毛蕊异黄酮对氧糖剥夺再灌注BV2 小胶质细胞极化的影响

目的 探讨毛蕊异黄酮对氧糖剥夺再灌注BV2小胶质细胞极化的影响。方法 将对数生长期BV2细胞随机分为6组：正常组、模型组、尼莫地平组(5μg·mL^-1)、毛蕊异黄酮高剂量组(20μg·mL^-1)、毛蕊异黄酮中剂量组(10μg·mL^-1)、毛蕊异黄酮低剂量组(5μg·mL^-1)。氧糖剥夺3 h后，各给药组换成含药的完全培养基，复氧6 h。采用CCK-8法检测细胞活力；测定细胞上清液中的一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-10含量；免疫荧光双染法检测BV2细胞极化；WesternBlot法检测BV2细胞的iNOS、CD206蛋白表达水平。结果 与正常组比较，模型组的BV2细胞活力显著降低(P<0.01),NO、TNF-α、IL-1β水平明显升高(P<0.05),IL-10水平明显降低(P<0.05)；免疫荧光双染检测中M1型小胶质细胞标记物iNOS表达显著增强(P<0.01);Western Blot检测中iNOS蛋白表达明显上调(...

Effects of Calycosin on Microglia Polarization in BV2 Cells After Oxygen-Glucose Deprivation andReperfusion

To research the effect of calycosin on the polarization of BV2 microglia cells after oxygenglucosedeprivation and reperfusion. Methods The BV2 cells in logarithmic growth phase were randomly divided into6 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-glucosedeprivation for 3 hours， the drug-containing complete medium was replaced in each administration group， andreoxygenation 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 Western Blot. Results Compared with the normal group，the activity of BV2 cells in the model groupwas 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 M1microglia，was significantly increased in immunofluorescence double staining （P＜0.01） . The protein expression ofiNOS was significantly up-regulated in the detection of Western Blot （P＜0.05），and the protein expression of M2cell marker CD206 was significantly down-regulated in the detection of Wenstern Blot （P＜0.05） . Compared withthe model group，the viability of BV2 cells in the low-，medium- and high- dose calycosin groups and Nimodipinegroup 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 BV2cells 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 wassignificantly 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 itspolarization to M1 type，reduce the production of inflammatory mediators and oxidative damage，and protect braintissue damage after ischemia.