天然羟基磷灰石对间充质干细胞作用的分子机理初步研究
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天然羟基磷灰石对间充质干细胞作用的分子机理初步研究
张志伟,王健丹,吕晓迎
基金项目:中国教育部博士点基金 (No.20100092110027);973计划项目(No. 2009CB930000);国家自然科学基金 (No. 31170910, 31271012, 30770583);江苏省研究生培养创新工程基金(CX10B_082Z) 作者简介:张志伟(1981-),男,博士研究生,主要研究方向:生物材料,生物相容性 通信联系人:吕晓迎(1956-),女,教授,主要研究方向:生物材料,生物相容性,生物电子学.

(东南大学生物电子学国家重点实验室,南京,210096) 5
摘要:本论文的目的是通过对细胞生物学和生物组学的综合研究,探讨天然羟基磷灰石(NHA)对骨髓间充质干细胞(MSCs)的作用机理;首先从猪骨中提取NHA,并制备成圆盘状试样,然后采用细胞生物学方法研究MSCs在NHA圆盘状试样表面的增殖情况;接着采用蛋白质组学技术研究培养于NHA表面的MSCs细胞的蛋白表达谱情况,接着进行GO分类分析并与前期10 基因芯片数据的GO分类分析进行比较,最后对蛋白质组学和基因芯片数据进行通路的联合分析。结果显示NHA表面培养的MSCs细胞增殖率下降;培养于NHA表面的MSCs细胞的蛋白表达谱所涉及GO功能广泛,且多数GO功能与基因芯片的GO功能相一致;差异表达的基因/蛋白参与了89条通路,其中MAPK通路可能在NHA诱导MSCs成骨分化中发挥重要作用。总之,NHA对MSCs的作用涉及基因转录和蛋白质表达各层次,NHA可能通过对MAPK通路的影15 响,最终诱导了MSCs细胞的成骨分化。
关键词:生物材料;天然羟基磷灰石;转录组学;蛋白质组学;生物学通路
中图分类号:R318.08
Pilot Study of molecular mechanism underlying 20 MSCs–natural hydroxyapatite interaction
ZHANG Zhiwei, WANG Jiandan, Lü Xiaoying
(State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096)
Abstract: This work combined cytobiology and biomics analysis to elucidate the mechanism of the interaction between natural hydroxyapatite (NHA) and mesenchymal stem cells (MSCs). 25 Firstly, NHA power obtained from pig bones was fabricated into disc-shaped sample. Subsequently, the proliferation of MSCs cultured on the disc-shaped NHA was investigated. Then, The proteomics approach was employed to detect protein expression profiling of MSCs cultured on NHA. The biologically relevant gene ontology (GO) terms of differentially expressed proteins was analyzed and compared with GO result of previous transcriptomics data. Finally, an 30 integrating pathway study of proteomics and transcriptomics data was analyzed. The results showed that the cell proliferation rate was decreased on NHA. Protein expression profiling of MSCs cultured on NHA were involved in several GO functional classification. Comparing the GO analysis results from proteomics and gene expression peofile, we found that the differentially expressed genes and proteins shared many GO terms. The genes and proteins involved in 89 35 pathways were shown to be differentially expressed. Among them, MAPK pathways might played an important role in NHA-induced osteogenic differentiation. In conclusion, NHA affected MSCs via both transcription and translation level. NHA-induced osteogenic differentiation of MSCs eventually occurred through the MAPK pathway.
Key words: Biomaterial, Natural hydroxyapatite, Transcriptomics, Proteomics, Biological 40 pathways