跑台训练脊髓损伤模型大鼠小肠功能恢复与肠道细胞凋亡的关系  

Treadmill training promotes recovery of small intestine function through inhibition of apoptosis in rats with spinal cord injury

作　　者：欧阳率 王先斌 张谦 陈源 邓罗义 王佳 胡寿星 潘晓 吴霜[1,2] Ouyang shuai;Wang Xianbin;Zhang Qian;Chen Yuan;Deng Luoyi;Wang Jia;Hu Shouxing;Pan Xiao;Wu Shuang(Guizhou Medical University,Guiyang 550000,Guizhou Province,China;Department of Rehabilitation Medicine,Affiliated Hospital of Guizhou Medical University,Guiyang 550000,Guizhou Province,China)

机构地区：[1]贵州医科大学,贵州省贵阳市550000 [2]贵州医科大学附属医院康复医学科,贵州省贵阳市550000

出　　处：《中国组织工程研究》2023年第32期5178-5183,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金委员会地区项目(82060419,82260452),项目负责人:吴霜;贵州省科技计划项目(黔科合基础-ZK[2022]重点045),项目负责人:吴霜;贵州省科技计划项目(黔科合基础-ZK[2022]一般438),项目负责人:王先斌;贵州医科大学附属医院博士科研启动基金项目(gyfybsky-2021-5),项目负责人:吴霜;贵州省卫生健康科学技术基金项目(gzwkj2022-030),项目负责人:王先斌。

摘　　要：背景:脊髓损伤后会造成多器官功能障碍,其中肠道功能紊乱严重影响患者疾病治疗与生活质量,有研究表明有氧运动可以影响肠道结构与功能,但机制尚不明确。目的:探讨跑台训练对脊髓损伤大鼠小肠功能的影响及其与肠道细胞凋亡的关系。方法:实验采用改良脊髓打击法制备大鼠T10不完全脊髓损伤模型。54只SD雌性大鼠按随机数字法分为假手术组、脊髓损伤组和脊髓损伤运动组各18只。脊髓损伤运动组在造模后7 d开始跑台运动训练,总时长3周;在末次干预后,采用BBB评分和斜板实验评估运动能力,取脊髓损伤处上下0.5 cm的脊髓组织检测神经元存活情况和星形胶质细胞活化水平以评估运动对神经功能的影响;采用灌胃染色剂法检测各组大鼠的首粒黑便排出时间、肠道染色推进效率以评估肠道转运功能;分别于脊髓损伤后14,21,28 d,采用苏木精-伊红染色观察小肠组织的病理结构变化,免疫荧光检测小肠紧密连接蛋白Occludin的表达,Western blot检测凋亡相关蛋白Bcl-2、Caspase-9、Bax、Caspase-3和Cyt-C的表达。结果与结论:①与脊髓损伤组比较,脊髓损伤运动组BBB评分、斜板角度均较高(P<0.05),神经元存活更多,神经炎症较轻;②在脊髓损后14 d,与假手术组相比,脊髓损伤组和脊髓损伤运动组小肠组织出现黏膜层破坏、肠绒毛萎缩、杯状细胞数量减少、核固缩等病理改变,脊髓损伤组小肠病理损伤在21 d达到高峰并持续到实验结束,脊髓损伤运动组大鼠小肠组织黏膜连续性、肠绒毛萎缩等病理改变在术后28 d有明显改善;③脊髓损伤运动组首粒黑便排出时间低于脊髓损伤组(P<0.05)、肠道推进效率高于脊髓损伤组(P<0.05),提示肠道运输功能好转;④脊髓损伤28 d,与脊髓损伤组相比,脊髓损伤运动组的小肠紧密连接蛋白Occludin表达升高(P<0.01),凋亡相关蛋白Caspase-3、Caspase-9、Bax、Cyt-C表达降�BACKGROUND:Spinal cord injury can cause multi-organ dysfunction,among which intestinal dysfunction seriously affects patients’disease treatment and quality of life.Some studies have shown that aerobic exercise can improve intestinal structure and function,but the mechanism needs to be further explored.OBJECTIVE:To investigate the effect of treadmill training on the function of small intestine in rats with spinal cord injury and its relationship with intestinal cell apoptosis.METHODS:The modified spinal cord percussion method was used to prepare a rat model of T10 incomplete spinal cord injury.A total of 54 female Sprague-Dawley rats were randomly divided into a sham-operated group,a model group,and a training group,with 18 rats in each group.The treadmill training was started 7 days after modeling for 3 weeks.After the final intervention,the Basso-Beattie-Bresnahan score and the inclined plate test were used for locomotor testing.Spinal cord tissue samples were taken 0.5 cm above and below the injured site to detect neuronal survival and astrocyte activation levels for testing the effect of exercise on neurological function.The intestinal transport function was evaluated by gavage staining of the first black stool and intestinal staining propulsive efficiency in each group.At 14,21 and 28 days after spinal cord injury,the pathological structural changes of the small intestine were observed by hematoxylin-eosin staining and the expression of the small intestinal tight junction protein Occludin was detected by immunofluorescence;the expression levels of apoptosis-related proteins Bcl-2,Caspase-9,Bax,Caspase-3,and Cyt-C were detected by western blot.RESULTS AND CONCLUSION:Compared with the model group,the training group had recovered neurological function with higher Basso-Beattie-Bresnahan scores and inclined plate angles(P<0.05),more survived neurons,and less neuroinflammation.At 14 days after spinal cord injury,compared with the sham-operated group,the small intestinal tissue in the model and training groups sh

关 键 词：脊髓损伤 跑台 肠道功能 紧密连接蛋白 细胞凋亡 

分 类 号：R496[医药卫生—康复医学] R319[医药卫生—临床医学] R455