不同炮制品入药的青娥丸含药血清对人成骨细胞增殖、分化及矿化的影响

目的:研究以不同炮制品入药的青娥丸含药血清对人成骨细胞增殖、分化和矿化的影响。观察盐炙品及生品青娥丸含药血清对人成骨细胞功能的影响,探讨青娥丸对骨质疏松症的治疗机制,阐述应用盐炙品配伍入药的必要性。方法:以绝经后妇女股骨松质骨分离得到的成骨细胞为研究对象,制备以补骨脂和杜仲生品及盐炙品配伍入药的青娥丸,给予大鼠生品及盐炙品青娥丸水提液ig(10mL·kg-1),给药持续7d,第7天采集大鼠血液制备含药血清。实验分为空白组、空白血清组、青娥丸含药血清组、盐青娥丸含药血清组。采用MTT法检测各组含药血清对人成骨细胞增殖的影响;同时测量成骨细胞碱性磷酸酶活性来评价各组含药血清对人成骨细胞分化的作用;采用细胞钙茜素红染色方法对给予含药血清处理的人成骨细胞进行染色,观察对人成骨细胞矿化的影响。结果:与同浓度空白血清比较,青娥丸的生品和盐炙品的含药血清均有促进成骨细胞增殖的作用(P<0.05),且含药血清浓度相同时,青娥丸盐炙品含药血清促进成骨细胞增殖的作用优于生品含药血清(P<0.05);青娥丸生品及盐炙品含药血清具有促进成骨细胞分化的作用(P<0.05),且盐炙品的作用要优于生品(P<0.01);空白血清对成骨细胞的矿化无显著的作用,而青娥丸生品及盐炙品含药血清具有促进成骨细胞矿化结节形成的作用,且盐炙品的作用要优于生品。结论:青娥丸能提高成骨细胞增殖、分化及矿化活性,其对于骨质疏松症的治疗作用与其能显著提高成骨细胞功能有关,且盐炙品入药的青娥丸效果较好。

Serum Containing Drug of Different Processed Products of Qing' e Pill on Osteoblast Proliferation, Differentiation and Mineralization

Objective: To study the serum containing drug of different processed products of Qing'e Pill on osteoblast proliferation, differentiation and mineralization. This study was designed to evaluate the promoting effect of serum containing drug of different processed products of Qing'e Pill on human osteoblast and investigate the possible mechanism in the treatment of osteoporosis. Method: Osteogenic cells were obtained from the femur in postmenopausal women. Different processed products of Qing'e Pill were performed by using crude and salt-roast Eucommia ulmoides and Psoralea corylifolia, respectively. Female Sprague Dawley rats were orally administrated with water extract of Qing'e Pill for 7 days. At the end of the administration, serum containing drug was collected. The experiment was divided into the blank group, the blank serum group, the different processed products of Qing'e Pill groups. Cell proliferation was determined by MTT assay. Cell (activity of alkaline phosphatase) ALP activity and mineralized nodule were tested using ELISA. Result: Compared to the blank serum group, the different processed products of Qing'e pill could significantly promote huaman osteoblast proliferation and ALP activity (P<0.05). Moreover, the formation of bone nodules increased in the osteoblasts after 14-day treatment in different processed products of Qing'e pill groups (P<0.05). Meanwhile, the ALP (P<0.01) and mineralization activity results of the salt-roast were better than the crude. Conclusion: Qing'e pill could increase proliferation, differentiation and mineralization activities in human osteoblast. Its therapeutic effect of osteoporosis might be involved in increasing the osteogenic function. Comparing to the crude drug, the salt-roast has a better effect in treating osteoporosis.