基于Keap1/Nrf2/NLRP3炎症小体和p62途径探讨右美托咪定改善脑卒中后损伤的内在机制  

Mechanism of dexmedetomidine in improving post-stroke injury based on Keap1/Nrf2/NLRP3 inflammasome and p62 pathways

作　　者：高毅[1] 冷玉芳[2] 张保朝[1] 司小萌 温昌明[1] 孙丽[1] 孙军[1] 张萌[1] 李刚[1] 司海超 张新科[1] 刘展[1] 王飒 GAO Yi;LENG Yufang;ZHANG Baochao;SI Xiaomeng;WEN Changming;SUN Li;SUN Jun;ZHANG Meng;LI Gang;SI Haichao;ZHANG Xinke;LIU Zhan;WANG Sa(Nanyang Central Hospital Affiliated to Zhengzhou University,Nanyang 437000,China)

机构地区：[1]郑州大学附属南阳市中心医院,南阳437000 [2]兰州大学第一附属医院,兰州730099

出　　处：《中国免疫学杂志》2023年第7期1362-1366,共5页Chinese Journal of Immunology

基　　金：河南省重点研发与推广专项计划项目(192102310349);南阳市科技攻关计划项目(KJGG2018082)。

摘　　要：目的:探讨右美托咪定(Dex)对脑卒中后模型脑损伤的改善作用,并基于Keap1/Nrf2/NLRP3炎症小体和p62途径探讨其作用机制。方法:大鼠随机分为假手术组、模型组、依达拉奉组、Dex组,除假手术组外,各组采用线栓法制造缺血性脑卒中大鼠模型,假手术组只穿线不结扎。造模成功后,依达拉奉组大鼠给予依达拉奉3.2 mg/kg,Dex组大鼠灌胃给予Dex 50 mg/kg,模型组、假手术组大鼠给予等体积生理盐水,连续给药14 d。采用改良m-NSS法评价大鼠行为学变化;TTC染色测定脑梗死体积;ELISA检测脑组织中TNF-α、IL-1β、IL-6水平;免疫荧光染色检测脑组织内ROS含量;RT-PCR检测脑组织Keap1、Nrf2、NLRP3、p62 mRNA水平;Western blot检测Keap1、Nrf2、NLRP3、p62蛋白表达。结果:与假手术组相比,模型组大鼠脑神经功能评分明显升高,脑梗死体积明显增大(P<0.01),脑组织中TNF-α、IL-6、IL-1β及ROS水平明显升高(P<0.01),Keap1、Nrf2、NLRP3、p62 mRNA及蛋白表达明显升高(P<0.01);与模型组相比,Dex组大鼠脑神经功能评分明显降低,脑梗死体积明显减小(P<0.01),脑组织中TNF-α、IL-6、IL-1β及ROS水平明显降低(P<0.01),NLRP3、p62 mRNA及蛋白表达明显降低(P<0.01),Keap1、Nrf2 mRNA及蛋白表达明显升高(P<0.01)。结论:Dex可明显改善脑卒中大鼠脑损伤,其可能是通过调控Keap1、Nrf2、NLRP3炎症小体及p62相关基因及蛋白表达实现的。Objective:To investigate the improvement effect of dexmedetomide(Dex)on brain injury in post-stroke injury,and to explore its mechanism based on Keap1/Nrf2/NLRP3 inflammasome and p62 pathways.Methods:Rats were randomly divided into sham group,model group,edaravone group and Dex group,except for sham group,ischemic stroke model was made by thread bolt method in each group.In sham group,only thread was inserted without ligationby.After successful modeling,rats in edaravone group were given edaravone 3.2 mg/kg,rats in Dex group were given 50 mg/kg Dex by gavage,rats in model group and sham group were given equal volume normal saline,continuously for 14 days.Modified m-NSS method was used to evaluate behavioral changes of rats;TTC staining was used to determine volume of cerebral infarction;levels of TNF-α,IL-6,IL-1βin brain tissue were detected by ELISA;ROS content in brain tissue was detected by immunofluorescence staining;mRNA levels of Keap1,Nrf2,NLRP3 and p62 in brain tissue were detected by RT-PCR;protein expressions of Keap1,Nrf2,NLRP3 and p62 were detected by Western blot.Results:Compared with sham group,cerebral nerve function score was significantly increased,cerebral infarction volume was significantly increased(P<0.01),levels of TNF-α,IL-6,IL-1βand ROS in brain tissue were significantly increased(P<0.01),mRNA and protein expressions of Keap1,Nrf2,NLRP3 and p62 were significantly increased in model group(P<0.01).Compared with model group,cerebral nerve function score was significantly decreased,cerebral infarction volume was significantly decreased(P<0.01),levels of TNF-α,IL-6,IL-1βand ROS in brain tissue were significantly decreased,mRNA and protein expressions of NLRP3 and p62 were significantly decreased(P<0.01),mRNA and protein expressions of Keap1 and Nrf2 were significantly increased in Dex group(P<0.01).Conclusion:Dex can significantly improve brain injury of stroke rats,which may be achieved by regulating expressions of Keap1,Nrf2,NLRP3 inflammasome and p62-related genes and proteins.

关 键 词：右美托咪定 脑卒中后损伤 Keap1/Nrf2/NLRP3 P62 

分 类 号：R966[医药卫生—药理学]