阳和汤体外对大鼠成骨细胞及破骨细胞增殖的影响

Effects of Yanghe Decoction Containing Serum on Osteoblasts and Osteoclast in Rats

目的探讨阳和汤含药血清对成骨细胞、破骨细胞的影响作用。方法 SD大鼠连续7 d灌胃阳和汤,腹主动脉采血,制备含药血清。分离并培养原代成骨细胞、成骨细胞株UMR106、原代破骨细胞、破骨前体细胞RAW264.7。随机分为阳和汤含药血清低剂量组、中剂量、高剂量,空白血清组,各组加入相应含药血清进行培养,分别于24、48、72h收获细胞,采用MTT法测定药物对细胞增殖的影响;比色法测定破骨细胞TRAP酶活性。结果阳和汤含药血清中剂量、高剂量组在24、48、72h均能提高成骨细胞的增殖率,与空白血清组比较有显著差异;阳和汤含药血清低剂量组在48、72h也能明显提高细胞增殖率,与空白血清组比较有显著差异;阳和汤含药血清低剂量组对破骨前体细胞RAW264.7增殖率24、48、72h均没有影响,与空白血清组比较差异均无统计学意义;阳和汤含药血清中、高剂量组破骨前体细胞RAW264.7增殖率均低于空白血清组,24、48、72h比较差异有统计学意义;阳和汤含药血清低剂量组对破骨细胞的TRAP活性24、48 h影响不大,但72h TRAP活性低于空白血清组;阳和汤含药血清中、高剂量组48、72h破骨细胞TRAP活性均显著低于空白血清组。结论阳和汤能抑制体外培养的破骨前体细胞增殖,降低破骨前体细胞TRAP活性,对成骨细胞的增殖有促进作用。

Objective： To study the function of the medicine-containing serum of Yanghe Decoction effect on the osteoblast and osteoclast of rats. Methods： Rats were fed with Yanghe Decoction once a day for 7 days,the rats＇blood was collected and separated to obtain the medicine-containing serum. The generation of osteoblasts, os- teogenesis cell lines of UMR106 and precursor cells of osteoclast RAW264.7 were randomly divided into low dose group,middle dose group,high dose group and blank serum control group of the medicine-containing serum of Yanghe Decoction. Each group joined the corresponding medicated serum training and harvest cells at different time period as 24 h,48 h,and 72 h. And we used MTT method to determine the effect of drugs on cell prolifera- tion. The colorimetric method to determine the broken bone cells RAP enzymes. Results： The serum contained high dose and medium dose of Yanghe Decoction could significantly improve osteoblast proliferation rate at 24 h, 48 h, as well as 72 h. While low dose could obviously promote osteoblast proliferation rate only at 48 h and 72 h. Comparison with the control, these differences had notably statistical significance. The medicine-containing serum of Yanghe Decoction low dose group was without the influence of precursor ceils of osteoclast RAW264.7 in 24 h, 48 h and 72 h. No statistically significant differences were compared with the blank group. The proliferation rates of precursor cells of osteoclast RAW264.7 in middle dose group and high dose group were lower than the blank serum control group. These differences had notably statistical signifieances in 24 h,48 h and 72 h. Different dosage of serum containing Yanghe Decoction could not obviously prohibit the differentiation of osteoclasts through the investigation of TRAP test. Conclusion： The medicine containing serum of Yanghe Decoction can in- hibit proliferation of the precursor cells of osteoclast culture in vitro, reduce the TRAP active of precursor cells of osteoclast and improve the generation of osteoblasts.