FTY720-P对成骨细胞分化成熟的作用
发布时间：2018-11-03 12:27
【摘要】：目的以FTY720-P作用于成骨细胞后,分析相关蛋白和基因表达的变化,明确FTY720-P对成骨细胞的作用机制。方法取Wistar大鼠乳鼠颅盖骨进行成骨细胞原代培养。将不同浓度的FTY720-P(0、200、300、400 ng/m L)作用于成骨细胞,MTT试验和流式细胞术进行生长速率和细胞周期的检测,碱性磷酸酶(ALP)染色和钙化结节染色对比FTY720-P用药前后对细胞形态学功能的影响,并分别采用ELISA、Western blot和Real Time RT-PCR检测相关蛋白和基因的表达。结果 MTT显示FTY720-P对细胞生长影响不大。流式细胞术结果提示,细胞阻滞在G_2期,对周期影响不大。ALP染色显示对照组蓝紫色颗粒更加明显;茜素红染色可见数量较多和更加明显的钙化结节。PCR结果中ALP的表达是下调的,骨钙素(OCN)和胰岛素样生长因子(IGF)的表达是上调的;而在ELISA实验中,OCN和IGF的分泌是增加的,Western blot检测中,胞外胶原蛋白I型分泌也增加,而在ALP活性实验中,胞内ALP活性是降低的。结论 FTY720-P可以促进成骨细胞细胞外基质的成熟和矿化,但对细胞增殖和周期没影响。
[Abstract]:Objective to investigate the effect of FTY720-P on osteoblasts by analyzing the changes of protein and gene expression in osteoblasts. Methods the calvaria of Wistar rats was cultured in primary culture. Osteoblasts were treated with different concentrations of FTY720-P (0200300400 ng/m L). The growth rate and cell cycle were measured by MTT test and flow cytometry. Alkaline phosphatase (ALP) staining and calcified nodule staining were used to compare the effects of FTY720-P on cellular morphology, and ELISA,Western blot and Real Time RT-PCR were used to detect the expression of related proteins and genes. Results MTT showed that FTY720-P had little effect on cell growth. The results of flow cytometry showed that the cells were blocked in the second phase of Gal and had little effect on the cycle. ALP staining showed that the blue purple granules were more obvious in the control group. The expression of ALP was down-regulated and the expression of osteocalcin (OCN) and insulin-like growth factor (IGF) was up-regulated in PCR. In ELISA assay, the secretion of OCN and IGF was increased in, Western blot assay, and the type I secretion of extracellular collagen was also increased. In ALP activity test, the intracellular ALP activity was decreased. Conclusion FTY720-P can promote the maturation and mineralization of extracellular matrix of osteoblasts, but has no effect on cell proliferation and cell cycle.
【作者单位】： 广东医科大学研究生学院;广东省第二人民医院创伤骨科;南方医科大学研究生学院;
【基金】：广东省科技计划项目(编号:2016A020215124)
【分类号】：R68
[Abstract]:Objective to investigate the effect of FTY720-P on osteoblasts by analyzing the changes of protein and gene expression in osteoblasts. Methods the calvaria of Wistar rats was cultured in primary culture. Osteoblasts were treated with different concentrations of FTY720-P (0200300400 ng/m L). The growth rate and cell cycle were measured by MTT test and flow cytometry. Alkaline phosphatase (ALP) staining and calcified nodule staining were used to compare the effects of FTY720-P on cellular morphology, and ELISA,Western blot and Real Time RT-PCR were used to detect the expression of related proteins and genes. Results MTT showed that FTY720-P had little effect on cell growth. The results of flow cytometry showed that the cells were blocked in the second phase of Gal and had little effect on the cycle. ALP staining showed that the blue purple granules were more obvious in the control group. The expression of ALP was down-regulated and the expression of osteocalcin (OCN) and insulin-like growth factor (IGF) was up-regulated in PCR. In ELISA assay, the secretion of OCN and IGF was increased in, Western blot assay, and the type I secretion of extracellular collagen was also increased. In ALP activity test, the intracellular ALP activity was decreased. Conclusion FTY720-P can promote the maturation and mineralization of extracellular matrix of osteoblasts, but has no effect on cell proliferation and cell cycle.
【作者单位】： 广东医科大学研究生学院;广东省第二人民医院创伤骨科;南方医科大学研究生学院;
【基金】：广东省科技计划项目(编号:2016A020215124)
【分类号】：R68
【参考文献】
相关期刊论文 前7条
1 熊苗;朱洁萍;李莉;刘阳;季玉琴;王军;;过继转移FTY720-DC对自然流产模型孕鼠胚胎丢失率的影响[J];上海交通大学学报(医学版);2016年06期
2 王勇平;廖q，
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