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Effects of lactoferrin on the differentiation of pluripotent mesenchymal cells.

机译:乳铁蛋白对多能间充质细胞分化的影响。

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Lactoferrin accelerates bone formation, but the precise cellular mechanism behind this is still unclear. We examined the effect of lactoferrin on the differentiation of pluripotent mesenchymal cells using a typical pluripotent mesenchymal cell line, C2C12. Cells were cultured in low-mitogen differentiation medium to induce cell differentiation, with or without the addition of lactoferrin. The cell lineage was determined by alkaline phosphatase (ALPase) activity, mRNA expression of cellular phenotype-specific markers using real-time polymerase chain reaction (PCR), and protein synthesis using Western blotting. The expression of low-density lipoprotein lipase receptor-related proteins (LRPs) 1 and 2, both lactoferrin receptors, was determined by reverse transcription-PCR. ALPase activity increased after the addition of lactoferrin. The mRNA expression of Runx2, osteocalcin, and Sox9 increased markedly as a result of lactoferrin treatment, whereas the expression of MyoD, desmin, and PPARgamma decreased significantly. Western blots showed that lactoferrin stimulation increased Runx2 and Sox9 proteins, whereas it decreased MyoD and PPARgamma synthesis. C2C12 cells expressed the LRP1 lactoferrin receptor. These results indicate that lactoferrin treatment converts the differentiation pathway of C2C12 cells into the osteoblastic and chondroblastic lineage.
机译:乳铁蛋白可加速骨骼形成,但其背后的确切细胞机制仍不清楚。我们使用典型的多能间充质细胞系C2C12检查了乳铁蛋白对多能间充质细胞分化的影响。在有或没有添加乳铁蛋白的情况下,在低促细胞分裂剂分化培养基中培养细胞以诱导细胞分化。通过碱性磷酸酶(ALPase)活性,使用实时聚合酶链反应(PCR)的细胞表型特异性标记物的mRNA表达以及使用蛋白质印迹的蛋白质合成来确定细胞谱系。通过逆转录PCR确定低密度脂蛋白脂酶受体相关蛋白(LRPs)1和2(均为乳铁蛋白受体)的表达。添加乳铁蛋白后,ALPase活性增加。乳铁蛋白处理后Runx2,骨钙素和Sox9的mRNA表达显着增加,而MyoD,结蛋白和PPARgamma的表达显着降低。蛋白质印迹显示乳铁蛋白刺激增加Runx2和Sox9蛋白,而降低MyoD和PPARgamma合成。 C2C12细胞表达LRP1乳铁蛋白受体。这些结果表明乳铁蛋白治疗将C2C12细胞的分化途径转化为成骨细胞和成软骨细胞谱系。

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