健步虎潜丸含药血清对体外成骨样细胞增殖和功能的影响

摘要

背景：寻找不仅能够抑制骨吸收，而且能够促进骨形成的补肾中药复方防治骨质疏松症是目前研究的热点。目的：探讨健步虎潜丸含药血清对体外成骨样细胞增殖及功能的影响。方法：24只成年雌性SD大鼠随机分为健步虎潜丸组低剂量组、中剂量组、高剂量组及生理盐水组，并分别以含生药1.5，3.0，6.0 g/kg健步虎潜丸及等体积生理盐水灌胃，每天2次，连续给药1周，末次给药后1 h处死制备含药血清。以各组血清分别处理成骨样细胞24，48，72 h，观察细胞形态，MTT法测定细胞增殖能力，碱性磷酸酶染色法检测碱性磷酸酶活性。结果与结论：①从24 h开始健步虎潜丸低剂量组、中剂量组及高剂量组的细胞密度高于对照组，核分裂相多见，细胞重叠生长，分泌物多，基质堆积明显，健步虎潜丸高剂量组细胞改变尤为显著；②各药物干预组较对照组吸光度值明显增加，差异有显著性意义。24 h时健步虎潜丸中、高剂量组较低剂量组吸光度值明显增加，差异有显著性意义。48，72 h时各药物干预组组间比较，差异有显著性意义；③各药物干预组较对照组碱性磷酸酶活性值明显增加，差异有显著性意义。各药物干预组组间比较，差异有显著性意义；④结果表明，健步虎潜丸含药血清能提高成骨样细胞的活性且存在浓度依赖性及时间依赖性。%BACKGROUND:More recently, the focus has been on searching for a compound Chinese medicine for reinforcing kidney, which cannot only inhibit bone absorption, but also promote osteogenesis to protect against osteoporosis. OBJECTIVE:To explore effects of drug serum of Jianbuhuqian pil s on proliferation and function of osteoblast-like cel s in vitro. METHODS:Twenty-four adult female Sprague-Dawley rats were randomly divided into low-dose, medium-dose, high-dose and normal saline groups, and given intragastric administration of 1.5, 3.0, and 6 g/kg Jianbuhuqian pills and equal volume of normal saline, respectively twice daily for 1 week. At 1 hour after final gavage, rats were decapitated to prepare drug sera used for culturing osteoblast-like cells. At 24, 48 and 72 hours of culture, the cellular morphology was observed, as well as the cell proliferation and alkaline phosphatase activity was detected by MTT assay and alkaline phosphatase staining, respectively. RESULTS AND CONCLUSION:Compared with the normal saline group, the cel density began to increase significantly in three Jianbuhuqian groups at 24 hours after culture, mitotic figures were easy to be observed, cel s were in overlapping growth, much secretions and matrix accumulation appeared, especial y in the high-dose group. The obsorbance values in Jianbuhuqian groups were significantly higher than that in the normal saline group. After 24 hours of culture, the obsorbance values in the medium-dose and high-dose groups were significantly increased compared with the low-dose group, and the values showed significant differences among three Jianbuhuqian groups after 48 and 72-hour culture. In addition, the alkaline phosphatase activity presented overt increase in the Jianbuhuqian groups compared with the normal saline group, and significant differences could be found among Jianbuhuqian groups. To conclude, the drug serum of Jianbuhuqian pil s can promote the activity of osteoblast-like cel s in a time-and concentration-dependent manner.