布托啡诺调节PKA/CREB信号通路对缺血性脑卒中大鼠神经元焦亡的影响  

The effect of butorphanol on neuronal pyroptosis in ischemic stroke rats by regulating the PKA/CREB signaling pathway

作　　者：曾恒 仇丽雅 黄青青 ZENG Heng;CHOU Liya;HUANG Qingqing(Department of Anesthesiology,the First People’s Hospital of Guangyuan,Guangyuan,Sichuan 628000,China;Department of Anesthesiology,General Hospital of Western Theater Command,PLA,Chengdu,Sichuan 610000,China)

机构地区：[1]广元市第一人民医院麻醉科,四川省广元市628000 [2]中国人民解放军西部战区总医院麻醉科,四川省成都市610000

出　　处：《中国动脉硬化杂志》2024年第1期17-23,共7页Chinese Journal of Arteriosclerosis

基　　金：广元市指导性科技计划项目(21ZDYF0064)。

摘　　要：[目的]探讨布托啡诺对缺血性脑卒中大鼠神经元焦亡的影响及其在蛋白激酶A(PKA)/环磷酸腺苷(cAMP)反应元件结合蛋白(CREB)通路中的作用。[方法]使用SD大鼠建立缺血性脑卒中大鼠模型,所有大鼠分为对照组、模型组、布托啡诺低剂量组(布托啡诺L组)、布托啡诺高剂量组(布托啡诺H组)、布托啡诺+PKA抑制剂(H-89)组,对大鼠行神经功能评分,TTC染色检测脑梗死体积,HE染色观察脑组织病理特征,铀铅双染色海马区观察神经元焦亡,免疫荧光检测NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体、Caspase-1的表达,酶联免疫吸附法(ELISA)检测白细胞介素1β(IL-1β)、IL-18、cAMP的含量,Western blot检测磷酸化PKA(p-PKA)、PKA、p-CREB、CREB蛋白表达水平。[结果]与对照组比较,模型组大鼠脑组织间隙变大,神经元细胞膜缺损,细胞核核膜凹陷、固缩,神经功能评分升高,脑梗死体积增加,NLRP3、Caspase-1、IL-1β和IL-18水平增加,cAMP、p-PKA/PKA、p-CREB/CREB蛋白表达水平下降(P<0.05);与模型组比较,布托啡诺L、H组脑组织结构较完整,神经元细胞结构异常改善,神经功能评分降低,脑梗死体积减小,NLRP3、Caspase-1、IL-1β和IL-18水平降低,cAMP、p-PKA/PKA、p-CREB/CREB蛋白表达水平升高(P<0.05);与布托啡诺H组比较,布托啡诺+H-89组脑组织细胞空泡变性增加,神经元结构异常,神经功能评分升高,脑梗死体积增加,NLRP3、Caspase-1、IL-1β和IL-18水平增加,cAMP、p-PKA/PKA、p-CREB/CREB蛋白表达水平下降(P<0.05)。[结论]布托啡诺显著抑制缺血性脑卒中大鼠神经元焦亡,可能与激活PKA/CREB信号通路有关。Aim To investigate the effect of butorphanol on neuronal pyroptosis in ischemic stroke rats and its role in protein kinase A(PKA)/cyclic adenosine phosphate(cAMP)response element binding protein(CREB).Methods A rat model of ischemic stroke was established using SD rats.All rats were divided into control group,model group,butorphanol low-dose group(butorphanol L group),butorphanol high-dose group(butorphanol H group)and butorphanol+PKA inhibitor(H-89)group.Neurological function was scored,TTC staining was used to detect cerebral infarction volume,and pathological characteristics of brain tissue were detected by HE staining.Neuronal pyroptosis was observed by uranium lead double staining in hippocampus.The expression of inflammassome NOD-like receptor thermal protein domain associated protein 3(NLRP3)and Caspase-1 were detected by immunofluorescence,and the contents of interleukin-1β(IL-1β),IL-18 and cAMP were detected by enzyme linked immunosorbent assay(ELISA).The expression levels of phosphorylated PKA(p-PKA),PKA,p-CREB and CREB protein were detected by Western blot.Results Compared with the control group,the brain space of rats in the model group was enlarged,the neuron cell membrane was defective,the nucleus and nuclear membrane were sunken and constricted,and the nerve function scores,cerebral infarction volume,NLRP3,Caspase-1,IL-1βand IL-18 levels were increased;the expression levels of cAMP,p-PKA/PKA and p-CREB/CREB protein were decreased(P<0.05).Compared with the model group,the brain structure of butorphanol L and H groups was more complete,the neuronal cell structure was improved,the neural function scores,cerebral infarction volume,NLRP3,Caspase-1,IL-1βand IL-18 levels were decreased,and the protein expression levels of cAMP,p-PKA/PKA and p-CREB/CREB were increased(P<0.05).Compared with butorphanol H group,butorphanol+H-89 group increased vacuolar degeneration of brain tissue,abnormal neuronal structure,neural function scores,cerebral infarction volume,NLRP3,Caspase-1,IL-1βand IL-18 levels;and the

关 键 词：布托啡诺 缺血性脑卒中 蛋白激酶A/环磷酸腺苷反应元件结合蛋白 神经元焦亡 

分 类 号：R7[医药卫生—临床医学] R5