Abstract: ［Abstract］ Objective To investigate the regulation of isoflurane on microglial NLR family pyrin domain containing 3（NLRP3）inflammasome?related inflammatory microenvironment. Methods Microglia cell line（BV2）was pretreated or not with lipopolysaccharides（LPS）for 30 minutes, subsequently exposed to 4% isoflurane for 6 hours. The mRNA of NLRP3 was detected by quantitative reverse transcription?polymerase chain reaction（qRT?PCR）. The protein of NLRP3 and caspase?1 P20 was analyzed by Western?blot. Production of IL?1β in the supernatants was measured by enzyme linked immunosorbent assay（ELISA）. Toxicity of isoflurane were evaluated by CCK8 cell viability assay. Results Isoflurane activated downstream of NLRP3 pathway, increased the secretion of IL?1β. There was no significant impact on BV2 cell viability, when treated with isoflurane at the experimental concentration. Conclusion Isoflurane induced microglial inflammation through the NLRP3 inflammasome pathway. NLRP3 priming is necessary in Isoflurane induced IL?1β production.

Key words: NLRP3 inflammasome, isoflurane, neuroinflammation, postoperative cognitive dysfunction

摘要： ［摘要］目的 探讨异氟醚处理对小胶质细胞NLR家族热蛋白结构域包含蛋白3（NLR family pyrin domain containing 3，NLRP3）炎性小体通路相关炎症微环境的调控作用。方法 脂多糖（LPS；1 μg·mL?1）预活化小胶质细胞株（BV2）30 min，分别采用异氟醚单独处理和LPS预活化后异氟醚处理6小时。实时荧光定量聚合酶链反应（qRT?PCR）检测NLRP3的mRNA表达量；蛋白免疫印迹（Western?blot）分析NLPR3蛋白表达变化；酶联免疫吸附试验（ELISA）检测炎症因子白介素?1β（interleukin?1β，IL?1β）的分泌情况；CCK8细胞活性试验评估异氟醚处理的毒性作用。结果 qRT?PCR及Western?blot结果表明异氟醚单独处理不引起BV2细胞的NLRP3表达变化，LPS预活化后异氟醚处理上调预活化BV2细胞的NLRP3表达；ELISA结果表明异氟醚单独处理不引起BV2细胞IL?1β分泌，LPS预活化后异氟醚处理促进活BV2细胞IL?1β分泌；各组实验条件下BV2细胞活性均无显著改变。结论 异氟醚可通过激活NLRP3炎性小体调控小胶质细胞IL?1β的表达。

关键词: NLRP3 炎性小体, 神经炎症, 认知功能障碍, 异氟醚