形成有显着抑制作用。异补骨脂素在0.1 μmol/L浓度下对成骨细胞增殖有显着促进作用。[结论] 补骨脂单体成分异补骨脂素对成骨细胞有促进作用、异补骨脂查尔酮对破骨细胞有抑制作用,提示补骨脂可能作为抗骨质疏松或骨吸收的药物。

Abstract:

[Objective] To study the effects of psoralen components, isopsoralen and isobavachalcone on the cultured osteoclasts TRACP enzyme activity and differention. Moreover, the effect of two components on the mice osteoblasts proliferation and differentiation were also studied. [Methods] Using M-CSF and RANKL, osteoclast precursor cells were induced to differentiated into mature osteoclasts. During the differention period, isopsoralen and isobavachalcone were added to cells, therefore PNPP were used to detected osteoclasts TRACP activity. ICC staining of TRACP were performed to identify mature osteoclasts number. The neonatal mice osetoblast cells were cultured in vitro, CCK-8 methods were adopted to detected the osteoblast proliferation properties. Sodium phenylphosphate were used to measure the osteoblast marker enzyme ALP activity. [Results] Mouse osteoclasts and osteoblasts were cultured successfully in vitro. Isobavachalcone significantly inhibit the mature osteoclast formation at the dose of 0.05 μM, and isopsoralen had a significant role in promoting osteoblast proliferation at the dose of 0.1 μM. [Conclusion] Psoralea components isopsoralen and isobavachalcone have effects on bone cells, suggesting that Fructus Psoraleae maybe act as a drug for anti-osteoporosis or anti-bone resorption.