牵张应力下骨碎补总黄酮对成骨细胞增殖及相关蛋白含量的影响

Effect of Total Flavonoids in Rhizoma Drynariae under Tensile Stress on Osteoblast Proliferation and Correlative Protein Content

目的:观察骨碎补总黄酮对牵张应力下成骨细胞增殖及相关蛋白含量的影响。方法:对大鼠成骨样细胞系ROS1728作体外培养,取3代对数生长期成骨细胞,采用Flexcell细胞牵张系统构建成骨细胞牵张体系,使用骨碎补总黄酮含药血清进行干预24 h。应用MTT法绘制成骨细胞生长曲线;碱性磷酸酶染色法观察成骨细胞内碱性磷酸酶(ALP)含量及分布情况;采用碱性磷酸酶定量检测方法测定各组成骨细胞内ALP含量;采用ELISA法测定骨形态发生蛋白-2(BMP-2)、转化生长因子-β1(TGFβ-1)的表达。结果:12 h及24 h骨碎补总黄酮各剂量组中成骨细胞数量及ALP、BMP-2、TGFβ-1含量均明显增加,与空白组比较,差异均有统计学意义(P<0.05);同时,与骨碎补总黄酮低剂量组比较,骨碎补总黄酮中、高剂量组成骨细胞数目及ALP和BMP-2的含量增高,差异均有统计学意义(P<0.05);骨碎补总黄酮各剂量组中TGFβ-1的含量比较,差异无统计学意义(P>0.05)。结论:在牵张应力下,骨碎补总黄酮可明显促进成骨细胞增殖及相关蛋白的合成。

Objective：To observe effect of total flavonoids in rhizoma drynariae under tensile stress on osteoblast proliferation and correlative protein content.Methods：Cultured rats＇ osteoblast-like cells lines ROS1728 in vitro.Get 3rd osteoblasts in the logarithmic phase and establish osteoblasts tensile system using Flexcell,intervene with total flavonoids in rhizoma drynariae drug serum for 24 h.Drew osteoblasts growth curve by method of MTT,and observed contents and distribution of alkaline phosphatase（ALP） in osteoblasts by alkaline phosphatase staining method,detected ALP contents of osteoblasts in every group by alkaline phosphatase quantitative test method,detected expression of bone morphogenetic protein-2（BMP-2） and transforming growth factor（TGFp-1） by ELISA method.Results：The number of osteoblasts and content of ALP,BMP-2 and TGFp-1 in all dose groups of 12 h and 24 h total flavonoids in rhizoma drynariae were all increased,comparing with those in blank group,differences being significant（P〈0.05）.At the same time,comparing with low dose group of total flavonoids in rhizoma drynariae,the number of osteoblasts and content of ALP and BMP-2 in mid and high dose group of total flavonoids in rhizoma drynariae were increased,differences being significant（P〈0.05）.Comparing content of TGFp-1 in all dose group of total flavonoids in rhizoma,difference was not significant（P〈0.05）.Conclusion：Total flavonoids in rhizoma drynariae can promote osteoblast proliferation and synthesis of involved protein obviously.