骨形态发生蛋白2诱导C2C12和MC3T3-E1的成骨分化与自噬

发布时间：2018-01-09 00:14

本文关键词：骨形态发生蛋白2诱导C2C12和MC3T3-E1的成骨分化与自噬　出处：《中国组织工程研究》2014年20期 　论文类型：期刊论文

【摘要】：背景:一系列研究表明自噬与分化有密切联系;骨形态发生蛋白2是诱导C2C12、MC3T3-E1成骨分化经典途径,是研究成骨分化过程的理想模型。目的:观察自噬与骨形态发生蛋白2诱导细胞株C2C12、MC3T3-E1成骨分化的关系。方法:Real-Time PCR检测MC3T3-E1与C2C12在骨形态发生蛋白2(100μg/L)诱导培养3 d后成骨与自噬相关指标变化。碱性磷酸酶染色检测不同浓度3-甲基腺嘌呤(0,1,5,10 mmol/L)对骨形态发生蛋白2(100μg/L)诱导培养7 d MC3T3-E1与C2C12成骨指标碱性磷酸酶变化,Western Blot检测C2C12和MC3T3-E1在骨形态发生蛋白2(100μg/L)诱导不同时间点(0,12,24,48,72,96 h)LC3-Ⅰ/Ⅱ蛋白表达水平。结果与结论:在骨形态发生蛋白2诱导细胞株C2C12、MC3T3-E1成骨分化过程中,诱导自噬相关mRNA与蛋白水平均有增高趋势,且自噬相关蛋白LC3水平增高与时间相关。同时,抑制自噬成骨分化过程中碱性磷酸酶表达水平降低。因此,自噬与骨形态发生蛋白2诱导细胞株C2C12、MC3T3-E1成骨分化过程有密切关系。
[Abstract]:Background: a series of studies indicate that autophagy is closely linked with differentiation; bone morphogenetic protein 2 C2C12 induced osteogenic differentiation of MC3T3-E1 classical pathway, is the ideal model to study osteogenic differentiation. Objective: To observe the autophagy and bone morphogenetic protein 2 induced cell lines C2C12, MC3T3-E1 into bone differentiation. Methods: Real-Time PCR detection of MC3T3-E1 and C2C12 in bone morphogenetic protein 2 (100 g/L) after 3 D culture into the changes of bone and autophagy induced by alkaline phosphatase staining. Different concentrations of 3- methyl adenine (0,1,5,10 mmol/L) for the detection of bone morphogenetic protein 2 (100 g/L) 7 d MC3T3-E1 and C2C12 induced bone alkaline index Western Blot phosphatase changes, the detection of C2C12 and MC3T3-E1 in bone morphogenetic protein 2 (100 g/L) induced by different time points (0,12,24,48,72,96 h) the expression level of LC3- I / II protein. Results and conclusion: the occurrence of eggs in bone formation 2 white induced cell line C2C12, MC3T3-E1 in the process of osteogenic differentiation, induction of autophagy related mRNA and protein level were significantly increased, and autophagy related protein LC3 levels increased with time. At the same time, inhibition of autophagy level of alkaline phosphatase decreased expression of bone differentiation. Therefore, autophagy and bone morphogenetic protein 2 induced cell line C2C12 MC3T3-E1, there is a close relationship between the osteogenic differentiation process.

【作者单位】：同济大学附属第十人民医院中心实验室;
【基金】：国家自然科学基金项目(81171178)~~
【分类号】：R363
【正文快照】： 0引言Introduction细胞自噬是一个动态过程,涉及细胞器与蛋白质向溶酶体或液泡运输过程中亚细胞膜的重排,细胞器与蛋白质在溶酶体和液泡中降解或循环[1-4]。自噬既广泛存在于正常的生理过程中,如清除细胞废物、结构重建、生长分化等,同时还参与多种疾病的病理过程。自噬不仅与

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