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目的观察锰暴露诱导BV2小胶质细胞活化过程中COX2分子表达的变化。方法实验中BV2细胞分3组,为对照组,50μM锰暴露组和100μM锰暴露组,分别处理12、24和48 h。免疫荧光化学染色方法观察BV2细胞锰暴露24h后的活化情况,荧光实时定量PCR(qRT-PCR)和Western Blot法分别检测COX2分子mRNA和蛋白表达水平的改变。结果免疫荧光化学染色结果显示,锰暴露24 h后BV2小胶质细胞形态发生改变,细胞胞体变大变圆,突起变短或消失,活化细胞占总细胞数的比例增加(P<0.05);实时定量PCR结果显示,锰暴露12 h和24 h后COX2 mRNA水平增加(P<0.05);Western Blot结果显示,锰暴露24 h和48 h后COX2蛋白表达水平增加(P<0.05)。结论锰暴露能够诱导BV2细胞COX2分子表达增加,COX2可能参与了锰诱导小胶质细胞活化的过程。
Objective To observe the changes of COX2 expression in activated BV2 microglial cells induced by manganese exposure. Methods BV2 cells were divided into 3 groups: control group, 50μM manganese exposure group and 100μM manganese exposure group for 12, 24 and 48 h respectively. Immunofluorescence staining was used to observe the activation of BV2 cells exposed to manganese for 24 hours. The expression of COX2 mRNA and protein were detected by real-time quantitative PCR (qRT-PCR) and Western Blot. Results The results of immunofluorescence staining showed that the morphology of BV2 microglial cells changed after 24 h of Mn exposure. The cell bodies became bigger and round, and the protrusions became shorter or disappeared. The ratio of activated cells to total cells increased (P <0.05). The results of real-time quantitative PCR showed that the COX2 mRNA levels increased after 12 and 24 h of manganese exposure (P <0.05). Western Blot showed that the expression of COX2 protein increased 24 and 48 h after Mn exposure (P <0.05). Conclusion Manganese exposure can induce the expression of COX2 in BV2 cells. COX2 may be involved in the process of manganese-induced microglial activation.