Effects of Calycosin on Microglia Polarization in BV2 Cells After Oxygen-Glucose Deprivation and Reperfusion
Objective To research the effect of calycosin on the polarization of BV2 microglia cells after oxygen-glucose deprivation and reperfusion.Methods The BV2 cells in logarithmic growth phase were randomly divided into 6 groups:normal group,model group,Nimodipine group(5 μg·mL-1),calycosin high-dose group(20 μg·mL-1),calycosin medium-dose group(10 μg·mL-1),calycosin low-dose group(5 μg·mL-1).After oxygen-glucose deprivation for 3 hours,the drug-containing complete medium was replaced in each administration group,and reoxygenation for 6 hours.CCK-8 was used to detect cell viability;the contents of NO,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-10 in the cell supernatant were measured.BV2 cell polarization was detected by double immunofluorescence staining.The protein expression levels of iNOS and CD206 in BV2 cells were detected by Western Blot.Results Compared with the normal group,the activity of BV2 cells in the model group was significantly decreased(P＜0.01),the levels of NO,TNF-α and IL-1β were significantly increased(P＜0.05),and the level of IL-10 was significantly decreased(P＜0.05).The expression of iNOS,a marker of M1 microglia,was significantly increased in immunofluorescence double staining(P＜0.01).The protein expression of iNOS was significantly up-regulated in the detection of Western Blot(P＜0.05),and the protein expression of M2 cell marker CD206 was significantly down-regulated in the detection of Wenstern Blot(P＜0.05).Compared with the model group,the viability of BV2 cells in the low-,medium-and high-dose calycosin groups and Nimodipine group was significantly increased(P＜0.01),the levels of NO,TNF-α and IL-1β were significantly decreased(P＜0.05),and the level of IL-10 was significantly increased(P＜0.05).The protein expression of iNOS in BV2 cells of calycosin(10 μg·mL-1)group was significantly decreased by immunofluorescence double staining(P＜0.01).Western Blot showed that the protein expression of iNOS in BV2 cells of calycosin(10 μg·mL-1)group was significantly down-regulated(P＜0.05),and the protein expression of CD206 was significantly up-regulated(P＜0.05).Conclusion Calycosin can promote the polarization of activated BV2 microglia to M2 type,inhibit its polarization to M1 type,reduce the production of inflammatory mediators and oxidative damage,and protect brain tissue damage after ischemia.
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