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目的通过Shotgun蛋白质组学分析,初步检测骨髓基质细胞(BMSCs)条件液中可能含有的蛋白质。方法将BMSCs条件液混匀后经超滤浓缩分为大于5kD和小于5kD两部分,并培养神经干细胞(NSCs),观察其后代中神经元、星形胶质细胞和少突胶质细胞的比例,鉴定出能调节NSCs分化的部分,并进行Shotgun蛋白质组学分析。结果BMSCs条件液分子量大于5kD的部分可以调节NSCs向神经元和少突胶质细胞方向分化,这部分条件液先经过聚丙烯酰胺凝胶电泳(SDS-PAGE)发现大部分蛋白集中在14kD以上,将蛋白条带酶解以后,经Shotgun分析共鉴定到456种蛋白质,其中154个相似蛋白质,17个假想蛋白质,56个未知蛋白质,剩余的229个蛋白质中大多为细胞骨架蛋白、分泌蛋白、信号转导蛋白、酶类和转运蛋白等。结论BMSCs分泌的多种蛋白质对NSCs的分化起调节作用,明确了BMSCs条件液中可能含有的成分。
Objective By Shotgun proteomics analysis, preliminary detection of bone marrow stromal cells (BMSCs) may contain the conditions of the protein. Methods BMSCs conditioned medium was mixed and concentrated by ultrafiltration into two groups of more than 5kD and less than 5kD, and the neural stem cells (NSCs) were cultured to observe the ratio of neurons, astrocytes and oligodendrocytes , Identified as part of the regulation of NSCs differentiation, and Shotgun proteomics analysis. Results The fraction of BMSCs with a molecular weight of more than 5 kD could regulate the differentiation of NSCs into neurons and oligodendrocytes. Most of the proteins were concentrated at 14 kD or more by SDS-PAGE. After proteolytic cleavage of protein bands, 456 proteins were identified by Shotgun analysis, including 154 similar proteins, 17 hypothetical proteins and 56 unknown proteins. Most of the remaining 229 proteins were cytoskeletal proteins, secretory proteins, signals Transducible proteins, enzymes and transporters. Conclusion A variety of proteins secreted by BMSCs play a regulatory role in the differentiation of NSCs, and the possible components of BMSCs conditioned solution are determined.