摘要
目的观察电针联合康复训练对局灶性脑缺血大鼠NOD样受体蛋白3(NOD-like receptor family pyrin domain containing 3,NLRP3)炎症通路的影响,探讨电针联合康复训练抑制大鼠脑缺血炎症反应的作用机制。方法采用线栓法制备大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型,将45只SD大鼠随机分为假手术组、模型组、电针组、康复训练组、电针联合康复训练组,每组9只。假手术组和模型组不予治疗,电针组电针大鼠“百会”“大椎”穴,康复训练组进行导向性跑台训练,电针联合康复训练组同时予以电针和康复训练干预。比较模型复制完成4 h、7 d后大鼠神经功能评分;苏木精-伊红染色观察缺血侧皮质病理学变化;酶联免疫吸附法检测缺血侧皮质白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)水平;免疫荧光法检测缺血侧皮质中NLRP3、Caspase-1阳性表达水平;析因分析NLRP3、Caspase-1阳性表达水平和IL-1β、IL-18水平的交互性作用。结果模型复制后4 h,各组大鼠的神经功能得分均显著高于假手术组(P<0.05);与模型组比较,电针训练组、康复组、电针联合康复训练组7 d后评分均显著降低(P<0.05)。与假手术组比较,模型组NLRP3、Caspase-1、IL-1β、IL-18表达水平显著升高(P<0.05);与模型组比较,电针组、康复训练组、电针联合康复训练组NLRP3、Caspase-1、IL-1β、IL-18表达水平显著降低(P<0.05);与电针组比较,电针联合康复训练组NLRP3、Caspase-1、IL-1β、IL-18表达水平显著降低(P<0.05);与康复训练组比较,电针联合康复训练组NLRP3、Caspase-1、IL-1β、IL-18表达水平均显著降低(P<0.05)。析因分析提示,电针与康复训练对缺血侧NLRP3通路相关因子的表达具有协同交互作用(P<0.05)。NLRP3、Caspase-1、IL-1β、IL-18均受正向的协同交互作用影响,导致各自水平较之单因素影响而下降更显著。结论电针联合康复训练可通过抑制NLRP3炎症通路,增强抗炎能力,改善神经细胞炎症损伤,达到神经保护作用。
Objective To investigate the effect of electroacupuncture combined with rehabilitation training on the NOD-like receptor family pyrin domain containing 3(NLRP3)inflammatory pathway in rats with focal cerebral ischemia,as well as the mechanism of action of electroacupuncture combined with rehabilitation training in inhibiting inflammatory response in rats with cerebral ischemia.Methods A model of middle cerebral artery occlusion was established.A total of 45 Sprague-Dawley rats were randomly divided into sham-operation group,model group,electroacupuncture group,rehabilitation training group,and electroacupuncture+rehabilitation training group.The rats in the sham-operation group and the model group were not given any treatment;the rats in the electroacupuncture group were given electroacupuncture at Baihui and Dazhui points,those in the rehabilitation training group were given guided treadmill training,and those in the electroacupuncture+rehabilitation training group were given electroacupuncture combined with rehabilitation training.The groups were compared in terms of neurological score at 4 hours and on day 7 after modeling;HE staining was used to observe pathological changes of the cortex at the ischemic side;ELISA was used to measure the levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the cortex at the ischemic side;immunofluorescence assay was used to measure the expression of NLRP3 and caspase-1 in the cortex at the ischemic side;a factorial analysis was performed to investigate the interaction between the expression of NLRP3 and caspase-1 and the levels of IL-1βand IL-18.Results At 4 hours after modeling,each group had a significantly higher neurological score than the sham-operation group(P<0.05),and compared with the model group,the electroacupuncture group,rehabilitation training group,and electroacupuncture+rehabilitation training group had a significant reduction in neurological score on day 7 after modeling(P<0.05).Compared with the sham-operation group,the model group had significant increases in the expression levels of NLRP3,caspase-1,IL-1β,and IL-18(P<0.05);compared with the model group,the electroacupuncture group,rehabilitation training group,and electroacupuncture+rehabilitation training group had significant reductions in the expression levels of NLRP3,caspase-1,IL-1β,and IL-18(P<0.05);compared with the electroacupuncture group,the electroacupuncture+rehabilitation training group had significant reductions in the expression levels of NLRP3,caspase-1,IL-1β,and IL-18(P<0.05);compared with the rehabilitation training group,the electroacupuncture+rehabilitation training group had significant reductions in the expression levels of NLRP3,caspase-1,IL-1β,and IL-18(P<0.05).The factorial analysis showed synergistic interaction between electroacupuncture and rehabilitation training in the expression of NLRP3 pathway-related factors at the ischemic side(P<0.05).NLRP3,caspase-1,IL-1β,and IL-18 were affected by positive synergistic interaction,leading to greater reductions compared with the influence of each factor alone.Conclusion Electroacupuncture combined with rehabilitation training can exert a neuroprotective effect by inhibiting the NLRP3 pathway,enhancing anti-inflammatory ability,and improving inflammatory injury of neural cells.
作者
陶树琴
李梦醒
丁慧
刘箐
毕珂瑶
李斯亮
唐巍
TAO Shuqin;LI Mengxing;DING Hui;LIU Jing;BI Keyao;LI Siliang;TANG Wei(College of Acupuncture and Moxibustion,Anhui University of Chinese Medicine,Anhui Hefei 230012,China)
出处
《安徽中医药大学学报》
CAS
2023年第2期32-37,共6页
Journal of Anhui University of Chinese Medicine
基金
国家自然科学基金项目(81874500)
安徽高校自然科学研究项目(KJ2021A0576,KJ2020A0377)
安徽中医药大学校级科研项目(2021zxts32,2020zryb04)。
关键词
局灶性脑缺血
炎症反应
电针
康复训练
NLRP3通路
Focal cerebral ischemia
Inflammatory response
Electroacupuncture
Rehabilitation training
NLRP3 pathway
