壮药扶芳藤提取物调控骨代谢的体外实验研究

In-vitro Experimental Study on Zhuang Medicine Fufangteng Extract Regulating Bone Metabolism

目的:观察壮药扶芳藤的高浓度提取物对体外培养的破骨细胞及成骨细胞骨代谢的影响。方法:采用大鼠颅盖骨机械法体外分离培养获得成骨细胞(osteoblast,OB);新生大鼠长干骨机械分离法获得破骨细胞(osteoclast,OC)。观察组培养液为10%胎牛血清+最低基本培养基+高浓度的扶芳藤提取物(扶芳藤乙醇提取物浸膏含量为200 mg/kg),对照组不加扶芳藤提取物。分别用四甲基偶氮唑盐和碱性磷酸酶法检测OB的增殖和分化状态;抗酒石酸酸性磷酸酶染色观察OC形态变化,图像分析骨吸收陷窝面积,原子分光光度计法测定OC培养液上清液中钙离子含量。结果:高浓度扶芳藤提取物可以提高OB的碱性磷酸酶(alkaline phosphatase,ALP)的活性,且能够造成体外培养的OC空泡性变,骨吸收陷窝面积减少和降低培养液上清液中钙离子含量。结论:壮药扶芳藤能够发挥类雌激素样作用,抑制骨吸收,减少骨丢失,间接促进骨形成,改善骨代谢,防治原发性I型骨质疏松症。

Objective:To observe the effects of high concentration extract of Fufangteng on bone metabolism of osteoclasts(OC)and osteoblasts(OB)cultured in vitro.Methods:OBs were isolated and cultured in vitro by mechanical method of rat calvaria;OCs were gained by mechanical separation of long stem bone in neonatal rats.The observation group used 10%fetal bovine serum+minimum essential medium+high concentration of Fufangteng extract(the contents of ethanol extract from Fufangteng were 200 mg/kg)as culture medium,and the control group didn′t use Fufangteng extract.MTT and ALP were respectively used to detect the proliferation and differentiation of OBs;tartrate resistant acid phosphatase staining(TRAP)to observe the morphological changes of OC,image analysis to test bone resorption lacuna area,and atomic spectrophotometry to determine calcium in supernatant of OC culture solution.Results:High concentration extract of Fufangteng could raise ALP activity of OB,cause OC vacuolation,decrease the area of bone resorption lacuna and lower the content of calcium in the supernatant of culture medium.Conclusion:Zhuang medicine Fufangteng could play an estrogen like role,inhibit bone resorption,reduce bone loss,promote bone formation indirectly,improve bone metabolism,prevent and treat primary type I osteoporosis.