基于腺苷酸激活蛋白激酶-自噬信号通路探讨减重平板训练对脊髓损伤大鼠运动功能的影响  

Effect of Body Weight Supported Treadmill Training on Motor Function of Rats with Spinal Cord Injury Based on AMPK-Autophagy Signaling Pathway

作　　者：蔡吉利 翟晨元 王尊 伍琦 方露 王钰 王彤[4] CAI Jili;ZHAI Chenyuan;WANG Zun;WU Qi;FANG Lu;WANG Yu;WANG Tong(School of Rehabilitation Medicine,Nanjing Medical University,Nanjing,Jiangsu 210029,China;School of Acupuncture-Moxibustion and Tuina,School of Health Preservation and Rehabilitation,Nanjing University of Chinese Medi-cine,Nanjing,Jiangsu 210023,China;The First Affiliated Hospital of University of South China,Hengyang,Hunan 421001,China;The First Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210029,China)

机构地区：[1]南京医科大学康复医学院,江苏南京210029 [2]南京中医药大学针灸推拿学院·养生康复学院,江苏南京210023 [3]南华大学衡阳医学院附属第一医院,湖南衡阳421001 [4]南京医科大学第一附属医院,江苏南京210029

出　　处：《康复学报》2024年第4期370-376,共7页Rehabilitation Medicine

基　　金：国家自然科学基金面上项目(82172541)。

摘　　要：目的 基于腺苷酸激活蛋白激酶(AMPK)-自噬信号通路探讨减重平板训练对脊髓损伤(SCI)大鼠运动功能的影响及其作用机制。方法 选择30只10周龄SPF级雌性SD大鼠,按照随机数字表法分为假手术组、模型组和运动组,每组10只。模型组和运动组均采用Allen's法制作T10不完全性脊髓损伤模型,假手术组仅进行假手术。3组均于SCI造模前3 d进行预适应运动训练,以适应跑台环境。运动组于造模后第8天进行减重平板训练,减重为大鼠体质量的20%~40%,跑台速度6 m/min,20 min/次,2次/d,持续训练3周;假手术组和模型组造模后均自由活动,不进行减重平板训练。分别在造模后第1、7、14、21、28天采用脊髓损伤行为学(BBB)评分法评估各组大鼠后肢运动功能;于干预结束后(SCI后第28天)采用步态分析处理系统评估大鼠的步态参数(足印面积、步幅长度、触地强度、摆动速度和运动速度);采用尼氏染色法观察脊髓神经元形态及数目;采用Western blot法检测脊髓组织p-AMPK、t-AMPK、LC3、p62蛋白表达水平。结果(1) BBB评分:与假手术组比较,模型组、运动组造模后第1、7、14、21、28天BBB评分明显更低,差异均具有统计学意义(P<0.05);与模型组同一时间点比较,运动组造模后14、21、28天BBB评分明显更高,差异均具有统计学意义(P<0.05);模型组、运动组内不同时间点BBB评分比较,差异均具有统计学意义(P<0.05)。(2)步态参数:与假手术组比较,模型组足印面积、触地强度、摆动速度、步幅长度和运动速度均明显更低,运动组足印面积明显更低,差异均具有统计学意义(P<0.05);与模型组比较,运动组第28天足印面积、触地强度、摆动速度、步幅长度和运动速度均明显更高,差异均具有统计学意义(P<0.05)。(3)脊髓前角神经元形态及数目:与假手术组比较,模型组脊髓前角神经元数目明显减少(P<0.05),可见细胞核固缩。与模型组比较,Objective To explore the effect and mechanism of body weight supported treadmill training on motor function of rats with spinal cord injured(SCI)based on the adenosine monophosphate-activated protein kinase(AMPK)-autophagy signaling pathway.Methods A total of 3010-week-old female SD rats were randomly divided into sham operation group,model group,and exercise group,with 10 cases in each group.Allen's method was used to establish T10 incomplete spinal cord injury models in the model group and the exercise group,while the sham operation group only underwent sham operation.All the three groups received preconditioning exercise training for 3 days before SCI modeling to adapt to the treadmill environment.From the 8th day after modeling,the exercise group received treadmill training with weight reduction of 20%-40%of the body mass,and the treadmill speed was set at 6 m a minute,20 minutes a time,twice a day for 3 weeks.The sham operation group and the model group were allowed to move freely without treadmill training after modeling.Basso-Beattie-Bresnahan(BBB)method was used to assess motor function of hind limbs of rats at the 1st,7th,14th,21st and 28th days after modeling.Gait analysis system was used to assess gait parameters(footprint area,stride length,touchdown intensity,swing speed and movement speed)after intervention(at the 28th day after SCI).Nissl staining was used to observe the morphology and count of spinal cord neurons.Western blot method was used to detect the protein expression levels of p-AMPK,t-AMPK,LC3 and p62 in spinal cord tissue.Results(1)BBB score:compared with the sham operation group,the BBB score of the model group and the exercise group were significantly lower at the 1st,7th,14th,21st and 28th days after modeling(P<0.05).Compared with the model group at the same time,the BBB score of the exercise group was significantly higher at the 14th,21st and 28th days after modeling,and the differences were statistically significant(P<0.05).The BBB score in the model group and the exercise group were

关 键 词：脊髓损伤 减重平板训练 腺苷酸激活蛋白激酶 自噬 运动功能 神经功能 

分 类 号：R651.2[医药卫生—外科学] R493[医药卫生—临床医学]