"三法三穴"推拿对坐骨神经损伤大鼠骨骼肌能量代谢和线粒体动力学的影响OA
Effects of"Three Methods and Three Acupoints"Massage on Energy Metabolism and Mitochondrial Dynamics in Skeletal Muscle of Rats with Sciatic Nerve Injury
目的 从骨骼肌能量代谢和线粒体动力学角度探讨"三法三穴"推拿对坐骨神经损伤(SNI)大鼠运动功能障碍的改善机制.方法 将36只SD大鼠随机分为空白组、假手术组、模型组、推拿组,每组9只.模型组和推拿组采用钳夹法构建SNI模型,假手术组仅暴露坐骨神经不进行夹持,空白组不予干预.分别于造模前及干预10、20次后通过斜板试验评估大鼠运动功能.干预结束后,取大鼠腓肠肌组织,HE染色观察组织形态,比色法测定腓肠肌组织三磷酸腺苷(ATP)含量,免疫荧光染色检测腓肠肌组织线粒体动力学相关因子视神经萎缩症蛋白1(OPA1)、线粒体动力蛋白1(DRP1)、线粒体融合蛋白(MFN)1、线粒体裂变蛋白1(FIS1)表达,实时荧光定量PCR检测腓肠肌组织OPA1、MFN2、DRP1 mRNA表达,Western blot检测腓肠肌组织OPA1、MFN2、DRP1蛋白表达.结果 与空白组和假手术组比较,模型组大鼠斜板试验角度显著减小(P<0.01),表明后肢肌力减弱;腓肠肌纤维排列不规则、间隙增宽,细胞核分布不规律,腓肠肌组织ATP含量显著降低(P<0.01),OPA1、MFN2 mRNA表达显著降低,DRP1 mRNA表达显著升高(P<0.01),OPA1、MFN1、MFN2蛋白表达显著降低,DRP1、FIS1蛋白表达显著升高(P<0.01).与模型组比较,推拿组大鼠斜板试验角度显著增加(P<0.01),表明后肢肌力增强;腓肠肌纤维排列趋于规则、间隙变窄,细胞核分布有序,腓肠肌组织ATP含量显著升高(P<0.01),OPA1、MFN2 mRNA表达显著升高,DRP1 mRNA表达显著降低(P<0.01),OPA1、MFN1、MFN2蛋白表达显著升高,DRP1、FIS1蛋白表达显著降低(P<0.01).结论 "三法三穴"推拿能显著改善SNI大鼠骨骼肌组织结构,恢复其运动功能.其作用机制与调节骨骼肌线粒体动力学失衡状态、提升骨骼肌ATP含量有关.
Objective To analyze the improvement mechanism of motor dysfunction after sciatic nerve injury(SNI)through"three methods and three acupoints"massage from the perspective of energy metabolism and mitochondrial dynamics in skeletal muscle.Methods A total of 36 SD rats were selected and randomly divided into blank group,sham-operation group,model group and massage group,with 9 rats in each group.The SNI model was constructed by clamping method in the model group and massage group;only the sciatic nerve was exposed in the sham-operation group without clamping operation,while the blank group did not receive any treatment.Before modeling,after 10 and 20 times of intervention,the rats were subjected to the inclined plate test to assess their motor functions.After intervention,the gastrocnemius muscle tissue was taken from the rats,and its histomorphology was observed by HE staining,the contents of adenosine triphosphate(ATP)in gastrocnemius muscle tissue was determined by colorimetric assay,immunofluorescence staining was used to detect the expressions of OPA1,DRP1,MFN1 and FIS1 in gastrocnemius muscle tissue,the mRNA expressions of OPA1,MFN2 and DRP1 in gastrocnemius muscle tissue were detected by real-time PCR,and the protein expressions of OPA1,MFN2 and DRP1 in gastrocnemius muscle tissue were detected by Western blot.Results Compared with the blank group and sham-operation group,the angle of the inclined plate test of the rats in the model group was significantly reduced(P<0.01),indicating that the muscle strength of the hind limb was weakened;gastrocnemius muscle fibers arranged irregularly,gaps widened,and nuclei distributed irregularly;the content of ATP in gastrocnemius muscle tissue significantly decreased(P<0.01),the mRNA expressions of OPA1 and MFN2 significantly decreased,the mRNA expression of DRP1 significantly increased,the protein expressions of OPA1,MFN1 and MFN2 significantly decreased(P<0.01),and the protein expressions of DRP1 and FIS1 significantly increased(P<0.01).Compared with the model group,the inclined plate test angle of rats in massage group significantly increased(P<0.01),indicating that the muscle strength of the hind limb was improved;gastrocnemius muscle fibers were more regularly arranged,fiber gaps were narrowed,and the distribution of cell nuclei tended to be uniform,the ATP content in gastrocnemius muscle tissue significantly increased(P<0.01),the mRNA expressions of OPA1 and MFN2 significantly increased,the mRNA expression of DRP1 significantly decreased(P<0.01),the protein expressions of OPA1,MFN1 and MFN2 significantly increased,whereas the protein expressions of DRP1 and FIS1 significantly decreased(P<0.01).Conclusion The"three methods and three acupoints"massage can significantly improve the morphology and structure of the skeletal muscle in SNI rats and recover their motor dysfunction.The mechanism may be closely related to the regulation of mitochondrial dynamics imbalance in skeletal muscle and the enhancement of ATP content in skeletal muscle.
张弘正;许悦;于天源;闫家旺;娜仁图雅;张英琦;刘家玥;张汉钰;孙佳伟
北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488北京中医药大学针灸推拿学院,北京 102488
医药卫生
三法三穴坐骨神经损伤线粒体动力学肌萎缩运动功能障碍大鼠
three methods and three acupointssciatic nerve injurymitochondrial dynamicsmuscular atrophymotor dysfunctionrats
《中国中医药信息杂志》 2026 (1)
70-76,7
国家自然科学基金(82274675)北京市自然科学基金(7232278)
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