推拿对大鼠坐骨神经损伤后NTFs的影响

【作者】 袁之琳；

【导师】 于天源；

【作者基本信息】 北京中医药大学 ， 针灸推拿学， 2011， 硕士

【摘要】 [目的]基于推拿对周围神经损伤有着良好的临床疗效,从实验研究角度探索推拿对周围神经损伤的修复机制,通过分析坐骨神经损伤大鼠的行为学、形态学及神经生长因子(NGF)、睫状神经营养因子(CNTF)、脑源性神经营养因子(BDNF)、碱性成纤维细胞生长因子(bFGF)来研究推拿对周围神经的治疗机理。[方法]本课题基于以往的实验,选用SD大鼠作为实验动物,采用坐骨神经夹持损伤模型,使用“推拿手法模拟器”依次刺激大鼠右侧(手术侧)殷门、承山、阳陵泉,施行点法→拨法→揉法进行治疗。通过斜板试验、热痛阈、腓肠肌湿重的分析,判断推拿对周围神经损伤后肢体障碍的恢复作用；通过光镜观察腓肠肌细胞HE染色,探寻推拿对失神经肌肉萎缩的影响；通过光镜观察坐骨神经HE染色,探寻推拿对神经元的保护作用；通过对肌肉、血浆的Elisa定量分析、免疫组化分析,分析神经生长因子(NGF)、脑源性神经营养因子(BDNF)、睫状神经营养因子(CNTF)、碱性成纤维细胞生长因子(bFGF)的变化,探寻推拿对相关因子表达的影响。[结果]1斜板试验由实验结果显示推拿治疗组经过推拿的治疗与模型组是有明显的差异的。证实推拿对大鼠周围神经损伤的肌力恢复有效果。2耐痛阈经过10天治疗后,推拿组与模型组有显著差异。证实推拿治疗对大鼠的热感觉恢复有显著效果。3腓肠肌湿重大鼠腓肠肌湿重明显下降。经治疗,模型组与推拿治疗组的腓肠肌湿重及其恢复率无差异。4腓肠肌细胞HE染色正常组显示肌细胞饱满、肌细胞分布均匀、肌细胞核分布均匀、肌纤维间隙正常。模型组显现肌纤维间隙变大、肌细胞萎缩直径减小、胞核增多。模型对照组显示肌纤维间隙变大但程度比模型组轻,肌细胞萎缩直径减少、胞核增多。推拿治疗组显示肌纤维间隙变大但程度要比模型组、模型对照组轻,肌细胞萎缩直径减少。5坐骨神经HE染色正常组显示轴突及雪旺细胞排列整齐。模型组显示有些轴突肿胀、轴突及雪旺细胞破溃、排列杂乱,胶质细胞明显增生。模型对照组同模型组。推拿治疗组基本同正常组。6 NGF、bFGF、BDNF、CNTF Elisa定量分析从Elisa结果来看,NGF、CNTF、BDNF腓肠肌肌肉中的含量高于血浆中含量,bFGF腓肠肌肌肉中的含量低于血浆中含量。NGF治疗后血浆组间对比无差异P>0.05。治疗后腓肠肌肌肉组间对比：正常组、模型对照组、推拿治疗组与模型组对比有显著差异P<0.05。CNTF治疗后血浆组间对比：推拿治疗组与模型对照组有显著差异P<0.05。治疗后腓肠肌肌肉组间对比：模型组与正常组、模型对照组有显著差异P<0.05。BDNF治疗后血浆组间对比：正常组、模型对照组、推拿治疗组与模型组对比有显著差异P<0.05。治疗后腓肠肌肌肉组间对比：正常组、模型对照组、推拿治疗组与模型组对比有显著差异P<0.05。bFGF治疗后血浆组间对比：模型组、模型对照组与正常组有显著差异P<0.05,模型组、模型对照组与推拿组有显著差异P<0.05。治疗后腓肠肌肌肉组间对比：模型组、模型对照组与正常组有显著差异P<0.05。7免疫组化分析免疫组化光密度分析NGF、bFGF、BDNF、CNTF在推拿组均高表达。NGF治疗后模型对照组与正常组、模型组、推拿治疗组有显著差异P<0.05。CNTF治疗后推拿治疗组与正常组、模型组、模型对照组有显著差异P<0.05。BDNF治疗后正常组、模型对照组与推拿治疗组有显著差异P<0.05。bFGF治疗后正常组、模型对照组与推拿治疗组有显著差异P<0.05。[结论]1推拿对大鼠的肌力恢复、热痛觉恢复有促进作用。2说明推拿对坐骨神经损伤后大鼠的运动神经元具有一定的保护作用。3推拿治疗10天后,因时间过短所以对肌肉萎缩产生的作用并未显现。4大鼠坐骨神经损伤后通过推拿治疗NGF、bFGF、BDNF、CNTF可以有效地促进神经血管修复、保护神经元、重建神经轴浆逆向运输的通路。更多还原

【Abstract】 [Objective]This study is based on the good clinical effect of tuina in peripheral nerve injuries, via experiments to explore the mechanism of tuina treatment in peripheral nerve injuries. Through analyzing the behavior and morphology of rats, analyzing nerve growth factor(NGF),ciliary neurotrophic factor(CNTF), braind-deried neurotrophic factor(BDNF), basic fibroblast growth factor(bFGF), to research the mechanism of tuina treatment in peripheral nerve injuries.[Method]This topic is based on past experiments, SD rats are choosed as the experimental animal, using the model of sciatic nerve with crush injury, using "tuina technique simulator" to stimulate rat’s right side(surgical side) yinmen chengshan and yanglingquan, which executes pointing poking and keading technique. Through analyzing the inclined plane test and pain threshold and gastrocnemius weight, to explore the recovery effect of body obstacles; observing gastrocnemius HE dyed by microscope, to explore tuina effects of muscular dystrophy; observing sciatic nerve HE dyed by microscope, to explore tuina’s prtective effects of neuron; by analyzing Elisa test in gastrocnemius and in blood, and by analyzing immunohistochemistry in dorsal horn, to find NGF CNTF BDNF bFGF changes, in order to tuina affective in those neurotrophin.[Results]1 Inclined Plane TestResult shows that tuina therapy group is differ from model group. It verifies that tuina therapy is effective for recovering muscle power after rat’s sciatic injury.2 Pain ThresholdResult shows that tuina therapy group is differ from model group. It verifies that tuina treatment is effective for rat’s sensorimotor recovery.3 Gastrocnemius WeightsRat’s gastrocnemius weights lose obviously. There are no differences between Tuina therapy group and model group.4 Gastrocnemius HE dyedThe normal group shows that muscle cells are full, muscle cell and nucleus distributed homogeneously, fibromuscular gap is normal.The Model group shows that fibromuscular gap is bigger and muscle cells atrophied Nuclei is increased. The Model control group shows that fibromuscular gap is bigger but less than model group, and muscle cells atrophied Nuclei is increased.The Tuina therapy group shows that fibromuscular gap is bigger but less than model group and model control group, muscle cells atrophied.5 Sciatic nerve HE dyedThe normal group shows that Axon and schwann cells arranged formally.The model group shows that axon is swelling, Axon and schwann cells bursted which arranged mess, Glial cells is increased obviously.The Model control group is same to the model group.The tuina therapy group is same to the normal group.6 NGF,CNTF,BDNF,bFGF Elisa TestThe results show that NGF CNTF BDNF contents in gastrocnemius are higher than in bloods, bFGF content in gastrocnemius is lower than in bloods.NGF result shows that comparing between plasma has no differences P>0.05. comparing between Gastrocnemius:model group has differences with others P<0.05.CNTF result shows that comparing between plasma:tuina therapy group with model control group have differences P<0.05. comparing between Gastrocnemius:model group with normal group and model control group have differences P<0.05.BDNF result shows that comparing between plasma:normal group model control group and tuina therapy group with model group have differences P<0.05. comparing between Gastrocnemius:normal group model control group and tuina group with model group have differences P<0.05.bFGF result shows that comparing between plasma:model group and model control group with normal group have differences P<0.05. comparing between Gastrocnemius:model group and model control group with normal group have differences P<0.05.7 Immunohistochemistry(IHC)IHC result shows that NGF、bFGF、BDNF、CNTF are high expression in the tuina therapy group.NGF comparing between groups show model control group with normal group model group and tuina treatment group have differences P<0.05.CNTF comparing between groups show tuina treatment group with normal group model group and model collector group have differences P<0.05.BDNF comparing between groups show normal group model group and model control group with tuina treatment group have differences P<0.05. bFGF comparing between groups show normal group model group and model collector group with tuina treatment group have differences P<0.05.[Conclusion]1 After sciatic nerve injuries, tuina treatment is effective in rats muscle power and pain feeling recovery.2 Nerves cell in sciatic in Tuina therapy group recoveries good, it tells tuina treatment is effective for sciatic nerve injuries.3 According to the results of Gastrocnemius weight shows that tuina treatment not effective yet for muscular dystrophy in 10 times treatments.4 Tuina treatment affects NGF、bFGF、BDNF、CNTF to recovery nerve injuries,it protects neuron, and it rebuilts nerve axoplasmic transportation pathway.更多还原