Experimental study on the regulation of phagocytic function and polarization of macrophages by exosomes from patients with endometriosis
Objective To explore the effects of exosomes derived from endometriosis on phagocytic function and polarization of macrophages.Methods Exosomes were isolated from the stromal cells of healthy individuals,named the con-trol(CON)group,and from the stromal cells of patients with endometriosis,named the endometriosis(EMS)group.Nanopar-ticle tracking technology was used to detect the size,particle size distribution,and potential of exosomes in each group.West-ern blot was performed to analyze the expression of exosome markers CD9 and CD63 in the CON and EMS groups.The exosomes from the CON and EMS groups were co-cultured with THP-1 macrophages and named the CON-exosomes group and EMS-exosomes group,respectively.The PBS-Control group was used as a blank control and treated with an equal amount of PBS.MTT assay was used to evaluate the proliferation activity of the three groups of cells.Flow cytometry was performed to assess the apoptosis rate,as well as the ratio of M1-polarized(CD86+)and M2-polarized(CD206+)macrophages in each group.Fluorescent dye CM-Dil was used to label exosomes,DAPI was used to stain cell nuclei,and confocal microscopy was used to observe the phagocytic ability of macrophages towards exosomes.Results There was no significant difference in the physical parameters of exosomes between the CON and EMS groups(both P＞0.05),with diameters ranging from 95.67 to 119.25 nm and 94.82 to 121.33 nm,and peak size of particle size distribution at 103.46 nm and 106.89 nm,respectively.The Zeta potentials of exosomes in the CON and EMS groups were(-29.28±3.91)mV and(-28.64±4.52)mV,respectively(P＞0.05).There was no significant difference in the expression of exosome markers CD9 and CD63 between the two groups(P＞0.05).Compared to the PBS-Control group,the THP-1 cell proliferation activity and M2 polarization ratio decreased in the CON-exosomes group(P＜0.05),while the apoptosis rate showed no significant difference(P＞0.05).Compared to the CON-exosomes group,the cell proliferation activity and M2 polarization ratio increased,and the apoptosis rate increased in the EMS-exosomes group(P＜0.05).Compared to the CON-exosomes group,the THP-1 cell phagocytic function towards exosomes was weakened in the EMS-exosomes group.Conclusion Exosomes derived from endometriosis inhibit the prolif-eration activity and M2 polarization of THP-1 cells,promote apoptosis,and weaken their phagocytic function.
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