推拿对延迟性肌肉酸痛的恢复及对大鼠氧化/抗氧化平衡的影响
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摘要
本论文包括文献研究和动物研究两大部分。
文献研究部分:延迟性肌肉酸痛(Delayed Onset Muscle Soreness,DOMS)是指在激烈运动后8-24h所发生的肌肉酸痛现象,通常在运动后24~72h达到最到最高点,并持续数天才会完全恢复。DOMS好发于不习惯、大强度的离心训练之后,在训练结束数小时发作,并持续数日。它是运动训练中的常见症状,以离心训练结束后延迟发作的疼痛和肌力减退为主要表现。由于推拿方法在各运动队的广泛运用及其独特的优势,所以在DOMS的防治上越来越受重视。此部分研究回顾了西医学的相关研究进展,对DOMS的发生机制、评定及防治措施进行了总结,系统整理了中医学对DOMS的认识,从中医学和西医学的角度系统阐述推拿疗法防治运动损伤的机制,为推拿防治DOMS提供理论依据,为动物实验的设计、确定有效的观察指标、制定完善的实验方法奠定了基础。
动物实验研究部分:目的:通过建立大鼠DOMS模型,对大鼠血清乳酸(LD)、乳酸脱氢酶(LDH)、肌酸激酶(CK)、丙二醛(MDA)、超氧化物歧化酶(SOD)、羟自由基(·OH)等生化指标的观察和检测,研究推拿治疗对DOMS的恢复及对大鼠氧化/抗氧化平衡的影响。方法:采用鼠龄6周的健康雄性SD大鼠130只,按照随机原则将动物分为安静组(A)、运动对照组(B)及推拿组;推拿组分为运动前推拿组(C)、运动后推拿组(D),推拿组、运动对照组又分别随机分为1h组、12h组、24h组、48h组,各组10只。每日上午7点至10点时进行实验。运动对照组让实验大鼠在100 cm×60 cm×70 cm的玻璃缸中负重游泳,静水深55cm,水温(33±2)℃,负重为大鼠体重的3%,连续下沉三次即将动物取出游泳缸用电吹风吹干入笼。运动前推拿组与运动后推拿组分别在运动前或运动后将大鼠固定于大鼠固定架上,由专人运用捏法和捻法施于大鼠的两侧下肢,两侧下肢各5min,每只大鼠推拿10min。运动对照组及推拿组在大鼠游泳运动结束后1h、12h、24h和48h分别进行取样;安静组在48h进行取样。用大鼠专用断头铡刀将大鼠断头处死,然后迅速将血液收集于离心管中,取血清送检。比较各组不同时段各生化指标的变化。结果:(1)与安静组比较,运动对照组、运动前推拿组及运动后推拿组大鼠血清LD均有不同程度升高,且有显著性差异;在运动对照组,运动后12h大鼠血清LD达到峰值,随后24~48h逐渐降低;运动前推拿1h组大鼠血清LD显著下降,12~24h组大鼠血清LD逐渐升高至峰值;运动后推拿1h组大鼠血清LD明显降低,12h升高达到峰值,随后下降,运动后推拿1h及24h组大鼠血清LD明显低于运动前推拿1h及24h组水平。(2)运动后大鼠血清LDH立即升高,在运动后12h达到峰值,随后逐渐下降,至运动后48h低于安静组水平;运动对照组及推拿组LDH与运动对照组及推拿组各相同时相的LD的变化基本一致。伴随血清LD的降低,血清LDH的活性也同步降低。(3)运动后血清CK立即升高,与安静组相比,运动对照组、运动前推拿组及运动后推拿组大鼠血清CK均有不同程度升高。与运动对照1h与12h组相比较,运动前推拿1h与12h组大鼠血清CK无显著性差异,运动后推拿1h与12h组大鼠血清CK则明显升高,两者相比有显著性差异;与运动前推拿1h、12h及48h组相比较,运动后推拿各相同时相大鼠血清CK明显升高,两者相比有显著性差异。(4)运动对照组大鼠血清·OH的含量明显升高,在运动后1h立即升高达到峰值;运动前或运动后推拿1h与12h组大鼠血清·OH的含量明显下降,与运动对照1h与12h组相比有显著性差异;运动前推拿1h、12h及24h组大鼠血清·OH的含量与运动后推拿相同时相比较无显著性差异。(5)运动能使大鼠血清MDA明显升高,运动对照1h、12h、24h、48h组MDA水平,与安静组比较有显著性差异;运动前推拿1h、12h、24h组MDA与安静组比较稍有增加,两者相比,无显著性差异;运动后推拿除1h组MDA有所升高,12h、24h、48h组MDA水平均达到安静组水平。(6)运动前或运动后推拿1h、12h、24h、48h组大鼠血清SOD明显高于运动对照组各相同时相水平,与安静组相比较也有显著性差异。(7)与安静组相比较,运动对照组SOD/MDA的比值显著降低;运动前推拿1h、12h、24h组SOD/MDA的比值与安静组相比无显著性差异;与运动对照1h、12h、24h及48h组相比,运动前推拿与运动后推拿组各相同时段均有显著升高;在运动前推拿组,SOD/MDA的比值在运动后1h、12h、24h均升高,至48h下降低于安静组水平;运动后推拿组SOD/MDA的比值在推拿后呈逐渐升高的趋势,至推拿后48h达到峰值。结论:(1)剧烈运动后模型大鼠血清LD的变化与运动疲劳有关,与DOMS无关。(2)运动前推拿能有效防治离心运动对肌肉的损伤。(3)运动前推拿或运动后推拿均能降低模型大鼠血清·OH和MDA的生成,提高SOD活性和SOD/MDA的比值,运动前推拿能更有效地抑制模型大鼠血清·OH和MDA的生成,表明推拿治疗具有良好的抗脂质过氧化作用。(4)本研究重点观察了运动前推拿与运动后推拿对DOMS模型大鼠在恢复期的不同时相血清中LD、LDH、CK、·OH、MDA和SOD的变化规律;研究结果表明,运动前推拿能降低血清内CK和LDH活性,运动前推拿或运动后推拿能抑制血清中·OH与MDA的生成,提高血清中SOD的活性和SOD/MDA的比值,间接说明推拿能延缓大鼠DOMS的产生,加速肌肉酸痛的消除,有效控制脂质过氧化损伤,提高机体抗氧化能力,对肌细胞具有良好的保护作用。
This dissertation includes two parts, i.e. literature research and animal experiment. Literature research
Delayed-onset muscle soreness (DOMS) describes muscle pain and tenderness that typically develop several hours postexercise and consist of predominantly eccentric muscle actions, especially if the exercise is unfamiliar. As we know, traditional Chinese Tuina is widely used in sports teams with its unique advantages in relieving exercise-induced fatigue and strengthening sporting abilities, and it has gained more and more attention in prevention and treatment of DOMS. This part reviews the relative literatures in western and traditional medicine, and sums up the pathogenesis and evaluation of DOMS, and approaches to prevention and treatment for DOMS. This part also covers the mechanism of Tuina on preventing and treating sports injuries, which provides theoretical basis for prevention and treatment on DOMS by Tuina. The design of animal experiment, observing data and experimental method were established on the basis of these literatures.
Animal experiment research
Objective: to evaluate the effects of Tuina on DOMS and the relationship of oxidative-antioxidative balance disturbance in rats by testing serum LD, LDH, CK, MDA,SOD、·OH.
Method
130 healthy and clean male SD rats of six-week old were randomly divided into 4 groups including normal control group(A), exercise control group(B), Tuina pre-exercise group(C) and Tuina post-exercise(D). Group B, C and D were randomly divided into 1h, 12h, 24h and 48h groups. Each group consists of 10 rats. Group B were put into a glass pool of 100 cm×60 cm×70 cm to swim carrying a subject of 3% its body weight till exhausted. Group C and Group D were treated 10 minutes by Tuina pre-exercise or post-exercise. The rats in Group B, C and D were killed to take blood sample 1h, 12h, 24h and 48h post-exercise or after Tuina. While the normal control group were killed after 48h to take blood sample.
Results
(1) In comparison with the normal control group, the level of serum LD in exercise control group, Tuina pre-exercise group and Tuina post-exercise rose significantly. In comparison with the exercise control group, the level of serum LD in Tuina pre-exercise group and Tuina post-exercise lowered significantly. (2) The level of serum LDH rose immediately to peak after 12h postexercise. The level of serum LDH lowered in level with LD in Group B, C and D. (3) In comparison with the normal control group, the level of serum CK in group B, group C and group D rose immediately. In comparison with group B, the level of serum CK in 1h, 12h in group C had no significant difference while it rose significantly in 1h, 12h in group D. In comparison with group C, the level of serum CK after 1h,12h and 48h in group D rose signifieantly. (4) The level of serum·OH rose immediately to peak after 1h in group B. In comparison with group B after 1h,12h, the level of serum·OH lowered signifieantly in group C and group D. The level of serum OH in 1h, 12h, and 24h had no significant difference between group C and group D. (5) The level of serum MDA rose signifieantly in group B. In comparison with group A, the level of serum MDA in 1h,12h and 24h in group C had no significant difference. While the level of serum MDA after 12h, 24h and 48h in group D had no significant difference. (6) In comparison with group A and group B, the level of serum SOD rose signifieantly in 1h, 12h, 24h and 48h in both group C and group D. (7) In comparison with group A, the ratio of SOD/MDA lowered significantly in group B. In comparison with group B, the ratio of SOD/MDA rose signifieantly in 1h, 12h, 24h and 48h in both group C and group D.
Conclusion
(1) The change of serum LD is related with exercise-induced fatigue, but it is not related with DOMS. (2) Tuina pre-exercise is effective in alleviating muscle damage caused by eccentric exercise in DOMS. (3) Tuina pre-exercise or postexercise are effective in lowering the level of serum OH and MDA, while raising the activity of serum SOD and the ratio of SOD/MDA. Tuina pre-exercise is more effective. (4) This experiment focused on Tuina pre-exercise or postexercise on rats of DOMS in testing the changes of LD、LDH、CK、·OH、MDA and SOD. The experimental result shows that Tuina pre-exercise is effective in lowering the level of serum LDH and CK. Tuina pre-exercise or postexercise are effective in lowering the level of serum·OH and MDA, while raising the activity of serum SOD and the ratio of SOD/MDA. Tuina is effective in delaying DOMS, reducing muscle soreness, controlling the damage of lipid peroxidation, increasing the ability in anti-oxidation and has good protection to muscle cell.
文献研究部分:延迟性肌肉酸痛(Delayed Onset Muscle Soreness,DOMS)是指在激烈运动后8-24h所发生的肌肉酸痛现象,通常在运动后24~72h达到最到最高点,并持续数天才会完全恢复。DOMS好发于不习惯、大强度的离心训练之后,在训练结束数小时发作,并持续数日。它是运动训练中的常见症状,以离心训练结束后延迟发作的疼痛和肌力减退为主要表现。由于推拿方法在各运动队的广泛运用及其独特的优势,所以在DOMS的防治上越来越受重视。此部分研究回顾了西医学的相关研究进展,对DOMS的发生机制、评定及防治措施进行了总结,系统整理了中医学对DOMS的认识,从中医学和西医学的角度系统阐述推拿疗法防治运动损伤的机制,为推拿防治DOMS提供理论依据,为动物实验的设计、确定有效的观察指标、制定完善的实验方法奠定了基础。
动物实验研究部分:目的:通过建立大鼠DOMS模型,对大鼠血清乳酸(LD)、乳酸脱氢酶(LDH)、肌酸激酶(CK)、丙二醛(MDA)、超氧化物歧化酶(SOD)、羟自由基(·OH)等生化指标的观察和检测,研究推拿治疗对DOMS的恢复及对大鼠氧化/抗氧化平衡的影响。方法:采用鼠龄6周的健康雄性SD大鼠130只,按照随机原则将动物分为安静组(A)、运动对照组(B)及推拿组;推拿组分为运动前推拿组(C)、运动后推拿组(D),推拿组、运动对照组又分别随机分为1h组、12h组、24h组、48h组,各组10只。每日上午7点至10点时进行实验。运动对照组让实验大鼠在100 cm×60 cm×70 cm的玻璃缸中负重游泳,静水深55cm,水温(33±2)℃,负重为大鼠体重的3%,连续下沉三次即将动物取出游泳缸用电吹风吹干入笼。运动前推拿组与运动后推拿组分别在运动前或运动后将大鼠固定于大鼠固定架上,由专人运用捏法和捻法施于大鼠的两侧下肢,两侧下肢各5min,每只大鼠推拿10min。运动对照组及推拿组在大鼠游泳运动结束后1h、12h、24h和48h分别进行取样;安静组在48h进行取样。用大鼠专用断头铡刀将大鼠断头处死,然后迅速将血液收集于离心管中,取血清送检。比较各组不同时段各生化指标的变化。结果:(1)与安静组比较,运动对照组、运动前推拿组及运动后推拿组大鼠血清LD均有不同程度升高,且有显著性差异;在运动对照组,运动后12h大鼠血清LD达到峰值,随后24~48h逐渐降低;运动前推拿1h组大鼠血清LD显著下降,12~24h组大鼠血清LD逐渐升高至峰值;运动后推拿1h组大鼠血清LD明显降低,12h升高达到峰值,随后下降,运动后推拿1h及24h组大鼠血清LD明显低于运动前推拿1h及24h组水平。(2)运动后大鼠血清LDH立即升高,在运动后12h达到峰值,随后逐渐下降,至运动后48h低于安静组水平;运动对照组及推拿组LDH与运动对照组及推拿组各相同时相的LD的变化基本一致。伴随血清LD的降低,血清LDH的活性也同步降低。(3)运动后血清CK立即升高,与安静组相比,运动对照组、运动前推拿组及运动后推拿组大鼠血清CK均有不同程度升高。与运动对照1h与12h组相比较,运动前推拿1h与12h组大鼠血清CK无显著性差异,运动后推拿1h与12h组大鼠血清CK则明显升高,两者相比有显著性差异;与运动前推拿1h、12h及48h组相比较,运动后推拿各相同时相大鼠血清CK明显升高,两者相比有显著性差异。(4)运动对照组大鼠血清·OH的含量明显升高,在运动后1h立即升高达到峰值;运动前或运动后推拿1h与12h组大鼠血清·OH的含量明显下降,与运动对照1h与12h组相比有显著性差异;运动前推拿1h、12h及24h组大鼠血清·OH的含量与运动后推拿相同时相比较无显著性差异。(5)运动能使大鼠血清MDA明显升高,运动对照1h、12h、24h、48h组MDA水平,与安静组比较有显著性差异;运动前推拿1h、12h、24h组MDA与安静组比较稍有增加,两者相比,无显著性差异;运动后推拿除1h组MDA有所升高,12h、24h、48h组MDA水平均达到安静组水平。(6)运动前或运动后推拿1h、12h、24h、48h组大鼠血清SOD明显高于运动对照组各相同时相水平,与安静组相比较也有显著性差异。(7)与安静组相比较,运动对照组SOD/MDA的比值显著降低;运动前推拿1h、12h、24h组SOD/MDA的比值与安静组相比无显著性差异;与运动对照1h、12h、24h及48h组相比,运动前推拿与运动后推拿组各相同时段均有显著升高;在运动前推拿组,SOD/MDA的比值在运动后1h、12h、24h均升高,至48h下降低于安静组水平;运动后推拿组SOD/MDA的比值在推拿后呈逐渐升高的趋势,至推拿后48h达到峰值。结论:(1)剧烈运动后模型大鼠血清LD的变化与运动疲劳有关,与DOMS无关。(2)运动前推拿能有效防治离心运动对肌肉的损伤。(3)运动前推拿或运动后推拿均能降低模型大鼠血清·OH和MDA的生成,提高SOD活性和SOD/MDA的比值,运动前推拿能更有效地抑制模型大鼠血清·OH和MDA的生成,表明推拿治疗具有良好的抗脂质过氧化作用。(4)本研究重点观察了运动前推拿与运动后推拿对DOMS模型大鼠在恢复期的不同时相血清中LD、LDH、CK、·OH、MDA和SOD的变化规律;研究结果表明,运动前推拿能降低血清内CK和LDH活性,运动前推拿或运动后推拿能抑制血清中·OH与MDA的生成,提高血清中SOD的活性和SOD/MDA的比值,间接说明推拿能延缓大鼠DOMS的产生,加速肌肉酸痛的消除,有效控制脂质过氧化损伤,提高机体抗氧化能力,对肌细胞具有良好的保护作用。
This dissertation includes two parts, i.e. literature research and animal experiment. Literature research
Delayed-onset muscle soreness (DOMS) describes muscle pain and tenderness that typically develop several hours postexercise and consist of predominantly eccentric muscle actions, especially if the exercise is unfamiliar. As we know, traditional Chinese Tuina is widely used in sports teams with its unique advantages in relieving exercise-induced fatigue and strengthening sporting abilities, and it has gained more and more attention in prevention and treatment of DOMS. This part reviews the relative literatures in western and traditional medicine, and sums up the pathogenesis and evaluation of DOMS, and approaches to prevention and treatment for DOMS. This part also covers the mechanism of Tuina on preventing and treating sports injuries, which provides theoretical basis for prevention and treatment on DOMS by Tuina. The design of animal experiment, observing data and experimental method were established on the basis of these literatures.
Animal experiment research
Objective: to evaluate the effects of Tuina on DOMS and the relationship of oxidative-antioxidative balance disturbance in rats by testing serum LD, LDH, CK, MDA,SOD、·OH.
Method
130 healthy and clean male SD rats of six-week old were randomly divided into 4 groups including normal control group(A), exercise control group(B), Tuina pre-exercise group(C) and Tuina post-exercise(D). Group B, C and D were randomly divided into 1h, 12h, 24h and 48h groups. Each group consists of 10 rats. Group B were put into a glass pool of 100 cm×60 cm×70 cm to swim carrying a subject of 3% its body weight till exhausted. Group C and Group D were treated 10 minutes by Tuina pre-exercise or post-exercise. The rats in Group B, C and D were killed to take blood sample 1h, 12h, 24h and 48h post-exercise or after Tuina. While the normal control group were killed after 48h to take blood sample.
Results
(1) In comparison with the normal control group, the level of serum LD in exercise control group, Tuina pre-exercise group and Tuina post-exercise rose significantly. In comparison with the exercise control group, the level of serum LD in Tuina pre-exercise group and Tuina post-exercise lowered significantly. (2) The level of serum LDH rose immediately to peak after 12h postexercise. The level of serum LDH lowered in level with LD in Group B, C and D. (3) In comparison with the normal control group, the level of serum CK in group B, group C and group D rose immediately. In comparison with group B, the level of serum CK in 1h, 12h in group C had no significant difference while it rose significantly in 1h, 12h in group D. In comparison with group C, the level of serum CK after 1h,12h and 48h in group D rose signifieantly. (4) The level of serum·OH rose immediately to peak after 1h in group B. In comparison with group B after 1h,12h, the level of serum·OH lowered signifieantly in group C and group D. The level of serum OH in 1h, 12h, and 24h had no significant difference between group C and group D. (5) The level of serum MDA rose signifieantly in group B. In comparison with group A, the level of serum MDA in 1h,12h and 24h in group C had no significant difference. While the level of serum MDA after 12h, 24h and 48h in group D had no significant difference. (6) In comparison with group A and group B, the level of serum SOD rose signifieantly in 1h, 12h, 24h and 48h in both group C and group D. (7) In comparison with group A, the ratio of SOD/MDA lowered significantly in group B. In comparison with group B, the ratio of SOD/MDA rose signifieantly in 1h, 12h, 24h and 48h in both group C and group D.
Conclusion
(1) The change of serum LD is related with exercise-induced fatigue, but it is not related with DOMS. (2) Tuina pre-exercise is effective in alleviating muscle damage caused by eccentric exercise in DOMS. (3) Tuina pre-exercise or postexercise are effective in lowering the level of serum OH and MDA, while raising the activity of serum SOD and the ratio of SOD/MDA. Tuina pre-exercise is more effective. (4) This experiment focused on Tuina pre-exercise or postexercise on rats of DOMS in testing the changes of LD、LDH、CK、·OH、MDA and SOD. The experimental result shows that Tuina pre-exercise is effective in lowering the level of serum LDH and CK. Tuina pre-exercise or postexercise are effective in lowering the level of serum·OH and MDA, while raising the activity of serum SOD and the ratio of SOD/MDA. Tuina is effective in delaying DOMS, reducing muscle soreness, controlling the damage of lipid peroxidation, increasing the ability in anti-oxidation and has good protection to muscle cell.
引文
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