异氟烷预处理对小鼠脑缺血再灌注后缺血半暗带炎性小体NALP3表达的影响  

Effects of Lsoflurane Preconditioning on the Expressinon of NALP3Inflammasomein Ischemic Penumbra following Cerebral Ischemia-reperfusion in Mice

作　　者：郭辉[1] 陶蕾[1] 孙美艳[1] 于代华[1] 孙绪德[1] 

机构地区：[1]第四军医大学唐都医院麻醉科,陕西西安710038

出　　处：《现代生物医学进展》2015年第23期4467-4470,4480,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81271343;8107099)

摘　　要：目的:研究吸入麻醉药异氟烷预处理对小鼠急性脑梗死缺血半暗带的保护作用,探讨NALP3(NACHT-LRR-PYD-containing Protiein-3 inflammsome)其中发挥的作用。方法:将45只ICR小鼠分为假手术组(Sham)、缺血再灌注组(I/R),异氟烷预处理组(Iso),每组数字15只。其中假手术组仅分离血管,缺血再灌注组采用线栓法制备缺血再灌注小鼠脑缺血半暗带模型,异氟烷预处理组造模前吸入2.0%异氟烷2 h。再灌注24 h后处死小鼠,激光共聚焦检测脑组织内NALP3的表达分布;蛋白免疫印迹(Western blot)法和荧光实时定量PCR(Polymerase Chain Reaction)法检测脑缺血半暗带中NALP3、NF-κB的表达;酶联免疫吸附(ELISA)法检测脑缺血半暗带中IL-1β的表达。结果:脑组织内NALP3的表达主要集中于脑缺血半暗带中,正常组织与梗死区NALP3表达较少;脑缺血再灌注后,I/R组小鼠脑缺血半暗带中NALP3、NF-κB的蛋白表达水平升高,mRNA表达水平显著升高,与sham组相比,差异均有统计学意义(P<0.05);异氟烷与处理后,Iso组小鼠脑缺血半暗带中NALP3、NF-κB的蛋白表达水平降低,mRNA表达水平显著降低,与I/R组相比,差异均有统计学意义(P<0.05);ELISA检测结果显示,I/R组脑缺血半暗带中IL-1β表达水平较Sham组升高了4倍,而异氟烷预处理组的小鼠脑缺血半暗带IL-1β表达水平较I/R组降低36%,差异均具有统计学意义(P<0.05)。结论:异氟烷预处理可抑制NALP3的表达及炎症因子IL-1β的分泌,这种抑制作用可能与异氟烷抑制NF-κB的表达有关。Objective： To explore the expression of NALP3 inflammasome in ischemic penumbra following cerebral ischemia-reperfusion in mice and the effects of Isoflurane preconditioning on it. Methods： ICR mice were randomly divided into three groups（n=15 each）： sham operation group（Sham group）, ischemia-reperfusion group（I/R group）and Isoflurane preconditioning group（Iso group）. The middle cerebral artery of mice in the I/R group were occluded for 2 h, the mice in the Iso group inhaled 2.0% Isoflurane for2 h before their middle cerebral artery were occluded and the shame group just got their vessels dissected bluntly. Confocal laser scanning microscopy was used to detect the expression and distribution of NALP3 in brain. Real-time reverse transcription-polymerase chain reaction（RT-PCR） and Western blot were used to detect the expression of m RNA and protein of NALP3, NF-κB. Enzyme-linked immunosorbent assay（ELISA） was used to examine the expression of interleukin-1β（IL-1β）. Results： NALP3 was found mainly in the ischemic penumbra of I/R group compared with normal cortex and infarcted zone. Compared with sham group, the expressions of NALP3 and NF-κB of I/R group were significantly higher at both protein and m RNA level. The difference was statistically significant（P〈0.05）. Isoflurane preconditioning significantly reduced the expression of NALP3 and NF-κB in Iso group at both proteinandm RNA level than I/R group. The difference was statistically significant（P〈0.05）. The results of ELISA showed that the expression of IL-1β in the brain ischemic penumbra of I/R group was significantly higher than sham groupwhile Isoflurane preconditioning significantly reduced the expression of IL-1β in the brain ischemic penumbra of Iso group than I/R group.The difference was statistically significant（P〈0.05）.Conclusion： Isoflurane preconditioning could suppress the expression of NALP3 and IL-1βin ischemic penumbra probably through inhibiting the expression of NF-κB.

关 键 词：脑缺血再灌注损伤 缺血半暗带 异氟烷预处理 NALP3 NF-κB IL-1Β 

分 类 号：Q953[生物学—动物学] R743.3[医药卫生—神经病学与精神病学]