中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (32): 5178-5183.doi: 10.12307/2023.846

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

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

欧阳率1，王先斌1，2，张  谦2，陈  源2，邓罗义2，王  佳1，胡寿星1，潘  晓1，吴  霜1，2   

  1. 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
  • 收稿日期:2022-10-21 接受日期:2022-11-30 出版日期:2023-11-18 发布日期:2023-03-23
  • 通讯作者: 吴霜，教授，博士，主任医师，贵州医科大学，贵州省贵阳市 550000；贵州医科大学附属医院康复医学科，贵州省贵阳市 550000
  • 作者简介:欧阳率，男，1995年生，贵州省凯里市人，侗族，贵州医科大学在读硕士，主要从事肌骨康复研究。
  • 基金资助:
    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

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

Ouyang shuai1, Wang Xianbin1, 2, Zhang Qian2, Chen Yuan2, Deng Luoyi2, Wang Jia1, Hu Shouxing1, Pan Xiao1, Wu Shuang1, 2   

  1. 1Guizhou Medical University, Guiyang 550000, Guizhou Province, China; 2Department of Rehabilitation Medicine, Affiliated Hospital of Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • Received:2022-10-21 Accepted:2022-11-30 Online:2023-11-18 Published:2023-03-23
  • Contact: Wu Shuang, MD, Professor, Chief physician, Department of Rehabilitation Medicine, Affiliated Hospital of Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • About author:Ouyang Shuai, Master candidate, Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China (Regional Program), Nos. 82060419 and 82260452 (both to WS); Guizhou Provincial Science and Technology Project, No. ZK[2022] key 045 (to WS); Guizhou Provincial Science and Technology Project, No. ZK[2022] general 438 (to WXB); The Doctoral Research Foundation of the Affiliated Hospital of Guizhou Medical University, No. gyfybsky-2021-5 (to WS); Guizhou Provincial Health Commission Foundation, No. gzwkj2022-030 (to WXB)

PDF

106

摘要：


文题释义：
脊髓损伤后肠道功能紊乱：脊髓损伤后会造成多器官功能障碍，其中小肠功能紊乱严重影响机体营养吸收、代谢和免疫，肠道系统与神经系统之间存在非常紧密且复杂的双向交流通路，其作用机制需要深入探索。
紧密连接蛋白：是肠道维持肠道屏障功能的结构蛋白，研究表明运动可以改变肠道结构和功能，运动、肠道和细胞凋亡三者之间可能存在关联。

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表达降低(P < 0.05)、Bcl-2表达升高(P < 0.05)；⑤结果表明，跑台运动训练可保护脊髓损伤大鼠神经和小肠系统，其中小肠功能的恢复可能与小肠组织细胞凋亡水平有关。 
https://orcid.org/0000-0002-4442-330X(欧阳率)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 脊髓损伤, 跑台, 肠道功能, 紧密连接蛋白, 细胞凋亡

Abstract: 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 showed pathological changes, such as mucosal layer disruption, intestinal villus atrophy, reduced number of cup cells, and nuclear consolidation. The pathological damage of the small intestine in the model group peaked at 21 days after spinal cord injury and lasted until the end of the experiment. The pathological changes such as mucosal continuity and intestinal villus atrophy in the training group showed significant improvement at 28 days after surgery. The first black stool time in the training group was lower than that in the model group (P < 0.05), and the intestinal propulsive efficiency was higher than that in the model group (P < 0.05), suggesting that the intestinal transport function improved. At 28 days after spinal cord injury, the expression of the small intestinal tight junction protein Occludin was higher in the training group than the model group (P < 0.01), as well as the Bcl-2 expression (P < 0.05) but the expression of apoptosis-related proteins Caspase-3, Caspase-9, Bax Caspase-9, Bax, and Cyt-C was decreased (P < 0.05). To conclude, the treadmill training can protect the nervous system and small intestinal system in rats with spinal cord injury and the recovery of small intestine function may be related to the level of apoptosis in small intestine tissue.

Key words: spinal cord injury, treadmill, intestinal function, tight junction protein, apoptosis

中图分类号: