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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。
BACKGROUND: Synthetic osteogenic growth peptide stimulates bone marrow mesenchymal stem cell proliferation and differentiation into osteoblasts in vitro. It can increase bone density and improve osteoporosis in vivo. However, the specific mechanism of action is not yet clear. OBJECTIVE: To screen differentially expressed genes of OPG - / - mouse bone marrow mesenchymal stem cells (BMSCs) under the action of osteogenic growth peptide by mouse whole genome microarray and to explore the possible gene and signal transduction pathways affected by osteogenic growth peptide. Methods: The whole genome Oligo chip provided by Shanghai Bohao Biotechnology Co., Ltd. was used to screen differentially expressed genes of OPG - / - mouse bone marrow mesenchymal stem cells (BMSCs) in osteoblast growth inhibitory group and blank control group. Real-time quantitative PCR analysis verified Some differentially expressed genes related to proliferation and differentiation, and combined with cluster analysis and path analysis to explore the mechanism of osteogenic growth peptide. RESULTS AND CONCLUSION: The results of the chip showed that 346 genes were down-regulated and 121 genes were up-regulated after osteogenic growth peptide effect. PCR validation results and chip results. The BioCarta pathway analysis involved six pathway proteins; the KEGG pathway analysis involved 12 pathways. The effect of osteogenic growth peptide on OPG - / - mice bone marrow mesenchymal stem cells involves multiple signal transduction pathways, of which MAPK signaling pathway may play a crucial role in its proliferation.