低强度激光对运动性肌肉损伤的疗效及其机理研究
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摘要
运动性肌肉损伤是指由不习惯运动,特别是大强度离心运动所诱发的骨骼肌微损伤,人体发生运动性肌肉损伤时,会有延迟出现的肌肉酸痛,因此人体的运动性肌肉损伤也称为延迟性肌肉酸痛。延迟性肌肉酸痛是运动训练过程中常常发生的现象,目前还没有真正有效的治疗方法,低强度激光照射有可能是延迟性肌肉酸痛的一种重要治疗手段,但其疗效尚不确定,作用机制也不清楚。
本研究首次采用动物实验方法调查了不同剂量的低强度激光对运动性肌肉损伤的治疗效果,并探讨了其治疗机理。72只SD大鼠随机分为五个组:(1)安静对照组;(2)运动对照组;(3)三个运动加激光组:低剂量激光组、中剂量激光组、高剂量激光组,运动对照组和运动加激光组又进一步分为运动后24小时亚组和运动后48小时亚组。运动对照组和运动加激光组的大鼠均进行一次力竭性的下坡跑运动,跑台坡度为-16度,跑速为16米/分钟。运动加激光组的大鼠在运动后即刻、18小时、42小时于双侧腓肠肌处接受低强度氦氖激光照射,各剂量组的照射参数分别为12、28和43 J/cm~2 (20、46和71 mW/cm~2, 10 min)。运动后24和48小时进行腓肠肌和血液取材,采用HE染色光镜检查肌组织的病理变化,免疫组化染色光镜检查肌肉结蛋白水平,生物化学方法检测血清肌酸激酶(Creatine Kinase, CK)活性、肌肉超氧化物岐化酶(Superoxide Dismutase, SOD)和一氧化氮合酶(Nitric Oxide Synthase,NOS)活性以及丙二醛(Malondialdehyde, MDA)和一氧化氮(Nitric Oxide, NO)含量。
主要结果如下:
(1)运动对照组的大鼠在离心运动后24和48小时均表现出部分肌纤维坏死,明显的肌肉炎症。与安静对照值相比,其血清CK活性、肌肉MDA水平和NOS活性显著升高,肌肉结蛋白水平显著下降,肌肉SOD/MDA比值和NO/MDA比值明显下降,该两比值在运动后24小时下降更明显,而肌肉炎症和血清CK变化在运动后48小时更明显。
(2)43J/cm~2的激光照射能在离心运动后24和48小时明显减少肌纤维坏死数量,抑制肌肉炎症,显著降低血清CK的升幅,减少肌肉结蛋白的丢失。43 J/cm~2的激光照射在同样时间也显著提高肌肉SOD活性、降低MDA水平以及显著提高NOS活性和NO水平,使肌肉SOD/MDA比值和NO/MDA比值显著上升。该剂量组的运动后各时间血清CK和肌肉MDA值与安静对照水平相比,两者之间无明显差异。
(3)12J/cm~2和28 J/cm~2的激光照射能在离心运动后24和48小时部分抑制肌肉炎症,显著减少肌肉结蛋白的丢失,仅在运动后48小时显著降低血清CK的升幅。对其它指标的作用不明显。
结论:
(1)活性氮氧介质失衡参与了大鼠离心运动性肌肉损伤的发生发展过程。
(2)低强度氦氖激光能够有效地减轻大鼠离心运动性肌肉损伤,促进组织修复,其治疗作用是通过抑制炎症、及时恢复骨骼肌的氧化和抗氧化作用之平衡以及活性氮氧介质之平衡而实现的。
(3)低强度氦氖激光对大鼠离心运动性肌肉损伤的治疗作用是剂量及强度依赖性的。在本文所用的剂量中,43 J/cm~2 (71mW/cm~2, 10min)的剂量照射作用最好。
Exercise-induced muscle damage (EIMD) is referred to the micro-damage of skeletal muscle induced by an unaccustomed, especially high-intensity and eccentric exercise. Human EIMD is also named as delayed onset muscle soreness (DOMS) because the patient with EIMD experienced delayed onset soreness in muscle after exercise. DOMS is a common phenomenon in sports training and so far no really efficient therapeutic approach has been found. Low intensity laser therapy (LLLT) may be an important treatment for DOMS, but the efficacy is uncertain and the mechanism behind the beneficial clinic effects is unclear.
In this study, the effects of low intensity He-Ne laser irradiation at different doses on eccentric exercise-induced muscle injury and its mechanism were investigated with animal experimental method first time. Seventy-two Sprague-Dawley rats were randomly divided into five groups: (1) sedentary control group; (2) exercise control group; (3) three exercise plus laser groups: low-dose laser group, medium-dose laser group, and high-dose laser group. Each of the four exercise groups was further divided into two subgroups: 24 h post-exercise subgroup and 48 h post-exercise subgroup. The exercise was a bout of downhill running (gradient at -16°, speed at 16 m/min) to exhaustion. The laser irradiation was done immediately, 18 h and 42 h after exercise at the middle bellies of bilateral gastrocnemius muscles with He-Ne laser at 12, 28, and 43 J/cm~2(20, 46, and 71 mW/cm~2, 10 min), respectively. At 24 and 48 h after exercise, the gastrocnemius muscles and blood were sampled. Histological change in muscle was examined by HE staining method, and muscle desmin was was examined by immuno-histochemistry staining method, and the analysis for serum creatine kinase (CK), muscle superoxide dismutase (SOD), malondiadehyde (MDA), nitric oxide synthase (NOS) and nitric oxide (NO) were done by biochemistry method.
Main results were as following:
(1) The exercise control group exhibited a few myofibers undergoing necrosis, a marked inflammation in the gastrocnemius muscle 24 and 48 h after exercise. There were significant elevations in serum CK activity, muscle MDA level and NOS activity, and a significant decrease in muscle desmin over sedentary control level in the same post-exercise periods. In addition, there were also obvious reductions in the ratios of SOD/MDA and NO/MDA after exercise, which were more obvious at 24 h after exercise, whereas the muscle inflammation and the elevation of serum CK activity were more obvious at 48 h after exercise.
(2) He-Ne irradiation at 43 J/cm~2 obviously reduced the amount of necrotic myofibers, inhibiting muscle inflammation, and significantly reduced the extent of elevation in serum CK activity and the loss of muscle desmin at both 24 and 48 h after exercise. The irradiation at 43 J/cm~2 also significantly enhanced muscle SOD activity, lowering muscle MDA level, and significantly enhanced muscle NOS activity and NO level so that the ratios of SOD/MDA and NO/MDA obviously increased as compared to the exercise control group in the same post-exercise periods. There were no significant differences in the serum CK activity and muscle MDA level between the laser group receiving 43 J/cm~2 and the sedentary control group at both 24 h and 48 h after exercise.
(3) He-Ne irradiation at 12 or 28 J/cm~2 significantly reduced the loss of muscle desmin and partly inhibited muscle inflammation at both 24 and 48 h after exercise, and significantly inhibited serum CK activity only at 48 h after exercise. The effects of the irradiation at 12 or 28 J/cm~2 on other outcomes were no sigmificant.
Conclusion:
(1) The imbalance between reactive nitric species and reactive oxygen species is involved in the development of eccentric exercise-induced muscle injury in rats.
(2) Low intensity He-Ne laser could efficiently reduce eccentric exercise-induced muscle injury in rats and promote the repair of tissue through inhibiting muscle inflammation and restoring the balance between oxidation and anti-oxidation and the balance between reactive nitric species and reactive oxygen species in muscle in time.
(3) The effects of low intensity He-Ne laser on eccentric exercise-induced muscle injury in rats depend on laser intensity and dose, and the dose 43 J/cm~2 (71 mW/cm~2, 10 min) is the most efficient among the doses used.
本研究首次采用动物实验方法调查了不同剂量的低强度激光对运动性肌肉损伤的治疗效果,并探讨了其治疗机理。72只SD大鼠随机分为五个组:(1)安静对照组;(2)运动对照组;(3)三个运动加激光组:低剂量激光组、中剂量激光组、高剂量激光组,运动对照组和运动加激光组又进一步分为运动后24小时亚组和运动后48小时亚组。运动对照组和运动加激光组的大鼠均进行一次力竭性的下坡跑运动,跑台坡度为-16度,跑速为16米/分钟。运动加激光组的大鼠在运动后即刻、18小时、42小时于双侧腓肠肌处接受低强度氦氖激光照射,各剂量组的照射参数分别为12、28和43 J/cm~2 (20、46和71 mW/cm~2, 10 min)。运动后24和48小时进行腓肠肌和血液取材,采用HE染色光镜检查肌组织的病理变化,免疫组化染色光镜检查肌肉结蛋白水平,生物化学方法检测血清肌酸激酶(Creatine Kinase, CK)活性、肌肉超氧化物岐化酶(Superoxide Dismutase, SOD)和一氧化氮合酶(Nitric Oxide Synthase,NOS)活性以及丙二醛(Malondialdehyde, MDA)和一氧化氮(Nitric Oxide, NO)含量。
主要结果如下:
(1)运动对照组的大鼠在离心运动后24和48小时均表现出部分肌纤维坏死,明显的肌肉炎症。与安静对照值相比,其血清CK活性、肌肉MDA水平和NOS活性显著升高,肌肉结蛋白水平显著下降,肌肉SOD/MDA比值和NO/MDA比值明显下降,该两比值在运动后24小时下降更明显,而肌肉炎症和血清CK变化在运动后48小时更明显。
(2)43J/cm~2的激光照射能在离心运动后24和48小时明显减少肌纤维坏死数量,抑制肌肉炎症,显著降低血清CK的升幅,减少肌肉结蛋白的丢失。43 J/cm~2的激光照射在同样时间也显著提高肌肉SOD活性、降低MDA水平以及显著提高NOS活性和NO水平,使肌肉SOD/MDA比值和NO/MDA比值显著上升。该剂量组的运动后各时间血清CK和肌肉MDA值与安静对照水平相比,两者之间无明显差异。
(3)12J/cm~2和28 J/cm~2的激光照射能在离心运动后24和48小时部分抑制肌肉炎症,显著减少肌肉结蛋白的丢失,仅在运动后48小时显著降低血清CK的升幅。对其它指标的作用不明显。
结论:
(1)活性氮氧介质失衡参与了大鼠离心运动性肌肉损伤的发生发展过程。
(2)低强度氦氖激光能够有效地减轻大鼠离心运动性肌肉损伤,促进组织修复,其治疗作用是通过抑制炎症、及时恢复骨骼肌的氧化和抗氧化作用之平衡以及活性氮氧介质之平衡而实现的。
(3)低强度氦氖激光对大鼠离心运动性肌肉损伤的治疗作用是剂量及强度依赖性的。在本文所用的剂量中,43 J/cm~2 (71mW/cm~2, 10min)的剂量照射作用最好。
Exercise-induced muscle damage (EIMD) is referred to the micro-damage of skeletal muscle induced by an unaccustomed, especially high-intensity and eccentric exercise. Human EIMD is also named as delayed onset muscle soreness (DOMS) because the patient with EIMD experienced delayed onset soreness in muscle after exercise. DOMS is a common phenomenon in sports training and so far no really efficient therapeutic approach has been found. Low intensity laser therapy (LLLT) may be an important treatment for DOMS, but the efficacy is uncertain and the mechanism behind the beneficial clinic effects is unclear.
In this study, the effects of low intensity He-Ne laser irradiation at different doses on eccentric exercise-induced muscle injury and its mechanism were investigated with animal experimental method first time. Seventy-two Sprague-Dawley rats were randomly divided into five groups: (1) sedentary control group; (2) exercise control group; (3) three exercise plus laser groups: low-dose laser group, medium-dose laser group, and high-dose laser group. Each of the four exercise groups was further divided into two subgroups: 24 h post-exercise subgroup and 48 h post-exercise subgroup. The exercise was a bout of downhill running (gradient at -16°, speed at 16 m/min) to exhaustion. The laser irradiation was done immediately, 18 h and 42 h after exercise at the middle bellies of bilateral gastrocnemius muscles with He-Ne laser at 12, 28, and 43 J/cm~2(20, 46, and 71 mW/cm~2, 10 min), respectively. At 24 and 48 h after exercise, the gastrocnemius muscles and blood were sampled. Histological change in muscle was examined by HE staining method, and muscle desmin was was examined by immuno-histochemistry staining method, and the analysis for serum creatine kinase (CK), muscle superoxide dismutase (SOD), malondiadehyde (MDA), nitric oxide synthase (NOS) and nitric oxide (NO) were done by biochemistry method.
Main results were as following:
(1) The exercise control group exhibited a few myofibers undergoing necrosis, a marked inflammation in the gastrocnemius muscle 24 and 48 h after exercise. There were significant elevations in serum CK activity, muscle MDA level and NOS activity, and a significant decrease in muscle desmin over sedentary control level in the same post-exercise periods. In addition, there were also obvious reductions in the ratios of SOD/MDA and NO/MDA after exercise, which were more obvious at 24 h after exercise, whereas the muscle inflammation and the elevation of serum CK activity were more obvious at 48 h after exercise.
(2) He-Ne irradiation at 43 J/cm~2 obviously reduced the amount of necrotic myofibers, inhibiting muscle inflammation, and significantly reduced the extent of elevation in serum CK activity and the loss of muscle desmin at both 24 and 48 h after exercise. The irradiation at 43 J/cm~2 also significantly enhanced muscle SOD activity, lowering muscle MDA level, and significantly enhanced muscle NOS activity and NO level so that the ratios of SOD/MDA and NO/MDA obviously increased as compared to the exercise control group in the same post-exercise periods. There were no significant differences in the serum CK activity and muscle MDA level between the laser group receiving 43 J/cm~2 and the sedentary control group at both 24 h and 48 h after exercise.
(3) He-Ne irradiation at 12 or 28 J/cm~2 significantly reduced the loss of muscle desmin and partly inhibited muscle inflammation at both 24 and 48 h after exercise, and significantly inhibited serum CK activity only at 48 h after exercise. The effects of the irradiation at 12 or 28 J/cm~2 on other outcomes were no sigmificant.
Conclusion:
(1) The imbalance between reactive nitric species and reactive oxygen species is involved in the development of eccentric exercise-induced muscle injury in rats.
(2) Low intensity He-Ne laser could efficiently reduce eccentric exercise-induced muscle injury in rats and promote the repair of tissue through inhibiting muscle inflammation and restoring the balance between oxidation and anti-oxidation and the balance between reactive nitric species and reactive oxygen species in muscle in time.
(3) The effects of low intensity He-Ne laser on eccentric exercise-induced muscle injury in rats depend on laser intensity and dose, and the dose 43 J/cm~2 (71 mW/cm~2, 10 min) is the most efficient among the doses used.
引文
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