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

目的从力学生物学角度研究渗透压对未成熟树突状细胞(immature dendritic cells,im DCs)生物力学特性和免疫功能的影响。方法不同渗透压处理im DCs后,CCK-8检测细胞存活率,激光共聚焦显微镜观察细胞骨架结构的变化,细胞电泳仪检测细胞电泳迁移率的改变,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,流式细胞仪检测细胞的抗原吞噬能力。结果高、低渗均会改变im DCs的F-actin结构,甚至诱导细胞凋亡。低渗组电泳率显著高于等渗组,高渗组电泳率低于等渗组(P<0.05)。荧光偏振结果显示,高、低渗均会显著降低细胞的膜流动性(P<0.05);q PCR结果发现,高、低渗会显著上调im DCs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im DCs生物力学特性和免疫表型分子表达,研究结果对深入理解DCs的免疫调节功能具有重要意义。

Effects of Osmotic Pressure on Biomechanical Properties and Immune Functions of Immature Dendritic Cells

Objective To explore the effects of osmotic pressure on biomechanical properties and immune function of immature dendritic cells (imDCs) from mechanobiological viewpoint. Methods After treated with different osmotic pressures, the cell viability of imDCs was detected using cell counting kit-8 (CCK-8). The changes in morphology of imDCs were observed under 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). The phagocytic ability of the cell was detected by flow cytometry. ResultsBoth hyperosmosis and hypoosmosis could remodel the cyoskeletonof cells, even induce apoptosis. The electrophoresis mobility of the hypoosmosis group was significantly higher than that of the normal osmolarity group, while that of the hyperosmosis group was lower than that of the normal osmolarity group (P<0-05). Fluorescence polarization results showed that both hyperosmosis and hypoosmosis could significantly decrease the membrane fluidity of cells (P<0-05). The results 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, and the phagocytosis of cell was increased (P<0-05). Conclusions Hypertonic and hypotonic stress can influence biomechanical properties of imDCs and expression of immune-related molecules. The research findings are important for further understanding the immune regulation function of DCs.