水菠菜叶乙醇提取物对MG-63骨质疏松相关基因表达的影响

摘要

为探讨水菠菜叶乙醇提取物与抗骨质疏松作用的关系及其作用机制,用水菠菜叶乙醇提取物处理人成骨样细胞MG-63 48 h后,提取水菠菜叶乙醇提取处理组和对照组细胞的总RNA,将两组RNA纯化为mRNA,逆转录成cDNA,用Cy3和Cy5两种不同的荧光染料进行线性扩增标记,然后与骨质疏松相关基因表达谱芯片杂交,扫描后对获得的数据用LuxScan 3.0软件分析.研究发现经水菠菜叶乙醇提取物诱导后MG-63细胞的244个骨质疏松相关基因中,有c-Fo、JNK、ODF、c-src、OSCAR、RANKL、Samd4、Samd6、Samd7 9个基因表达明显上调,LRP5、Dkk1、TGFB、OPG 4个基因明显下调.结果表明水菠菜叶乙醇提取物能改变骨质疏松相关基因的表达,抗骨质疏松作用机制可能与Wnt/β-catenin蛋白信号传导途径和骨保护素/核因子受体激活信号传导途径有关,与雌激素内分泌途径无关.%To investigate the effects of methanol extract from Rumex crispus leaf on osteoporosis-associated gene expression in MG-63 cells by gene expression profile chip.The total RNAs were extracted from methanol extract from Rumex crispus leaf untreated or treated on MG-63 cells to synthesize double-stranded cDNAs by reverse transcription.Then cDNA labeled with the incorporation o f Cy3 and Cy5-dUT were amplified linearly.Probes were hybridized with oligo-nucleotide microarray.The chips were scanned and afterwards analyzed by LuxScan software.After methanol extract from Rumex crispus leaf induction,expressions of 9 out of 244 osteoporosis related genes,including c-Fos,JNK,ODF,c-src,OSCAR,RANKL,Samd4,Samd6 and Samd7,were down-regulated dramatically.Meanwhile,expressions o f 4 out of 244 cancer genes,including LRP5,Dkk 1,TGFB and OPG,were up-regulated significantly.It could be concluded that methanol extract from Rumex crispus leaf would change the relative gene expression.The anti-osteoporosis mechanism may be related to the Wnt/β-catenin protein signaling pathway and OPG/nuclear factor receptor activation signal pathway.However,it was not involved in estrogen endocrine pathway.