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目的:探讨七氟烷对培养的小鼠小胶质细胞中炎症因子表达的影响。方法:取新生(2~3天)C57BL/6小鼠,分离小胶质细胞,将其随机分为4组(n=10):对照组(Control);七氟烷组(Sevoflurane);NF-κB抑制剂组(PDTC);NF-κB抑制剂+七氟烷组(PDTC+Sevoflurane)。用Drager麻醉机向Sevoflurane组PDTC+Sevoflurane组培养的小胶质细胞盒内释放21%O2,5%CO2,4.1%七氟烷的气体,用气体分析仪持续监测各组的浓度。应用Iba-1的免疫荧光染色法对小鼠小胶质细胞进行纯度鉴定。分别在于给七氟烷后2 h、4 h和6 h时采用免疫印迹分析技术检测两组小胶质细胞IL-6和TNF-α的表达水平和NF-κB的活性。PDTC+Sevoflurane组在给七氟烷前一小时给予PDTC,采用ELISA技术和免疫印迹分析技术检测各组小胶质细胞IL-6和TNF-α的浓度和NF-κB的表达。结果:免疫印迹显示七氟烷组细胞中IL-6、TNF-α水平和NF-κB的激活水平升高;PDTC降低了七氟烷作用后核内NF-κB的表达,减弱了IL-6和TNF-α水平的升高作用。结论:七氟烷可通过激活NF-κB信号通路,进一步激活培养的小鼠小胶质细胞中炎症因子的表达。
Objective: To investigate the effect of sevoflurane on the expression of inflammatory cytokines in cultured mouse microglia. Methods: Newborn (2 ~ 3 days) C57BL / 6 mice were isolated and microglia were isolated and randomly divided into 4 groups (n = 10): Control group, Sevoflurane group, NF -κB inhibitor group (PDTC); NF-κB inhibitor + sevoflurane group (PDTC + Sevoflurane). Gasses of 21% O2, 5% CO2, 4.1% sevoflurane were released from the microglial cells cultured in the Sevoflurane group PDTC + Sevoflurane using a Drager anesthesia machine and the concentration of each group was continuously monitored with a gas analyzer. Mice microglia were identified by immunofluorescence staining with Iba-1. The expression of IL-6 and TNF-α and the activity of NF-κB in microglia were detected by Western blot analysis at 2 h, 4 h and 6 h after sevoflurane treatment respectively. PDTC + Sevoflurane group was given PDTC one hour prior to sevoflurane. The concentration of IL-6 and TNF-α and the expression of NF-κB in microglia were detected by ELISA and Western blot analysis. Results: Western blotting showed that the levels of IL-6, TNF-α and NF-κB were increased in sevoflurane group. PDTC decreased the expression of NF-κB in the nucleus and decreased the expression of IL-6 And TNF-α levels increased role. Conclusion: Sevoflurane can activate the NF-κB signaling pathway to further activate the expression of inflammatory cytokines in cultured mouse microglia cells.