超临界流体萃取补肾活血方产物GC/MS分析及诱导成骨细胞增殖研究

GC/MS Characterization and Improvement of Osteoblast Cells Proliferation Study on Supercritical Fluid Extraction Products from Busheng Huoxue Formula

目的：研究超临界流体萃取选择性萃取补肾活血方产物的化学组成及体外诱导成骨细胞增殖的活性。方法：采用多元超临界流体萃取补肾活血方,采用气相色谱-质谱联用（GC/MS）表征各萃取产物的化学组成,并以体外培养原代大鼠颅成骨细胞为模型,研究各萃取产物含药血清诱导成骨细胞增殖的活性。结果：补肾活血方经超临界流体萃取得到SFEI和SFEII两个产物,GC/MS分析显示两个萃取产物挥发性组分的化学组成极为相似,为倍半萜和醛类化合物,但组成比例不同;体外研究显示SFEI与SFEII高、中、低剂量组含药血清均可显著诱导成骨细胞增殖（P〈0.05）,且SFEI中剂量组、SFEII低剂量组促成骨细胞增殖作用显著强于醇-水提取物组和阳性对照药组（P〈0.05）。结论：超临界流体可萃取选择性萃取补肾活血方中的挥发性和脂溶性成分,且该萃取物具有体外诱导成骨细胞增殖的活性。

Objective： Our study purposed to investigate the chemical composition of the products from Busheng Huoxue Formula,which was selectively extracted by the supercritical fluid extraction technology. We also examined the induction activity of the osteoblast cells’ proliferation in vitro. Methods： We employed multiply supercritical fluids to extract Busheng Huoxue Formula,and performed GC/MS to characterize the chemical composition of the SFE products. Then we investigated the proliferation activity of osteoblast cells from the skulls of newborn rats after treated with drug-loaded serums from each SFE product. Results： We extracted two products SFEI and SFEII from Busheng Huoxue Formula. The results from GC/MS analysis indicated that the main compounds in SFEI and SFEII were both sesquiterpenes and aldehydes. The both products shared the similar chemical compounds but distinguished from their composition ratio. Both drug-load serums from different SFEI and SFEII oral dose groups could significantly induce the proliferation of osteoblast cells in vitro,comparing with those of model group,positive drug group and Busheng Huoxue Formula ethanol-water extraction groups（ P 〈 0. 05）. Conclusion： SFE could selectively extract volatile and fat-soluble products from BushengHuoxue Formula,which could significantly induce the proliferation of osteoblast cells in vitro.