Stomatological hospital of Chongqing medical university, Chongqing, P. R. China.
Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, Chongqing, P. R. China.
Proteomics. 2018 Feb;18(3-4). doi: 10.1002/pmic.201700215. Epub 2018 Feb 2.
Hypoxic preconditioning is commonly applied to enhance mesenchymal stem cells (MSCs) therapeutic effect before transplantation. Elucidating the effect of hypoxic preconditioning would be beneficial for improved application. However, the influence of hypoxia on dental tissue derived MSCs cultured in 3D was unknown. Thus, the present study is to investigate gene expression and proteome of dental pulp stem cells (DPSCs) after hypoxic preconditioning. DPSCs were isolated, cultured in a 3D system under the normoxic and hypoxic conditions. The gene expression was examined with reverse transcription polymerase chain reaction, and the proteome was analyzed using iTRAQ-based mass spectrometry. The expressions of HIF-1α, VEGFA, KDR at mRNA level was upregulated while BMP-2 was downregulated. Two thousand one hundred and fifteen proteins were identified and 57 proteins exhibited significant differences after hypoxic preconditioning (30 up-regulated, 27 down-regulated). Bioinformatic analysis revealed the majority of up-regulated proteins are involved in cellular process, angiogenesis, protein binding and transport, regulation of response to stimulus, metabolic processes, and immune response. Increased IL-6 and decreased TGF-1β protein expression under hypoxic condition were verified by ELISA. Hypoxic preconditioning partly affected the gene and protein expression in DPSCs under 3D culture and may enhance the efficacy of MSCs transplantation.
缺氧预处理常用于增强间充质干细胞（MSCs）在移植前的治疗效果。阐明缺氧预处理的影响将有助于改善其应用。然而，缺氧对三维培养的牙源性组织来源的间充质干细胞的影响尚不清楚。因此，本研究旨在探讨缺氧预处理后牙髓干细胞（DPSCs）的基因表达和蛋白质组学变化。分离 DPSCs，在常氧和缺氧条件下进行三维培养。采用逆转录聚合酶链反应检测基因表达，采用 iTRAQ 基于质谱分析蛋白质组学。结果显示，缺氧预处理后 HIF-1α、VEGFA、KDR 在 mRNA 水平上调，BMP-2 下调。鉴定出 2115 种蛋白质，其中 57 种蛋白质在缺氧预处理后差异表达（30 种上调，27 种下调）。生物信息学分析表明，上调蛋白主要参与细胞过程、血管生成、蛋白结合和转运、对刺激的反应调节、代谢过程和免疫反应。ELISA 验证了缺氧条件下 IL-6 蛋白表达增加，TGF-β1 蛋白表达减少。缺氧预处理在三维培养条件下部分影响 DPSCs 的基因和蛋白质表达，可能增强 MSCs 移植的疗效。
