【摘要】 目的:研究淫羊藿总黄酮(total flavonoids of herba ep im ed ii,HEF)及其大鼠含药血清(HEF-S)、去蛋白含药血清(NP-HEF-S)对体外培养鼠颅骨成骨细胞增殖及骨钙素(osteocalc in,OC)、骨桥蛋白(osteopontin,OP)和Ⅰ型胶原(typeⅠcollagen,COLⅠ)mRNA表达的影响,以了解淫羊藿治疗骨质疏松的可能物质基础,及其对成骨相关蛋白基因表达的作用。方法:HEF及其HEF-S、NP-HEF-S的冻干样品的稀释液分别与鼠颅骨成骨细胞共培养,MTT法测定其增殖率;将NP-HEF-S与鼠颅骨成骨细胞共培养,RT-PCR测定细胞OC、OP和COLⅠmRNA的表达。结果:HEF对细胞增殖无明显影响,且在终浓度为1mg/m l时产生抑制作用;HEF-S在各实验浓度(100~0.1倍原血药浓度)下均能显著促进成骨细胞增殖,且呈一定的剂量依赖关系;NP-HEF-S的作用相似但作用强度略低;10倍量的NP-HEF-S可使OC mRNA表达量明显提高,定量分析表明其表达量近5倍于空白对照组,但对OP和COLⅠmR-NA表达影响甚微。结论:淫羊藿治疗骨质疏松的直接物质基础可能并非HEF,而与其经口给予后吸收入血的HEF代谢产物及其诱生物有关。血清中的HEF代谢产物不仅可促进细胞的分化增殖,且促进其OC mRNA表达。更多还原

【Abstract】 Objective: To research the effects of total flavonoids of herba epimedii(HEF),rats serum after administration with HEF(HEF-S) and non-protein HEF-S(NP-HEF-S) on osteoblast proliferation of Clonal Murine Calvarial Osteoblast-like Cell Line(MC 3T3-E1) in vitro and the gene expression of osteocalcin(OC),osteopontin(OP),and type Ⅰcollagen(COLⅠ),so as to explore the possible effective substance of herba epimedii on treating osteoporosis and its effect on the gene expression of osteoblast related protein factors.Method: HEF-S and NP-HEF-S were prepared through effective substance and then were freeze dried.The dried serum would be diluted into different concentrations,and the concentration which is equal to that of incipient serum was defined 1 times concentration.HEF and both serum were cultivated with 3T3-E1 separately and the proliferation ratio was determined by MTT assay.The effect of NP-HEF-S on the gene expression of OC,OP and COL I were determined by RT-PCR.Results: HEF had no obvious effect on cell proliferation except the inhibition on 1mg/ml,whereas the HEF-S could enhance the osteoblast proliferation in dose dependent.NP-HEF-S was similar to HEF-S but behaved slightly weaker on the same concentration,and it made statistical discrepancy at the 0.1 times concentration compared with HEF-S.NP-HEF-S had no effect on OP and COL I mRNA expression,but it could increase the OC mRNA express amount about 5 times than that of control group at 10 times concentration.Conclusion: The effective substance of herba epimedii treating on osteoporosis is not HEF itself but certain of metabolites derived from HEF and the products induced by these metabolites in serum.The metabolite in serum could not only enhance the cell differentiation and proliferation,but also increase the expression of OC mRNA.更多还原