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目的:探讨雷帕霉素对1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinium iodide，MPP＋)诱导的小胶质细胞中Nod样受体蛋白3(nod-like receptor protein 3，NLRP3)炎症小体激活的影响。方法:将BV2小胶质细胞分为对照组、模型组和雷帕霉素组，模型组和雷帕霉素组以MPP＋激活NLRP3炎症小体，雷帕霉素组予雷帕霉素预处理。实时荧光定量PCR(quantitative real-time PCR，RT-qPCR)检测NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein，ASC)和caspase-1的mRNA水平，免疫荧光检测NLRP3和白细胞介素-1β(interleukin-1β，IL-1β)的表达情况，Western blot检测NLRP3、ASC、caspase-1、beclin1和微管相关蛋白l轻链3(microtubule-associated protein 1 light chain 3，LC3)的表达情况。结果:模型组的NLRP3、ASC和caspase-1 mRNA水平较对照组升高(n t＝4.825，3.015，5.853，均n P<0.05)，雷帕霉素组的NLRP3和caspase-1 mRNA水平较模型组降低(n t＝2.75，2.89，均n P<0.05)。模型组NLRP3(1.54±0.22)、ASC(1.02±0.13)和caspase-1(1.42±0.30)蛋白表达较对照组NLRP3(0.66±0.15)、ASC(0.41±0.14)和caspase-1(0.70±0.10)显著增加(n t＝5.653，5.602，3.964，均n P<0.01)，而beclin1(0.28±0.09)蛋白表达和LC3II/LC3I(0.69±0.14)比值较对照组beclin1(0.60±0.11)和LC3II/LC3I(1.29±0.23)显著降低(n t＝4.010，3.982，均n P<0.01)。与模型组比较，雷帕霉素组NLRP3(0.80±0.18)和ASC(0.68±0.14)蛋白表达下降(n t＝4.413，3.077，均n P<0.05)，而beclin1(0.65±0.20)蛋白表达和LC3II/LC3I(1.42±0.36)比值增加(n t＝2.965，3.278，均n P<0.05)。n 结论:MPP＋可激活小胶质细胞中的NLRP3炎症小体，并损害细胞的自噬活动；雷帕霉素通过提高自噬活动抑制MPP＋诱导的NLRP3炎症小体激活。“,”Objective:To explore the effect of rapamycin on 1-methyl-4-phenylpyridinium iodide (MPP+ )-induced activation of Nod-like receptor protein 3 (NLRP3) inflammasome in microglia.Methods:The BV2 microglia cells were divided into control group, model group and rapamycin group.The model group and rapamycin group were treated by MPP+ to activate NLRP3 inflammasome, and rapamycin group was pretreated with rapamycin.Quantitative real-time PCR (RT-qPCR) was used to detect the mRNA levels of NLRP3, apoptosis-associated speck-like protein (ASC) and caspase-1.Immunofluorescence was used to detect the protein expression of NLRP3 and interleukin-1β (IL-1β). Western blot was carried out to assess the protein expression of NLRP3, ASC, caspase-1, beclin1 and microtubule-associated protein 1 light chain 3 (LC3).Results:The mRNA levels of NLRP3, ASC and caspase-1 in model group were higher than those in control group (n t=4.825, 3.015, 5.853, all n P<0.05). The mRNA levels of NLRP3 and caspase-1 in rapamycin group were lower than those in model group (n t=2.75, 2.89, both n P<0.05). In model group, the protein expressions of NLRP3 (1.54±0.22), ASC (1.02±0.13) and caspase-1 (1.42±0.30) were higher than NLRP3 (0.66±0.15), ASC (0.41±0.14) and caspase-1 (0.70±0.10) in control group (n t=5.653, 5.602, 3.964, all n P<0.01), while the protein expression of beclin1 (0.28±0.09) and LC3II/LC3I ratio(0.69±0.14) were lower than beclin1 (0.60±0.11) and LC3II/LC3I (1.29±0.23) in control group (n t=4.010, 3.982, both n P<0.01). The protein expressions of NLRP3 (0.80±0.18) and ASC (0.68±0.14) in rapamycin group were lower than those in model group (n t=4.413, 3.077, both n P<0.05), while the protein expression of beclin1 (0.65±0.20) and LC3II/LC3I ratio(1.42±0.36) were higher than those in model group (n t=2.965, 3.278, both n P<0.05).n Conclusion:MPP+ activates NLRP3 inflammasome and impairs autophagic function in microglia.Rapamycin inhibits MPP+ -induced activation of NLRP3 inflammasome by restoring autophagic impairment in microglia.