Effects of simulated microgravity on biological function of mouse macrophage RAW264.7

LI Meng; ZHANG Shu; HU Zebing; SUN Quan; XU Liqun; ZHANG Zhengxiang; SHI Fei

doi:10.13276/j.issn.2097-1656.2024.02.004

Abstract：Objective To explore the effects of simulated microgravity on the biological functions of mouse macrophages(RAW264.7)by using 2D clinostat to simulate the biological effects of microgravity,and to provide theoretical basis and data support for further research on the immunosuppressive mechanism of space microgravity.Methods RAW264.7 cells were divided into normal gravity(NG)group and simulated microgravity(SMG)group.After 72 h of simulated microgravity with 2D clinostat,the proliferation ability of RAW264.7 macrophages was detected by CCK-8 and EdU assays.The wound-healing assay and Transwell migration assay were performed to test the metastasis capacity of macrophages.The cells were labeled with DCFH-DA and the changes of reactive oxygen species(ROS)in macrophages were detected by inverted fluorescence microscopy.Results The proliferation of RAW264.7 macrophages was inhibited after simulated microgravity for 72 h.The results of CCK-8 assay showed that the cell proliferation rate of NG group was significantly higher than that of SMG group(P＜0.05).The results of EdU fluorescence staining showed that there were fewer positive cells in SMG group than those in NG group(P＜0.01).The wound-healing assay and Transwell assay showed that 72 h of simulated microgravity had a significant effect on the migration ability of macrophages.Compared with NG group,simulated microgravity could enhance the migration ability of RAW264.7 cells(P＜0.05).DCFH-DA labeling showed that simulated microgravity significantly inhibited ROS secretion of macrophage(P＜0.01).Conclusion Clinostat-based simulated microgravity has an effect on the biological functions of RAW264.7 cells,which is mainly manifested in the enhanced migration ability of RAW264.7 cells,and the inhibition of cell proliferation and ROS secretion ability.