渗透压对未成熟树突状细胞生物力学特性和免疫学功能的影响
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1. 贵州医科大学 基础医学院,贵阳;2. 贵州医科大学 生物与工程学院,贵州省免疫细胞与抗体工程研究中心/生物与医学工程重点实验室 贵阳;3.贵州医科大学 生物与工程学院,贵州省免疫细胞与抗体工程研究中心/生物与医学工程重点实验室 贵阳;4.贵州医科大学 基础医学院,贵阳;5.1. 贵州医科大学 基础医学院

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国家自然科学基金(11762006, 31771014, 31660258);贵州省科技计划项目(黔科合基础[2018]1412, 黔科合平台人才[2016]5676, 黔科合人才团队[2015]4021, 黔科合支撑[2019]2787号);2011协同创新中心(黔教合协同创新字[2015]04);贵州省细胞与基因工程创新群体(黔教合KY字[2016]031)


Effects of Osmotic Pressure on the Biomechanical Properties and Immune Functions of Immature Dendritic Cells
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1. School of Basic Medical Sciences, Guizhou Medical University, Guiyang, China;2. Immune Cells and Antibody Engineering Research Center of Guizhou Province/ Key Laboratory of Biological and Medical Engineering, School of Biology and Engineering, Guizhou Medical University, Guiyang;3.School of Basic Medical Sciences, Guizhou Medical University, Guiyang

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    摘要:

    目的 从力学生物学的角度研究渗透压(Osmotic pressure)对未成熟树突状细胞(immature dendritic cells, imDCs)生物力学特性和免疫功能的影响。方法 不同渗透压处理imDCs后,CCK-8检测细胞存活率,倒置显微镜、激光共聚焦显微镜观察细胞形态和细胞骨架结构的变化,细胞电泳仪检测细胞电泳率EPM (Cell electrophoresis mobility)的改变,荧光偏振法检测细胞的膜流动性,实时荧光定量PCR检测免疫相关分子的表达变化,流式细胞仪检查细胞的抗原吞噬能力。结果 高渗、低渗均会改变细胞的形态,甚至诱导细胞凋亡。低渗组的电泳率显著高于等渗组,高渗则反之(P<0.05)。荧光偏振结果显示高渗、低渗均会显著降低细胞的膜流动性(P<0.05),qPCR检测发现,高渗、低渗会显著上调imDCs免疫表型分子CCR7、CD40、CD205、CD11a、CD11c的表达和抗原吞噬能力(P<0.05)。结论 高、低渗应激会影响imDCs的生物力学特性和免疫表型分子的表达,这对深入理解DCs的免疫调节功能具有重要意义。

    Abstract:

    Objective To explore the effects of osmotic pressure on the morphology, biophysical properties, and expression levels of immunophenotype molecules on immature dendritic cells (imDCs) from mechanobiological viewpoint. Methods After treated with different osmotic pressures, the survival rate of DCs was detected using Cell Counting Kit-8 (CCK-8). The changes in morphology and size of DCs were observed under a Laser scanning confocal microscope. Cell electrophoresis was applied to detect the changes in cell electrophoresis mobility, The membrane fluidity of the cells was detected by fluorescence polarization method, and the expression changes of immune-related molecules were detected by real-time fluorescent quantitative PCR (qPCR). Results Both hyperosmosis and hypoosmosis could change the shape and size of cells, even induce apoptosis. The electrophoresis rate of the low permeability group was significantly higher than that of the equal permeability group, while that of the hypoosmosis group was lower (P<0.05). Fluorescence polarization results showed that both hyperosmosis and hypoosmosis could significantly decrease the membrane fluidity of cells (P<0.05). The result of qPCR detection showed that both hyperosmosis and hypoosmosis could significantly increase the expression of CCR7, CD40 ,CD205, CD11a, CD11c on the surface of DCs (P<0.05). Conclusion Hypertonic and hypotonic stress can change the electrophoresis rate, membrane fluidity, and the expression of immune-related molecules, which is important for understanding the immune regulation function of DCs.

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  • 收稿日期:2019-03-12
  • 最后修改日期:2019-04-29
  • 录用日期:2019-04-30
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