龟鹿二仙汤含药血清对大鼠成骨细胞增殖功能、OPG及ODF系统的研究  

Study of Guilu Erxian Decoctionon on Rats' Proliferation of Bone Cell,OPG and ODF System

作　　者：钱康[1] 

机构地区：[1]杭州市第一人民医院中医科,浙江杭州310006

出　　处：《中华中医药学刊》2014年第6期1478-1481,I0016,共5页Chinese Archives of Traditional Chinese Medicine

摘　　要：目的:探讨龟鹿二仙汤含药血清对大鼠成骨细胞(osteoblast,OB)增殖功能、OB关键基因骨保护素(OPG)、破骨细胞分化因子(ODF)的mRNA表达变化的研究,分析龟鹿二仙汤防治骨质疏松症的作用机理。方法:①采用胰蛋白酶-Ⅱ型胶原同浓度(10%、15%、20%)的龟鹿二仙汤含药血清培养大鼠成骨细胞(OB)24、48、72 h,应用MTT比色法来检测其对OB增殖功能的影响。②采用荧光定量PCR测定OB中OPG、ODF的表达。结果:龟鹿二仙汤含药血清对OB的增殖作用呈浓度依赖性,即20%>15%>10%;在培养24、48 h、72 h后显著促进其增殖作用。OPG mRNA表达在龟鹿二仙汤含药血清20%浓度时最强,且明显高于对照组(P<0.01),而ODF基因mRNA表达在20%浓度时最低,且明显低于对照组(P<0.01)。结论:龟鹿二仙汤含药血清对体外培养SD大鼠OB增殖具显著作用且呈浓度依赖性,可在分子水平上调节OPG、ODF表达,可能与其治疗骨质疏松机制相关。Objective ： To investigate the effect of serum containing Guilu Erxian Decoction on rat osteoblasts （ osteoblast, OB） proliferation, the expression of mRNA of osteoprotegerin （OPG） and osteoclast differentiation factor （ ODF）, and to discuss the mechanism of Guilu Erxian Decoction in the prevention and treatment of osteoporosis. Methods ： （1）By trypsintype II collagen with concentration （10%, 15% ,20% ） of rat osteoblasts （OB） were cultured in serum containing Guilu Erxian Decoction 24,48,72 h. MTT colorimetric method was used to detect the effect on proliferation of OB function. （2）The expressions of OPG and ODF were detected by fluorescence quantitative PCR assay. Results： The proliferation effect of serum containing Guilu Erxian Decoction on OB in a concentration dependent manner,i, e. 20% 〉 15% 〉 10%. It can significantly enhance the proliferation effect in training 24,48,72 h. OPG mRNA expression was the strongest in 20% serum containing Guilu Erxian Decoction, and significantly higher than that of the control group （ P 〈 0.01 ） , and the ODF gene mRNA expression was the lowest at 20% concentration, and significantly lower than that of the control group （ P 〈0.01 ）. Conclusion： The serum containing Guilu Erxian Decoction has significant effects on OB proliferation of SD rats in a dose - dependent manner in vitro, and can regulate OPG and ODF expressions at the molecular level, which may be related to the mechanism of treating osteoporosis.

关 键 词：龟鹿二仙汤 含药血清 成骨细胞 骨保护素 破骨细胞分化因子 

分 类 号：R285.5[医药卫生—中药学]