【摘要】 背景:合成成骨生长肽在体外可刺激骨髓间充质干细胞增殖和向成骨细胞分化,体内可增加骨密度改善骨质疏松,但具体作用机制尚未明确。目的:应用小鼠全基因组芯片筛查合成成骨生长肽作用下OPG-/-小鼠骨髓间充质干细胞的差异表达基因,探索成骨生长肽作用下可能影响到的基因与信号传导通路。方法:应用上海伯豪生物技术有限公司提供的小鼠全基因组Oligo芯片,筛选成骨生长肽干预组与空白对照组OPG-/-小鼠骨髓间充质干细胞差异表达基因,实时定量PCR分析验证部分与增殖分化相关的差异表达基因,并结合聚类分析及通路分析来探索成骨生长肽的作用机制。结果与结论:芯片结果显示,成骨生长肽作用后使346条基因表达下调,121条基因表达上调。PCR验证结果与芯片结果相符。经BioCarta通路分析,涉及6条通路蛋白;经KEGG通路分析,涉及12条通路。成骨生长肽对OPG-/-小鼠骨髓间充质干细胞的作用涉及多条信号传导通路,其中MAPK信号传导通路可能对其促增殖起着至关重要作用。更多还原

【Abstract】 BACKGROUND:Synthetic osteogenic growth peptide can stimulate proliferation and osteoblast differentiation of bone mesenchymal stem cells(BMSCs) in vitro,and which can enhance bone density in vivo,but the mechanism of the effect of osteogenic growth peptide(OGP) on BMSCs is not clear yet.OBJECTIVE:To investigate the effect of OGP on gene expression changes in BMSCs using mouse whole-genome microarray and to explore the gene and signal transduction pathway influenced by OGP.METHODS:Mouse whole-genome Oligo chip was obtained from Shanghai Bo Hao Biotechnology Co.,Ltd.,gene expression changes in BMSCs of OPG-/-in the OGP group(treated with OGP) and control group were screened.Real-time quantitative reverse transcriptase-polymerase chain reaction(RT-PCR) was used to analyze and verify gene expression changes related to proliferation and differentiation,and which combined with cluster analysis and pathway analysis to explore the mechanism of OPG.RESULTS AND CONCLUSION:Chip results showed that 346 genes expression down-regulated by OPG,121 genes expression up-regulated.PCR validation results were consistent with the chip.Six pathways proteins were identified by BioCarta pathway analysis and 12 pathways were identified by KEGG pathway analysis.Multiple pathways were related to the mechanism of the effect of OGP on gene expression changes in BMSCs of OPG-/-mice,among which the MAPK signal transduction pathway may play an important role in their proliferation.更多还原