摘要
目的脑缺血再灌注损伤机制与炎症反应密切相关,而miRNA-126在血管炎症中起着关键作用。文中旨在探讨miRNA-126通过NLRP3/NF-κB信号轴对脑缺血再灌注损伤小鼠炎症反应的影响及其作用机制。方法将48只雄性小鼠随机分为:假手术组(仅切开颈部皮肤,暴露左颈总动脉后结扎,不插入栓线)、模型组(制备小鼠脑缺血再灌注损伤模型,注射等渗盐水)、阴性对照组(模型建立后侧脑室注射miRNA-126阴性对照miRNA-126 agomir NC)、过表达组(模型建立后侧脑室注射miRNA-126激动剂miRNA-126 agomir),每组12只。进行神经行为学评分,麻醉后处死取小鼠左脑,测定脑组织含水量;采用HE染色,光镜下观察各组小鼠脑皮质组织病理形态学改变;采用ELISA法检测各组小鼠脑组织及血清中IL-1β和IL-6的表达水平;采用Western Blot法检测各组小鼠脑组织中NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、caspase-1、核κB(NF-κB)p65、NF-κB p50、p-NF-κB 65、p-NF-κB 50的蛋白表达水平;采用RT-PCR法检测各组小鼠miRNA-126、NLRP3、ASC、caspase-1、NF-κB p65、NF-κB p50的mRNA表达水平。结果模型组、阴性对照组及过表达组神经行为学评分[(2.33±0.14)、(2.25±0.45)、(1.75±0.22)分]较假手术组(0分)显著升高(P<0.05);过表达组神经行为学评分较阴性对照组、模型组显著降低(P<0.05)。模型组和阴性对照组小鼠脑组织miRNA-126表达水平较假手术组显著降低(P<0.05);过表达组小鼠miR NA-126表达水平较其他3组显著升高(P<0.05)。模型组和阴性对照组小鼠脑组织及血清中IL-1β和IL-6表达水较假手术组显著升高(P<0.05);过表达组IL-1β和IL-6表达水平较模型组和阴性对照组显著降低(P<0.05)。模型组和阴性对照组小鼠脑组织中NLRP3(1.18±0.13、1.25±0.10)、NF-κB p65(1.92±0.11、1.86±0.07)、NF-κB p50 mRNA(0.76±0.06、0.77±0.11)表达水平较假手术组(0.37±0.06、0.53±0.02、0.22±0.01)显著升高
Objective The mechanism of cerebral ischemia-reperfusion injury is highly associated with the inflammatory response.MiRNA-126 plays a key role in vascular inflammation.This study aims to investigate the effect of miRNA-126 on the inflammatory response in mice accompanying cerebral ischemia-reperfusion injury through the NLRP3/NF-κB signal axis,and to explore the mechanisms involved.Methods A total of forty-eight male mice were randomly divided into four groups:the sham-operated group,cerebral ischemia-reperfusion model group,miRNA negative control group(miRNA-126 agomir NC group)and miRNA-126 overexpression group(miRNA-126 agomir group),and each group included twelve mice.The neurobehavioral score was recorded.The left-brain of the mice was sacrificed after anesthesia,and the water content of the brain tissue was measured.HE staining and light microscopy were used to identify the histopathological changes of the cerebral of the mice.The expression levels of inflammatory cytokines IL-1β and IL-6 in brain tissue and serum of mice were detected by ELISA.Western Blot method was used to determine the protein expression levels of NLRP3,ASC,caspase-1,NF-κB p65,NF-κB P50,p-NF-κB 65 and p-NF-κB 50 in brain tissues of mice in each group.RT-PCR was used to test the expression levels of miRNA126,NLRP3,ASC,caspase-1,NF-κB p65 and NF-κB P50 in brain tissue and serum of mice.Results In the sham-operated group,the morphology,and structure of cerebral cortex were normal as healthy mice,being with the dense and orderly arrangement of nerve fibers,with no occurrence of impaired nerve function,and the neurobehavioral score was zero.In both of model group and the miRNA-126 agomir NC group,the ruptured cerebral cortex could be observed visually being with necrotic and disordered cells.The blurred pyknosis and interstitial edema occurred with increased water content of brain tissue.The nerve damage was observed with a significantly increased neurobehavioral score(P<0.05).Compared to the model group,the pathological morphology o
作者
肖莉
熊明敏
王志强
丁鑫
谢馨
XIAO Li;XIONG Ming-min;WANG Zhi-qiang;DING Xin;XIE Xin(Department of Neurology,Western War Zone General Hospital,Chengdu 610083,Sichuan,China;Department of Radiology,Hospital of Chengdu Office of People's Government of Tibetan Autonomous Region,Chengdu 610041,Sichuan,China;Department of Neurology,Affiliated Hospital of Chengdu Medical College,Chengdu 610000,Sichuan,China)
出处
《医学研究生学报》
CAS
北大核心
2020年第4期377-383,共7页
Journal of Medical Postgraduates
