NLRP3炎性小体在缺氧缺血新生大鼠脑组织中的表达及意义  

Expression and significance of NLRP3 inflammatory body in neonatal rats with hypoxic-ischemic brain damage

作　　者：李晓光 王茉琳[2] 罗文哲[2] 逄德志 孙佳斌[2] 张丽华 

机构地区：[1]黑龙江省小儿脑瘫防治疗育中心脑瘫康复科,佳木斯154002 [2]佳木斯大学基础医学院免疫教研室,佳木斯154002

出　　处：《中华行为医学与脑科学杂志》2017年第1期13-16,共4页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：黑龙江省卫生计生委科研课题（2014-264）;佳木斯大学青年项目（Sq2013-019）;佳木斯大学面上项目（JMSUJCM2016-043）;黑龙江省大学生创新创业训练计划项目（201610222051）

摘　　要：目的 探讨新生鼠缺氧缺血性脑损伤（Hypoxic-ischemic brain damage,HIBD）各时间点NLRP3表达情况,寻找空窗期,减轻HIBD组织损伤和功能障碍.方法 96只新生7d龄大鼠随机分为模型组（HIBD）和假手术组（Sham）,每组48只.参照Rice法制备HIBD模型,造模成功后6h、24h、72 h、7d取脑组织,HE染色检测HIBD脑损伤情况,免疫荧光、Western法检测NLRP3、Caspase-1,ELISA法检测IL-1β、IL-18的表达情况.结果 HE染色和免疫荧光结果显示,HIBD组NLRP3蛋白自术后6h显著升高[HIBD组（0.63±0.07）,Sham组（0.43±0.04）],其下游蛋白Caspase-1,IL-1β,IL-18相继活化,分别于术后6 h IL-1β[HIBD组（732.28±108.42） pg/ml,Sham组（584.58±36.35） pg/ml];24 h Caspase-1[HIBD组（0.67±0.09）,Sham组（0.30±0.05）],IL-18[HIBD组（683.84±31.83） pg/ml,Sham组（571.32±50.91） pg/ml]显著增加,差异有统计学意义（P＜0.05）.结论 HIBD时NLRP3信号蛋白及其下游前炎性细胞因子IL-1β、IL-18升高;HIBD组NLRP3蛋白表达变化的时间点早于神经元细胞的缺氧缺血性改变,NLRP3信号可能介导和参与HIBD病程的发生与发展.Objective To investigate the expression of NLRP3 in different time point of HIBD neonatal rats and to search for critical time points and alleviate HIBD dysfunction.Methods 96 newborn rats of 7 days old were randomly divided into HIBD group（n=48） and Sham operation group（n=48）.HIBD model was prepared by referring to Rice method.Brain tissue was taken after 6 h,24 h,72 h,7 d.Brain injury was detected by HE stain.The expression and distribution of NLRP3 and Caspase-1 were detected by immune fluorescence and Western blot,and IL-1β and IL-18 were detected by ELISA.Results HE staining and immunofluorescence showed that NLRP3 protein （HIBD group （0.63±0.07）,Sham group（0.43±0.04）） was increased significantly since 6 h in HIBD group,and its downstream protein Caspase-1,IL-1β and IL-18 were successive activated.The results showed IL-1β （HIBD group（732.28± 108.42）pg/ml,Sham group（584.58± 36.35） pg/ml） was increased significantly since 6 h in HIBD group;Caspase-1 （HIBD group（0.67±0.09）,Sham group（0.30±0.05））,IL-18 （HIBD group（683.84±31.83） pg/ml,Sham group（571.32±50.91） pg/ml） was increased significantly since 24 h in HIBD group（P〈0.05）.Conclusion NLRP3 and its downstream inflammatory cytokines IL-1 β and IL-18 are up-regulated when HIBD occurs.The change of NLRP3protein expression in group HIBD is earlier than changes of neuron.NLRP3 signal may mediate and participate in the occurrence and development of HIBD.

关 键 词：炎症 NLRP3 新生大鼠 脑缺氧缺血 

分 类 号：R742[医药卫生—神经病学与精神病学]