理筋手法干预兔骨骼肌损伤修复中炎症及细胞凋亡的作用机制  

Regulatory mechanism of Lijin manipulation on inflammation and apoptosis in rabbits undergoing skeletal muscle injury repair

作　　者：李开颖 魏晓歌 宋斐 赵振宁 杨楠 李泓钰 赵丽丽 王燕 穆静[1] 马惠昇[1,2,3] Li Kaiying;Wei Xiaoge;Song Fei;Zhao Zhenning;Yang Nan;Li Hongyu;Zhao Lili;Wang Yan;Mu Jing;Ma Huisheng(College of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Ministry of Education Key Laboratory of Modernization of Traditional Medicine for Ethic Minorities in Ningxia,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Ningxia Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of High-Incidence Diseases,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学中医学院,宁夏回族自治区银川市750004 [2]宁夏医科大学宁夏少数民族医药现代化教育部重点实验室,宁夏回族自治区银川市750004 [3]宁夏区域高发病中西医结合防治研究重点实验室,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2025年第20期4215-4222,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(82160944),项目负责人:马惠昇。

摘　　要：背景:骨骼肌细胞的过度凋亡将破坏肌细胞数量的动态平衡,导致骨骼肌病理性损伤。理筋手法治疗骨骼肌损伤疗效确切,但是否能抑制细胞凋亡从而促进损伤骨骼肌的修复尚未可知。目的:探究理筋手法对兔骨骼肌损伤修复过程中减少炎症和细胞凋亡的作用机制。方法:选用45只成年健康日本大耳兔,随机分为空白组、模型组和理筋组。其中空白组不做处理,模型组和理筋组均造成腓肠肌损伤模型;造模后模型组不予治疗,理筋组于第3天开始对损伤腓肠肌进行理筋手法操作,1次/d,15 min/次。各组兔分别于造模后的第7,14,21天取材,苏木精-伊红染色观察腓肠肌一般形态,透射电镜观察腓肠肌超微结构,TUNEL染色观察腓肠肌细胞凋亡情况,ELISA检测腓肠肌白细胞介素1β、白细胞介素6和白细胞介素10的表达,Western blot检测腓肠肌BAX、BCL-2、Caspase3的蛋白表达,RT-PCR检测BAX、BCL-2 mRNA的表达。结果与结论:(1)苏木精-伊红染色结果显示:与模型组比较,各观察点理筋组炎性细胞减少、肌细胞数量增多、肌组织逐渐愈合;(2)透射电镜结果显示:与模型组比较,各观察点理筋组肌纤维排列逐渐整齐、线粒体逐渐完整、Z线排列逐渐趋于规则,游离核糖体聚集;(3)TUNEL染色结果显示:与模型组比较,各观察点理筋组细胞凋亡率逐渐降低(P <0.05);(4)ELISA结果显示,与模型组比较,理筋组白细胞介素1β、白细胞介素6表达持续降低(P <0.05),而白细胞介素10在造模后第7天时升高,随后呈下降趋势(P <0.05);(5)Western blot结果显示:理筋组各观察点BCL-2/BAX蛋白的表达相较于模型组显著升高(P <0.05);Caspase3的蛋白表达均显著降低(P <0.001),且逐渐与空白组相近;(6)RT-PCR结果显示:理筋组各观察点BCL-2/BAX mRNA的表达较模型组均显著升高(P <0.05);(7)结果说明:理筋手法能够抑制炎症、减少细胞凋亡,促进损伤骨骼肌的修复。BACKGROUND:Excessive apoptosis in skeletal muscle cells will destroy the dynamic balance of the number of myocytes,leading to pathological injury of skeletal muscle.Lijin manipulation is effective in treating skeletal muscle injury,but whether it can inhibit apoptosis and promote the repair of skeletal muscle injury is unknown.OBJECTIVE:To explore the mechanism by which Lijin manipulation reduces inflammation and apoptosis during the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group(n=15 per group).No intervention was performed in the blank group.Gastrocnemius muscle percussion molding was performed in both the model group and Lijin group.After modeling,the model group was not treated,while the Lijin manipulation(Stroking,kneading,and rubbing)was performed in the Lijin group on the 3rd day,once a day,15 min/time.Sampling in each group was performed on the 7^(th),14^(th) and 21^(st) days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining.The ultrastructure of gastrocnemius muscle was observed by transmission electron microscopy.Apoptosis of gastrocnemius cells was observed by TUNEL staining.The expressions of interleukin-1β,interleukin-6 and interleukin-10 in gastrocnemius muscle were detected by ELISA.The protein expressions of BAX,BCL-2 and Caspase3 in gastrocnemius muscle were detected by western blot.The mRNA expression of BAX and BCL-2 was detected by RT-PCR.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining results showed that compared with the model group,inflammatory cells decreased in number,myocyte amount increased,and muscle tissue gradually healed in the Lijin group at each observation point.(2)The results of transmission electron microscopy showed that compared with the model group,the arrangement of muscle fibers at each observation point in the Lijin group was gradually orderly,mitochondria were gradually complete,Z-line a

关 键 词：理筋 筋伤 骨骼肌 修复 炎症 细胞凋亡 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R274.3