基于马达蛋白及NGF探寻推拿对坐骨神经损伤大鼠轴浆运输功能的影响及机理
摘要
[目的]
周围神经损伤是临床的常见病、多发病,多以感觉、运动功能的障碍为主要症状。运用推拿治疗相关疾病,在临床上取得了良好的疗效,但其中机理尚未阐明。由于神经功能恢复的基础是轴浆运输功能的正常,因此本次研究对象选为坐骨神经夹持损伤模型大鼠,以模拟临床中周围神经的压迫损伤,在通过推拿给予定时、定性、定量的干预后,以行为学、形态学作为疗效评价的指标,从马达蛋白及神经生长因子在脊髓中表达的变化,探索推拿促进轴浆运输功能恢复的机理,为临床应用推拿治疗周围神经损伤疾病,改善患者的症状及预后提供有力科学证据。
[方法]
采用SD大鼠,进行坐骨神经夹持损伤造模,选取患侧“穴位-神经-肌肉区域”中的殷门、承山、阳陵泉三穴进行推拿治疗,从行为学角度,通过对其光热耐痛阈和斜板试验的检测,评价推拿治疗后神经和肌肉的感觉、运动恢复情况；从形态学角度,通过对其脊髓、损伤点神经、肌肉的染色观察,探寻推拿促进神经元再生修复的证据；并进而通过检测脊髓中马达蛋白的表达,评价推拿对轴浆运输功能动力支持的影响；检测脊髓中动力蛋白激活蛋白与神经生长因子的表达,找寻推拿促进轴浆运输功能恢复的证据,最终进一步理清推拿治疗坐骨神经损伤的作用机理和起效途径等。
[结果]
1.行为学
光热耐痛阈结果如下：造模7d后、推拿治疗10次、20次后,左侧(健侧)光热耐痛阈各组间均无差异；造模7d后右侧(患侧)光热耐痛阈,模型组与正常组相比均有明显差异(P<0.05),说明坐骨神经损伤后,大鼠的感觉功能减退,敏感度降低。推拿治疗10次、20次后,大鼠右侧光热耐痛阈推拿治疗组与模型组和模型对照组相比均有降低(P<0.05),痛敏分数推拿治疗组与模型组相比有显著性降低(P<0.01),说明推拿可以促进坐骨神经损伤大鼠感觉功能的恢复。
斜板试验结果表明,造模7d后模型组与正常组相比有明显差异(p<0.05),说明坐骨神经损伤后,大鼠的运动功能减退,肌力降低,造模成功。推拿治疗10次后,推拿治疗组与模型组和模型对照组相比均有显著性升高(p<0.05),且已接近正常组水平。推拿治疗20次后,推拿治疗组已达到甚至超过正常组水平,且与模型组和模型对照组相比仍有显著性差异(p<0.05),可能与推拿帮助改善肌力的功效有关。说明推拿可以有效促进坐骨神经损伤大鼠运动功能和下肢肌力的恢复。
2.形态学
腓肠肌湿重结果如下：造模7d后、推拿治疗10次、20次后,各组大鼠左侧腓肠肌湿重均没有差异。造模7d后,假手术组与模型组大鼠患侧腓肠肌湿重均有一定程度的降低,且模型组更为明显,模型组与正常组和假手术组相比,其腓肠肌湿重和腓肠肌恢复率显著性降低(P<0.05),说明假手术组单纯肌肉的剥离对肌肉产生了一定的影响。推拿治疗10次后,推拿治疗组大鼠腓肠肌湿重和腓肠肌恢复率均有升高,且与模型组和模型对照组有显著性差异(P<0.05),但各组大鼠的腓肠肌湿重和恢复率与正常组相比仍存在差异(P<0.05)。推拿治疗20次后,推拿治疗组大鼠腓肠肌湿重和恢复率仍高于模型组和模型对照组,但与正常组相比仍有一定差距(P<0.05)
造模7d后,模型组右侧腓肠肌肌细胞直径明显小于正常组和假手术组,差异具有统计学意义(P<0.05)。推拿治疗10次后,推拿治疗组大鼠右侧腓肠肌肌细胞直径有一定程度增大,且与模型组和模型对照组比较存在显著差异(P<0.05),但与正常组相比仍然较低(P<0.05)。推拿治疗20次后,各组大鼠右侧腓肠肌肌细胞直径皆逐渐升高,且推拿治疗组明显优于模型组和模型对照组(P<0.05),但尚未达到正常组水平。
脊髓HE染色结果：正常情况下,脊髓腹角神经元细胞排列整齐,细胞核位于细胞中央,核仁清晰,尼氏体均匀分布；模型组脊髓腹角神经元排列不规则,数量减少,细胞出现肿胀、变性等现象,染色较浅,胞核移向边缘,尼氏体溶解消失；经推拿治疗后,脊髓腹角神经元变性、损伤程度较轻,排列较规则,部分细胞核移向边缘,可见少量细胞核再生。
坐骨神经HE染色结果：正常情况下,神经元轴突及雪旺细胞排列整齐；模型组部分神经元轴突及雪旺细胞排列不规则,出现肿胀、破溃,甚至崩裂,而被巨噬细胞吞噬,胶质细胞出现明显增生；经推拿治疗后雪旺细胞及轴突基本正常。
腓肠肌HE染色结果：正常情况下,腓肠肌细胞大小一致,排列整齐,胞核清晰,胞浆染色均匀,肌纤维间隙适中；模型组腓肠肌肌细胞直径明显缩小,分布不均匀,胞核清晰,但数量明显增多,肌纤维间隙增大,呈现萎缩现象；经推拿治疗后腓肠肌肌细胞直径较均匀,胞核清晰,肌纤维萎缩程度较轻。
3.机理研究
正常组及假手术组大鼠马达蛋白(动力蛋白Dynein,驱动蛋白Kinesin、动力蛋白激活蛋白Dynactin)、NGF在脊髓中均有少量表达,阳性神经元的胞核呈蓝色,胞浆呈棕褐色；造模7d后,模型组大鼠以上研究指标在脊髓中的表达均较正常组和假手术组染色加深,免疫组化的平均光密度亦增高,具有统计学意义(P<0.05)；推拿治疗10次后,模型组、模型对照组及推拿治疗组大鼠脊髓中Dynein、Kinesin、Dynactin、NGF的免疫组化平均光密度均较正常组显著增高,且推拿治疗组的染色范围和程度明显优于模型组,与其余四组比较均具有统计学意义；推拿治疗20次后,在脊髓中推拿治疗组的马达蛋白及NGF的表达与其余四组比较仍有显著性差异,而Dynactin在推拿组中的表达亦维持在较高水平,但是模型组中的表达却降低到与正常组无显著性差异的水平,结合模型组大鼠的行为学表现,推断模型组大鼠的逆向轴浆运输功能发生障碍,因此即使NGF能够高表达,也无法被及时转运至神经元胞体,从而导致周围神经的损伤没有得到及时修复。考虑到受治疗时间的限制,按实验结果分析,如果继续治疗,当受损神经恢复到一定程度,Kinesin及Dynein的表达会逐渐降低,趋于正常。模型组中的表达情况也是如此,但推拿组的轴浆运输功能会得到改善。因而在最终比较时,虽然Kinesin及Dynein的表达在推拿组与模型组间比较无显著性差异,但是大鼠的行为学会出现较大差异,即推拿组要明显优于模型组。这得益于通过推拿,提高了脊髓中马达蛋白的表达,促进了轴浆运输功能的恢复。
[结论]
1推拿可以提高坐骨神经损伤大鼠腓肠肌的肌肉恢复率以及肌力,并能提升大鼠的痛觉敏感程度,从而促进坐骨神经损伤大鼠运动和感觉功能的恢复；
2推拿能够提高坐骨神经损伤大鼠脊髓中马达蛋白的表达,为轴浆运输功能的恢复提供了动力基础,为神经元与靶器官之间建立了营养传递通路；
3推拿能够提高坐骨神经损伤大鼠脊髓中动力蛋白激活蛋白与神经生长因子的表达,为神经元的存活与再生、受损轴突的出芽与生长提供营养保障。
4从马达蛋白动力层面促进轴浆运输的发生,并进而从神经营养物质传递层面改善轴浆运输的功能,应该为推拿治疗周围神经损伤的起效机理之一
[Objective]
Peripheral nerve injury is a clinical common disease, multi-sensory and motor function disorders are the main symptoms. The use of massage therapy-related diseases in clinical achieved good results, but has not been elucidated. Recovery of neurological function is based on axonal transport function in normal, selected as the object of this study sciatic nerve clamping injury rat model to simulate the clinical peripheral nerve compression injury, timing given by massage, qualitative and quantitative intervention, behavior, morphology as efficacy evaluation indicators from the motor protein and nerve growth factor expression changes in the spinal cord, discover massage promote axonal transport functional recovery mechanism, the clinical application of massage for treatment of peripheral nerve injury diseases, improve symptoms and prognosis of the patient to provide strong scientific evidence.
[Method]
SD rats, carried out the sciatic nerve clamping injury model, select the ipsilateral acupuncture points-nerve-muscle area "Yin door, Chengshan, Yanglingquan hole for massage therapy, from the behavioral point of view, by its light heat pain tolerance threshold and the detection of the inclined plane test, evaluation massage therapy feeling of nerves and muscles, exercise recovery; massage from a morphological point of view, through its spinal cord, damage points nervous, muscle staining, to explore the promotion of neuronal regeneration evidence of repair; through detection of spinal cord motor protein, evaluation and massage axonal transport function power support; detect activated protein dynein in the spinal cord and nerve growth factor expression, to find massage to promote axonal transport function recovery evidence, and ultimately to further clarify the mechanism of action of the massage treatment of sciatic nerve injury and the onset of ways.
[Results]
1.behavior
Light and heat-resistant pain threshold results are as follows:modeling7days after the massage therapy10times,20times, left (contralateral) light and heat pain tolerance threshold between the groups, there were no differences; modeling7d right side (ipsilateral) light and heat pain tolerance threshold, the model group and normal group were significantly different (P<0.05) after sciatic nerve injury in rats sensory dysfunction, reduced sensitivity. Massage therapy10times,20times, compared to the rat right light and heat pain tolerance threshold massage treatment group and model group and med(?) control group were lower-(P<0.05), the hyperalgesia fraction massage therapy group compared with the model group significantly decreased (P<0.01), massage can promote the recovery of sensory function of the sciatic nerve injury in rats.
Inclined plane test results show that the model group and normal group modeling7d significant difference (p<0.05) after sciatic nerve injury in rats motor function loss, decreased muscle strength, successful modeling.10massage therapy, massage therapy group and model group and model control group compared were significantly elevated (p<0.05), and close to the level of the normal group.20massage therapy, massage therapy group to meet or exceed the level of the normal group, and there is still a significant difference (p<0.05), massage may help improve the effectiveness of the muscle strength compared with the model group and model control group. Massage can effectively promote the recovery of the sciatic nerve injury in rats motor function and lower extremity muscle strength.
2.Morphology
Gastrocnemius wet weight results are as follows:7d modeling, massage therapy10times,20times, the rats left gastrocnemius muscle wet weight There were no differences. Modeling7d, sham-operated group and model group were ipsilateral gastrocnemius wet weight have a certain degree of reduction is more obvious, and the model group, model group and the normal group and sham operation group, the gastrocnemius muscle wet weight and gastrocnemius The recovery rate was significantly decreased (P<0.05), stripped of pure muscle of the sham-operated group had a definite impact on muscle.10massage therapy, massage therapy group rat gastrocnemius muscle wet weight and the gastrocnemius muscle recovery rate increased, and significant difference (P<0.05) with the model group and model control group, but the gastrocnemius wet weight of the rats in each group and the recovery rate and the normal group, there is still a difference (P<0.05).20massage therapy, massage therapy group gastrocnemius wet weight and the recovery rate is still higher than the model group and model control group, but still a gap compared with the normal group (P<0.05).
Model7d, the diameter of the gastrocnemius muscle cells of the right side of the model group was significantly less than the normal group and sham operation group, the difference was statistically significant (P<0.05).10massage therapy, massage therapy group the right side of the rat gastrocnemius muscle cell diameter increased to a certain extent, and there is a significant difference compared with the model group and model control group (P<0.05), but is still low compared with the normal group (P<0.05). Massage therapy for20rats in each group the right gastrocnemius muscle cell diameter are gradually increased, and massage treatment group was significantly better than the model group and model control group (P <0.05), but has not yet reached the level of the normal group.
HE staining in the spinal cord:Under normal circumstances, the ventral horn of the spinal cord neurons arranged in neat rows, the nucleus is located in the center of the cell, the nucleolus is clear, evenly distributed Nissl bodies; model of spinal cord ventral horn neurons irregular arrangement, reducing the number of cell swelling degeneration phenomenon, lighter staining nucleus toward the edge of Nissl bodies dissolved disappear; after massage therapy, spinal cord ventral horn neuronal degeneration, damage to a lesser extent, arranged in a more regular part of the nucleus toward the edge, we can see a small amount of reproducing the nucleus.
The sciatic nerve HE staining:Under normal circumstances, the axons and Schwann cells arranged in neat rows; model group, part of the axon and Schwann cells arranged in irregular, swelling, ulceration, or even crack, and macrophages glial cells significantly hyperplasia; Schwann cells and axons normal after massage therapy.
The gastrocnemius HE staining results:Under normal circumstances, the gastrocnemius muscle in the same size, arranged in neat rows, clear nucleus and cytoplasm uniform moderate muscle fiber gap; diameter of the gastrocnemius muscle cells of the model group was significantly reduced, the uneven distribution of the nucleus is clear, but the number is significantly increased, increased muscle fiber gap shrank phenomenon; uniform diameter of the gastrocnemius muscle cells after massage therapy, clear nucleus, to a lesser extent of muscle fiber atrophy.
3. Mechanism
Normal group and sham-operated rats motor protein (dynein Dynein, Kinesin Kinesin, dynein activator protein Dynactin), NGF in the spinal cord have a small amount of expression of positive neurons in the nucleus was blue, the cytoplasm tan; modeling7d, more than model group research indicators expression in the spinal cord compared with the normal group and sham operation group deeper staining, immunohistochemistry average optical density also increased, with statistical significance (P<0.05); massage10after treatment, the model group, model control group and the massage therapy group rat spinal cord Dynein, Kinesin, Dynactin, NGF immunohistochemistry average optical density compared with the normal group was significantly higher, and the the dyeing range and extent of the massage therapy group was significantly better than the model group, and the remaining four groups were statistically significant; massage therapy after20, the motor protein of the massage therapy group and the expression of NGF in the spinal cord and the remaining four groups comparison there is still a significant difference, and Dynactin massage Reverse axonal transport group expression at a higher level, but the expression in the model group was reduced to with the normal group there was no significant difference in the level, combined with the performance of the model group behavior, the inference model rats barrier function, NGF high expression, it can not be transported to the neuronal cell bodies in a timely manner, resulting in peripheral nerve damage has not been repaired in time. Taking into account the treatment time constraints, according to the experimental results, continue treatment, when damaged nerve recovery to a certain extent, the Kinesin and Dynein expression, will gradually reduce to normal. Expression in the model group, too, but the massage group of axonal transport function will be improved. Thus in the final comparison, although the Kinesin and Dynein expression, in the massage with the model group no significant difference, but the behavior of the rats learn to appear quite different, namely, the massage group was significantly better than the model group. Massage, thanks to improved spinal cord motor protein, promote axonal transport functional recovery.
[Conclusion]
1massage can improve the the gastroenemius muscle recovery rate of sciatic nerve injury in rats as well as muscle strength, and can enhance the degree of hyperalgesia in rats, thus contributing to the recovery of the sciatic nerve injury in rat motor and sensory function;
2massage can improve the sciatic nerve motor protein expression in the spinal cord injury in rats, the axis the slurry transport functional recovery provided the impetus foundation, nutrition pathway between neurons and target organ;
3massage can improve the sciatic nerve the dynein activation of protein in the spinal cord injury in rats with nerve growth factor expression, neuronal survival and regeneration, impaired axonal sprouting and growth nutrition security.
4from motor protein power level to promote axonal transport occurrence and improve axonai transport of neurotrophic substances pass level, should be one of the massage therapy onset mechanism of peripheral nerve injury.
周围神经损伤是临床的常见病、多发病,多以感觉、运动功能的障碍为主要症状。运用推拿治疗相关疾病,在临床上取得了良好的疗效,但其中机理尚未阐明。由于神经功能恢复的基础是轴浆运输功能的正常,因此本次研究对象选为坐骨神经夹持损伤模型大鼠,以模拟临床中周围神经的压迫损伤,在通过推拿给予定时、定性、定量的干预后,以行为学、形态学作为疗效评价的指标,从马达蛋白及神经生长因子在脊髓中表达的变化,探索推拿促进轴浆运输功能恢复的机理,为临床应用推拿治疗周围神经损伤疾病,改善患者的症状及预后提供有力科学证据。
[方法]
采用SD大鼠,进行坐骨神经夹持损伤造模,选取患侧“穴位-神经-肌肉区域”中的殷门、承山、阳陵泉三穴进行推拿治疗,从行为学角度,通过对其光热耐痛阈和斜板试验的检测,评价推拿治疗后神经和肌肉的感觉、运动恢复情况；从形态学角度,通过对其脊髓、损伤点神经、肌肉的染色观察,探寻推拿促进神经元再生修复的证据；并进而通过检测脊髓中马达蛋白的表达,评价推拿对轴浆运输功能动力支持的影响；检测脊髓中动力蛋白激活蛋白与神经生长因子的表达,找寻推拿促进轴浆运输功能恢复的证据,最终进一步理清推拿治疗坐骨神经损伤的作用机理和起效途径等。
[结果]
1.行为学
光热耐痛阈结果如下：造模7d后、推拿治疗10次、20次后,左侧(健侧)光热耐痛阈各组间均无差异；造模7d后右侧(患侧)光热耐痛阈,模型组与正常组相比均有明显差异(P<0.05),说明坐骨神经损伤后,大鼠的感觉功能减退,敏感度降低。推拿治疗10次、20次后,大鼠右侧光热耐痛阈推拿治疗组与模型组和模型对照组相比均有降低(P<0.05),痛敏分数推拿治疗组与模型组相比有显著性降低(P<0.01),说明推拿可以促进坐骨神经损伤大鼠感觉功能的恢复。
斜板试验结果表明,造模7d后模型组与正常组相比有明显差异(p<0.05),说明坐骨神经损伤后,大鼠的运动功能减退,肌力降低,造模成功。推拿治疗10次后,推拿治疗组与模型组和模型对照组相比均有显著性升高(p<0.05),且已接近正常组水平。推拿治疗20次后,推拿治疗组已达到甚至超过正常组水平,且与模型组和模型对照组相比仍有显著性差异(p<0.05),可能与推拿帮助改善肌力的功效有关。说明推拿可以有效促进坐骨神经损伤大鼠运动功能和下肢肌力的恢复。
2.形态学
腓肠肌湿重结果如下：造模7d后、推拿治疗10次、20次后,各组大鼠左侧腓肠肌湿重均没有差异。造模7d后,假手术组与模型组大鼠患侧腓肠肌湿重均有一定程度的降低,且模型组更为明显,模型组与正常组和假手术组相比,其腓肠肌湿重和腓肠肌恢复率显著性降低(P<0.05),说明假手术组单纯肌肉的剥离对肌肉产生了一定的影响。推拿治疗10次后,推拿治疗组大鼠腓肠肌湿重和腓肠肌恢复率均有升高,且与模型组和模型对照组有显著性差异(P<0.05),但各组大鼠的腓肠肌湿重和恢复率与正常组相比仍存在差异(P<0.05)。推拿治疗20次后,推拿治疗组大鼠腓肠肌湿重和恢复率仍高于模型组和模型对照组,但与正常组相比仍有一定差距(P<0.05)
造模7d后,模型组右侧腓肠肌肌细胞直径明显小于正常组和假手术组,差异具有统计学意义(P<0.05)。推拿治疗10次后,推拿治疗组大鼠右侧腓肠肌肌细胞直径有一定程度增大,且与模型组和模型对照组比较存在显著差异(P<0.05),但与正常组相比仍然较低(P<0.05)。推拿治疗20次后,各组大鼠右侧腓肠肌肌细胞直径皆逐渐升高,且推拿治疗组明显优于模型组和模型对照组(P<0.05),但尚未达到正常组水平。
脊髓HE染色结果：正常情况下,脊髓腹角神经元细胞排列整齐,细胞核位于细胞中央,核仁清晰,尼氏体均匀分布；模型组脊髓腹角神经元排列不规则,数量减少,细胞出现肿胀、变性等现象,染色较浅,胞核移向边缘,尼氏体溶解消失；经推拿治疗后,脊髓腹角神经元变性、损伤程度较轻,排列较规则,部分细胞核移向边缘,可见少量细胞核再生。
坐骨神经HE染色结果：正常情况下,神经元轴突及雪旺细胞排列整齐；模型组部分神经元轴突及雪旺细胞排列不规则,出现肿胀、破溃,甚至崩裂,而被巨噬细胞吞噬,胶质细胞出现明显增生；经推拿治疗后雪旺细胞及轴突基本正常。
腓肠肌HE染色结果：正常情况下,腓肠肌细胞大小一致,排列整齐,胞核清晰,胞浆染色均匀,肌纤维间隙适中；模型组腓肠肌肌细胞直径明显缩小,分布不均匀,胞核清晰,但数量明显增多,肌纤维间隙增大,呈现萎缩现象；经推拿治疗后腓肠肌肌细胞直径较均匀,胞核清晰,肌纤维萎缩程度较轻。
3.机理研究
正常组及假手术组大鼠马达蛋白(动力蛋白Dynein,驱动蛋白Kinesin、动力蛋白激活蛋白Dynactin)、NGF在脊髓中均有少量表达,阳性神经元的胞核呈蓝色,胞浆呈棕褐色；造模7d后,模型组大鼠以上研究指标在脊髓中的表达均较正常组和假手术组染色加深,免疫组化的平均光密度亦增高,具有统计学意义(P<0.05)；推拿治疗10次后,模型组、模型对照组及推拿治疗组大鼠脊髓中Dynein、Kinesin、Dynactin、NGF的免疫组化平均光密度均较正常组显著增高,且推拿治疗组的染色范围和程度明显优于模型组,与其余四组比较均具有统计学意义；推拿治疗20次后,在脊髓中推拿治疗组的马达蛋白及NGF的表达与其余四组比较仍有显著性差异,而Dynactin在推拿组中的表达亦维持在较高水平,但是模型组中的表达却降低到与正常组无显著性差异的水平,结合模型组大鼠的行为学表现,推断模型组大鼠的逆向轴浆运输功能发生障碍,因此即使NGF能够高表达,也无法被及时转运至神经元胞体,从而导致周围神经的损伤没有得到及时修复。考虑到受治疗时间的限制,按实验结果分析,如果继续治疗,当受损神经恢复到一定程度,Kinesin及Dynein的表达会逐渐降低,趋于正常。模型组中的表达情况也是如此,但推拿组的轴浆运输功能会得到改善。因而在最终比较时,虽然Kinesin及Dynein的表达在推拿组与模型组间比较无显著性差异,但是大鼠的行为学会出现较大差异,即推拿组要明显优于模型组。这得益于通过推拿,提高了脊髓中马达蛋白的表达,促进了轴浆运输功能的恢复。
[结论]
1推拿可以提高坐骨神经损伤大鼠腓肠肌的肌肉恢复率以及肌力,并能提升大鼠的痛觉敏感程度,从而促进坐骨神经损伤大鼠运动和感觉功能的恢复；
2推拿能够提高坐骨神经损伤大鼠脊髓中马达蛋白的表达,为轴浆运输功能的恢复提供了动力基础,为神经元与靶器官之间建立了营养传递通路；
3推拿能够提高坐骨神经损伤大鼠脊髓中动力蛋白激活蛋白与神经生长因子的表达,为神经元的存活与再生、受损轴突的出芽与生长提供营养保障。
4从马达蛋白动力层面促进轴浆运输的发生,并进而从神经营养物质传递层面改善轴浆运输的功能,应该为推拿治疗周围神经损伤的起效机理之一
[Objective]
Peripheral nerve injury is a clinical common disease, multi-sensory and motor function disorders are the main symptoms. The use of massage therapy-related diseases in clinical achieved good results, but has not been elucidated. Recovery of neurological function is based on axonal transport function in normal, selected as the object of this study sciatic nerve clamping injury rat model to simulate the clinical peripheral nerve compression injury, timing given by massage, qualitative and quantitative intervention, behavior, morphology as efficacy evaluation indicators from the motor protein and nerve growth factor expression changes in the spinal cord, discover massage promote axonal transport functional recovery mechanism, the clinical application of massage for treatment of peripheral nerve injury diseases, improve symptoms and prognosis of the patient to provide strong scientific evidence.
[Method]
SD rats, carried out the sciatic nerve clamping injury model, select the ipsilateral acupuncture points-nerve-muscle area "Yin door, Chengshan, Yanglingquan hole for massage therapy, from the behavioral point of view, by its light heat pain tolerance threshold and the detection of the inclined plane test, evaluation massage therapy feeling of nerves and muscles, exercise recovery; massage from a morphological point of view, through its spinal cord, damage points nervous, muscle staining, to explore the promotion of neuronal regeneration evidence of repair; through detection of spinal cord motor protein, evaluation and massage axonal transport function power support; detect activated protein dynein in the spinal cord and nerve growth factor expression, to find massage to promote axonal transport function recovery evidence, and ultimately to further clarify the mechanism of action of the massage treatment of sciatic nerve injury and the onset of ways.
[Results]
1.behavior
Light and heat-resistant pain threshold results are as follows:modeling7days after the massage therapy10times,20times, left (contralateral) light and heat pain tolerance threshold between the groups, there were no differences; modeling7d right side (ipsilateral) light and heat pain tolerance threshold, the model group and normal group were significantly different (P<0.05) after sciatic nerve injury in rats sensory dysfunction, reduced sensitivity. Massage therapy10times,20times, compared to the rat right light and heat pain tolerance threshold massage treatment group and model group and med(?) control group were lower-(P<0.05), the hyperalgesia fraction massage therapy group compared with the model group significantly decreased (P<0.01), massage can promote the recovery of sensory function of the sciatic nerve injury in rats.
Inclined plane test results show that the model group and normal group modeling7d significant difference (p<0.05) after sciatic nerve injury in rats motor function loss, decreased muscle strength, successful modeling.10massage therapy, massage therapy group and model group and model control group compared were significantly elevated (p<0.05), and close to the level of the normal group.20massage therapy, massage therapy group to meet or exceed the level of the normal group, and there is still a significant difference (p<0.05), massage may help improve the effectiveness of the muscle strength compared with the model group and model control group. Massage can effectively promote the recovery of the sciatic nerve injury in rats motor function and lower extremity muscle strength.
2.Morphology
Gastrocnemius wet weight results are as follows:7d modeling, massage therapy10times,20times, the rats left gastrocnemius muscle wet weight There were no differences. Modeling7d, sham-operated group and model group were ipsilateral gastrocnemius wet weight have a certain degree of reduction is more obvious, and the model group, model group and the normal group and sham operation group, the gastrocnemius muscle wet weight and gastrocnemius The recovery rate was significantly decreased (P<0.05), stripped of pure muscle of the sham-operated group had a definite impact on muscle.10massage therapy, massage therapy group rat gastrocnemius muscle wet weight and the gastrocnemius muscle recovery rate increased, and significant difference (P<0.05) with the model group and model control group, but the gastrocnemius wet weight of the rats in each group and the recovery rate and the normal group, there is still a difference (P<0.05).20massage therapy, massage therapy group gastrocnemius wet weight and the recovery rate is still higher than the model group and model control group, but still a gap compared with the normal group (P<0.05).
Model7d, the diameter of the gastrocnemius muscle cells of the right side of the model group was significantly less than the normal group and sham operation group, the difference was statistically significant (P<0.05).10massage therapy, massage therapy group the right side of the rat gastrocnemius muscle cell diameter increased to a certain extent, and there is a significant difference compared with the model group and model control group (P<0.05), but is still low compared with the normal group (P<0.05). Massage therapy for20rats in each group the right gastrocnemius muscle cell diameter are gradually increased, and massage treatment group was significantly better than the model group and model control group (P <0.05), but has not yet reached the level of the normal group.
HE staining in the spinal cord:Under normal circumstances, the ventral horn of the spinal cord neurons arranged in neat rows, the nucleus is located in the center of the cell, the nucleolus is clear, evenly distributed Nissl bodies; model of spinal cord ventral horn neurons irregular arrangement, reducing the number of cell swelling degeneration phenomenon, lighter staining nucleus toward the edge of Nissl bodies dissolved disappear; after massage therapy, spinal cord ventral horn neuronal degeneration, damage to a lesser extent, arranged in a more regular part of the nucleus toward the edge, we can see a small amount of reproducing the nucleus.
The sciatic nerve HE staining:Under normal circumstances, the axons and Schwann cells arranged in neat rows; model group, part of the axon and Schwann cells arranged in irregular, swelling, ulceration, or even crack, and macrophages glial cells significantly hyperplasia; Schwann cells and axons normal after massage therapy.
The gastrocnemius HE staining results:Under normal circumstances, the gastrocnemius muscle in the same size, arranged in neat rows, clear nucleus and cytoplasm uniform moderate muscle fiber gap; diameter of the gastrocnemius muscle cells of the model group was significantly reduced, the uneven distribution of the nucleus is clear, but the number is significantly increased, increased muscle fiber gap shrank phenomenon; uniform diameter of the gastrocnemius muscle cells after massage therapy, clear nucleus, to a lesser extent of muscle fiber atrophy.
3. Mechanism
Normal group and sham-operated rats motor protein (dynein Dynein, Kinesin Kinesin, dynein activator protein Dynactin), NGF in the spinal cord have a small amount of expression of positive neurons in the nucleus was blue, the cytoplasm tan; modeling7d, more than model group research indicators expression in the spinal cord compared with the normal group and sham operation group deeper staining, immunohistochemistry average optical density also increased, with statistical significance (P<0.05); massage10after treatment, the model group, model control group and the massage therapy group rat spinal cord Dynein, Kinesin, Dynactin, NGF immunohistochemistry average optical density compared with the normal group was significantly higher, and the the dyeing range and extent of the massage therapy group was significantly better than the model group, and the remaining four groups were statistically significant; massage therapy after20, the motor protein of the massage therapy group and the expression of NGF in the spinal cord and the remaining four groups comparison there is still a significant difference, and Dynactin massage Reverse axonal transport group expression at a higher level, but the expression in the model group was reduced to with the normal group there was no significant difference in the level, combined with the performance of the model group behavior, the inference model rats barrier function, NGF high expression, it can not be transported to the neuronal cell bodies in a timely manner, resulting in peripheral nerve damage has not been repaired in time. Taking into account the treatment time constraints, according to the experimental results, continue treatment, when damaged nerve recovery to a certain extent, the Kinesin and Dynein expression, will gradually reduce to normal. Expression in the model group, too, but the massage group of axonal transport function will be improved. Thus in the final comparison, although the Kinesin and Dynein expression, in the massage with the model group no significant difference, but the behavior of the rats learn to appear quite different, namely, the massage group was significantly better than the model group. Massage, thanks to improved spinal cord motor protein, promote axonal transport functional recovery.
[Conclusion]
1massage can improve the the gastroenemius muscle recovery rate of sciatic nerve injury in rats as well as muscle strength, and can enhance the degree of hyperalgesia in rats, thus contributing to the recovery of the sciatic nerve injury in rat motor and sensory function;
2massage can improve the sciatic nerve motor protein expression in the spinal cord injury in rats, the axis the slurry transport functional recovery provided the impetus foundation, nutrition pathway between neurons and target organ;
3massage can improve the sciatic nerve the dynein activation of protein in the spinal cord injury in rats with nerve growth factor expression, neuronal survival and regeneration, impaired axonal sprouting and growth nutrition security.
4from motor protein power level to promote axonal transport occurrence and improve axonai transport of neurotrophic substances pass level, should be one of the massage therapy onset mechanism of peripheral nerve injury.
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