摘要
目的:观察智三针电针预刺激对围手术期神经认知障碍(PND)老年大鼠海马组织炎症反应的影响。方法:选择24只老年雄性SD大鼠采用随机数字表法分为对照组、麻醉组、手术组和针刺组,每组6只。针刺组行“智三针”电针预刺激,其他组未行干预。手术组、针刺组采用股骨颈骨折髓内钉内固定术建立老年PND大鼠模型。采用旷场实验、Morris水迷宫实验评价大鼠认知功能;采用ELISA法测定血浆白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α浓度;采用HE染色法观察海马组织细胞形态;采用TUNEL染色法观察海马组织细胞凋亡情况;采用Western blot法和实时荧光定量PCR法分别检测海马组织IL-1β、IL-6、TNF-α蛋白含量及其mRNA表达量。结果:①认知功能:与对照组、麻醉组比较,手术组中央格停留时间、跨格次数、站立次数、游泳总距离、游泳速度和穿越平台次数明显减少,潜伏期、目标象限停留时间百分比明显延长(P<0.05);与手术组比较,针刺组中央格停留时间、跨格次数、站立次数、游泳总距离、游泳速度和穿越平台次数明显增加,潜伏期、目标象限停留时间百分比明显缩短(P<0.05)。②血浆IL-1β、IL-6和TNF-α含量:与对照组比较,麻醉组、手术组、针刺组IL-1β、IL-6和TNF-α含量明显升高(P<0.05);与麻醉组比较,手术组IL-1β、IL-6和TNF-α含量明显升高(P<0.05);与手术组比较,针刺组IL-1β、IL-6和TNF-α含量明显下降(P<0.05)。③海马组织细胞形态和细胞凋亡情况:手术组海马神经元损伤最严重,可见细胞间质水肿,少量胶质细胞增生;针刺组海马神经元损伤轻微,形态较为正常。与对照组、麻醉组比较,手术组、针刺组海马组织凋亡细胞百分比明显升高(P<0.05);与手术组相比较,针刺组海马组织凋亡细胞百分比明显下降(P<0.05)。④海马组织IL-1β、IL-6、TNF-α蛋白含量及其mRNA水平:与对照组、麻醉组比较,手术组IL-1β、IL-6和TNF-α蛋白表达量及其mRNA水平明显升高(P<0.05);与手术组比较,针刺组IL-1β、IL-6和TNF-α蛋白含量及其mRNA水平明显下降(P<0.05)。结论:智三针电针预刺激可减轻PND老年大鼠海马细胞损伤和细胞凋亡,从而改善认知功能,这可能与电针智三针可以抑制老年大鼠海马区IL-1β、IL-6、TNF-α的过度表达,减轻脑组织炎症反应有关。
Objective:To observe the effect of electroacupuncture(EA)prestimulation with mental tri-needles on the inflammatory response of hippocampus in aged rats with perioperative neurocognitive disorders(PND).Methods:A total of 24 aged male SD rats were randomly divided into control group,anesthesia group,operation group and acupuncture group,with 6 cases in each group.The acupuncture group received EA prestimulation with mental tri-needles,and the other groups didn't receive any intervention.The aged PND rat model was established by intramedullary nailing surgery for femoral neck fracture in the operation group and the acupuncture group.Open field experiment and water maze test were used to evaluate the cognitive function;ELISA method was used to detect interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-αin the plasma;HE staining was used to observe the cell morphology of hippocampal tissue;TUNEL staining was used to observe the apoptosis of hippocampal tissue;Western blot and real-time fluorescence quantitative PCR were used to detect the protein content and mRNA expression of IL-1β,IL-6 and TNF-αin hippocampal tissue.Results:(1)Cognitive function:compared with the control group and the anesthesia group,the time of staying in the central quadrant,the number of crossings of the quadrant,the number of standing ups,the total distance of swimming,the swimming speed and the number of platform crossings in the operation group significantly decreased,and the latency and the time of staying in the target quadrant were significantly prolonged(P<0.05);compared with the operation group,the time of staying in the central square,the number of crossings of the quadrant,the number of standing ups,the total distance of swimming,the swimming speed and the number of platform crossings in the acupuncture group significantly increased,and the latency and the time of staying in the target quadrant were significantly shortened(P<0.05).(2)Levels of IL-1β,IL-6 and TNF-αin plasma:compared with the control group,the levels of IL-1β,IL-6 and TNF-αsignificantly increased in the anesthesia group,operation group and acupuncture group(P<0.05);compared with the anesthesia group,the levels of IL-1β,IL-6 and TNF-αin the operation group significantly increased(P<0.05);compared with the operation group,the levels of IL-1β,IL-6 and TNF-αin the acupuncture group significantly decreased(P<0.05).(3)Morphology and apoptosis of hippocampal neurons:the most sever injury of hippocampal neurons occured in the operation group,showing as interstitial edema and a small amount of glial cell proliferation,while the injury of hippocampal neurons was slight and the morphology was relatively normal in the acupuncture group.Compared with the control group and the anesthesia group,the percentages of apoptotic cells in the hippocampus of the operation group and the acupuncture group significantly increased(P<0.05);and compared with the operation group,the percentage of apoptotic cells in the hippocampus of the acupuncture group significantly decreased(P<0.05).(4)Protein contents and mRNA levels of IL-1β,IL-6 and TNF-αin hippocampus:compared with the control group and the anesthesia group,the protein contents and mRNA levels of IL-1β,IL-6 and TNF-αin the operation group significantly increased(P<0.05);compared with the operation group,the protein expression of IL-1β,IL-6 and TNF-αand their mRNA levels in the acupuncture group significantly decreased(P<0.05).Conclusion:EA prestimulation with mental trineedles can reduce the injury and apoptosis of hippocampal cells in aged rats with PND,and improve cognitive function,which may be related to the inhibition of the overexpression of IL-1β,IL-6 and TNF-αin the hippocampus of aged rats and reducing the inflammatory reaction in brain tissue.
作者
王翔锋
黄锦燕
刘威
罗慧琴
王观惠
林芩
陈斌
WANG Xiangfeng;HUANG Jinyan;LIU Wei;LUO Huiqin;WANG Guanhui;LIN Qin;CHEN Bin(The People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350004,China)
出处
《康复学报》
CSCD
2023年第4期325-332,共8页
Rehabilitation Medicine
基金
国家自然科学基金项目(81804175)
福建省卫生健康委员会科研人才培养项目(2018-ZQN-70)
国家中医临床研究基地专项科研课题(JDZX201926)。
关键词
围手术期神经认知障碍
智三针
电针
海马
炎症反应
perioperative neurocognitive disorders
mental tri-needles
electroacupuncture
hippocampus
inflammatory reaction
