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目的观察依达拉奉对离体培养大鼠小胶质细胞缺血再灌损伤后炎症因子表达的影响,以及Janus激酶2/信号传导子和转录激活子3(JAK2/STAT3)信号通路是否参与依达拉奉的神经保护。方法离体培养SpragueDawley乳鼠小胶质细胞,将细胞接种于细胞培养板随机分为3组(n_i=20):正常对照组(C组)常规培养;谷氨酸组(Glu组)给予含有1 mmol/L的Glu的培养基孵育30 min建立小胶质细胞缺血再灌损伤模型;依达拉奉组(Eda组)于细胞建立损伤模型后换成含有100μmol/L的Eda培养基孵育24 h;Glu损伤后24 h通过CCK-8测细胞存活率,Real-Time PCR检测JAK2、STAT3的mRNA水平;用ELASA法测定TNF-α和IL-2的变化;用流式细胞仪测细胞凋亡率。结果与C组比较,Glu组细胞存活率降低,JAK2和STAT3的mRNA表达增加,细胞上清液的TNF-α和IL-2浓度增加,细胞凋亡率增加(P<0.05);与Glu组比较,Eda组细胞存活率增加,JAK2和STAT3的mRNA表达减少,细胞上清液中TNF-α和IL-2浓度降低,细胞凋亡率下降(P<0.05)。结论依达拉奉减少了大鼠小胶质细胞缺血再灌注损伤后TNF-α和IL-2表达,JAK2/STAT3信号通路部分参与了炎症因子的表达。
Objective To investigate the effect of edaravone on the expression of inflammatory cytokines in cultured rat microglial cells after ischemia-reperfusion injury and the involvement of Janus kinase 2 / signal transducers and activator of transcription 3 (JAK2 / STAT3) signaling pathway Neuroprotection of edaravone. Methods The microglia of Sprague Dawley rats were cultured in vitro. The cells were inoculated into the cell culture plate and were randomly divided into 3 groups (n_i = 20): normal control group (C group) routine culture; glutamate group (Glu group) 1 mmol / L Glu for 30 min to establish the model of microglia ischemia-reperfusion injury. Edaravone group (Eda group) was used to establish the injury model and then incubated with Eda medium containing 100 μmol / L 24 h; Glu cell viability was measured by CCK-8 24 h after Glu injury; Real-time PCR was used to detect mRNA levels of JAK2 and STAT3; Changes of TNF-α and IL-2 were measured by ELASA; Cell viability was measured by flow cytometry Apoptosis rate. Results Compared with group C, the survival rate of Glu group was decreased, the mRNA expression of JAK2 and STAT3 was increased, the levels of TNF-α and IL-2 in supernatant were increased and the apoptosis rate was increased (P <0.05) Compared with Eda group, the cell viability increased, the mRNA expression of JAK2 and STAT3 decreased, the concentration of TNF-α and IL-2 decreased, and the apoptosis rate decreased (P <0.05). Conclusion Edaravone can reduce the expression of TNF-α and IL-2 in rat microglia after ischemia-reperfusion injury, and JAK2 / STAT3 signaling pathway is involved in the expression of inflammatory cytokines.