微波对实验性大鼠周围神经损伤后功能恢复的影响
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摘要
目的:周围神经损伤后的治疗和康复,长期以来都是医学界一个棘手的难题。近些年来医学界都在积极探索促进周围神经再生进而加速功能恢复的治疗方法。物理因子疗法广泛而有效的治疗与康复作用,早已为多年的临床实践所证实,广大患者也因此而受益。微波作为物理因子家族中的重要成员,其治疗作用的报导越来越多,临床上应用也越来越广泛,但微波对周围神经损伤后功能恢复影响的研究较少报导。本研究拟采用周围神经损伤后进行微波治疗,通过对神经生长因子(NGF)蛋白表达的影响配合大体观察、坐骨神经功能指数、腓肠肌湿重恢复率检测,探讨微波促进周围神经损伤再生的作用及作用机制,为临床治疗提供实验依据。
方法:符合实验标准健康清洁SD大鼠64只,均为成年雄性,体重250～300g,鼠龄3个月,均为雄性。应用随机数字表将动物均分为微波组与对照组,每组32只。每组再按时间段分为3天、7天、14天、28天四个时间点,共分成8个组,每组8只。应用坐骨神经钳夹术,造成重度钳夹损伤模型。术后24小时微波组的造模侧神经损伤处给予微波治疗,治疗时输出功率6W,辐射器距离造模侧大腿皮肤约3cm(相当于辐射部位的功率密度156mW/cm2),辐射持续6分钟,每天治疗一次,每周连续治疗五天,休息两天。对照组相应部位未给予任何治疗(方法同上但无输出)。于治疗后3天、7天、14天、28天进行大体观察、测量坐骨神经功能指数、取腓肠肌称其湿重计算恢复率,再取紧邻损伤部位远端坐骨神经干1cm,石蜡包埋横切片,应用免疫组织化学方法和Imagepro-plus 4.5生物医学图像分析软件,观察神经组织中神经生长因子(NGF)的表达并进行定量分析。
结果:1、大体观察:微波组4个时间段伤口愈合、步行恢复等方面疗效明显优于对照组。2、坐骨神经功能指数:3天、7天、14天、28天微波组与对照组同期比较坐骨神经功能指数提高,组间比较均有明显差异(P<0.05,P<0.01)。3、腓肠肌湿重恢复率:术后14天各组动物均开始出现了不同程度的肌肉萎缩,微波治疗组较对照组肌肉萎缩程度轻,3天、7天微波组与对照组比较无明显差异(P>0.05);14天、28天微波组与对照组比较有显著差异(P<0.05,P<0.01);4、神经生长因子(NGF)蛋白表达:治疗后3天、7天,14天、28天各组均有NGF蛋白表达,而微波治疗组出现强表达,与治疗时间相同的对照组之间存在显著差异,(P﹤0.01)。
结论:1、微波治疗能够促进大鼠周围神经损伤后神经生长因子(NGF)蛋白的表达。2、周围神经损伤后早期开始按疗程应用微波治疗,可加速周围神经功能恢复,防止肌肉萎缩发生。实验充分说明了微波具有促进神经修复的作用,为微波在临床工作中的进一步推广、应用提供实验依据。
Objective: The treatment and rehabilitation medicine of the damaged peripheral nerve has been a thorny problem for long. In recent years, the medical profession are actively exploring ways to promote regeneration and to accelerate function recovery of peripheral nerve. Physical factors therapy has been already confirmed that it has broad and effective function of treatment and rehabilitation by clinical practice for years. The majority of patients have been also benefit. Microwave ,as the important member of physical factor family, has been reported more and more of the treatment effect and has been used more and more extensive in clinic, but the study on function recovery that the microwave acted on the peripheral injury nerve has been less reported. This research used microwave to treat peripheral injured nerve. Based on the nerve growth factor (NGF) protein expression together with general observation, the influence of sciatic nerve function index recovery rate, gastrocnemius wet weight recovery rate, to discuss the effects of the microwave treatment for peripheral nerve injury, and to provide the basis for clinical treatment.
Methods: The 64 experimental standard healthy and clean SD rats were 3 months old male rats, weighing 250 ~ 300g each. They were divided into the microwave group and the control group by Random number table. every group is 32 rats . Each group were divided into four time periods (3 days, 7 days, 14 days, 28 days after treatment) by different sampling time. There were 8 groups in all and each group is 8 rats. Using a hemostat to clamp the rats’right sciatic nerve to produce severe peripheral nerve injury model .Microwave treatment was given to the injury nerve after 24 hours. Out put power of the experiment is 6W .Radiator was 3cm above the right thigh skin (equivalent to 156mW/cm of the power density of radiation), and lasted for 6 minutes. The treatment was taken once a day, and lasted for five days every week. No treatment was given to the corresponding parts of the control group. we observed the part that close to the injured distal sciatic nerve 1 km away, and tested sciatic functional index recovery rate, recovery rate of the calf gastrocnemius wet weight determination after treatment in 3,7,14,28days. Immunohistochemical experiments and Image-proplus 6.0 biomedical image analysis software would be taken to analysis neural tissue nerve growth factor (NGF) expression .In three days after treatment, 7 days, days 14 and 28 days for observation, measuring largely sciatic nerve function index, take gastrocnemius says it wet weight calculation recovery rate, take again close to injury parts distal sciatic nerve-trunk 1cm, paraffin embedding crosscut slice, applied immunohistochemical method and Imagepro - plus 4.5 biomedical image analysis software method in the study of nerve tissue nerve growth factor (NGF) expression and quantitative analysis.
Results: 1,Gross observation: The effect of the healing and walking recovery of the microwave group is significantly better than the control group. 2, The sciatic nerve function index recovery rate: The microwave group compared with the control group in 3 days, 7 days, 14 days and 28 days increased, and the difference are obvious (P < 0.05, P < 0.01). 3, Gastrocnemius wet weight recovery rate: All the animals in each group began to appear different degrees of muscle atrophy 14 days after operation, but the degree of the microwave treatment group is lighter than the control group. There is no significant difference between microwave group and the control group in both 3 days and 7 days group (P> 0.05). There is significant difference between microwave group and the control group in both 14 days and 28 days group (P< 0.05、P< 0.01).4, Nerve growth factor (NGF) protein expression: NGF protein are all expressed in each group 3 days, 7 days, 14 days, 28 days after the treatment, while it was strongly expressed in the microwave treatment group .There was significant difference ,which was statistically significant (P <0.01), between the treatment group and the control group in the same curing time period.
Conclusion: 1, microwave treatment can promote nerve growth factor (NGF) protein expression of peripheral nerve injury rat. 2, If the microwave treatment can be given early after injury, peripheral nerve function recovery will be accelerated and muscle atrophy can be prevented. The research fully illustrated that microwave can promote the recovery of the nerve. It will apply evidence for the microwave’s promotion and application in the clinical work.
方法:符合实验标准健康清洁SD大鼠64只,均为成年雄性,体重250～300g,鼠龄3个月,均为雄性。应用随机数字表将动物均分为微波组与对照组,每组32只。每组再按时间段分为3天、7天、14天、28天四个时间点,共分成8个组,每组8只。应用坐骨神经钳夹术,造成重度钳夹损伤模型。术后24小时微波组的造模侧神经损伤处给予微波治疗,治疗时输出功率6W,辐射器距离造模侧大腿皮肤约3cm(相当于辐射部位的功率密度156mW/cm2),辐射持续6分钟,每天治疗一次,每周连续治疗五天,休息两天。对照组相应部位未给予任何治疗(方法同上但无输出)。于治疗后3天、7天、14天、28天进行大体观察、测量坐骨神经功能指数、取腓肠肌称其湿重计算恢复率,再取紧邻损伤部位远端坐骨神经干1cm,石蜡包埋横切片,应用免疫组织化学方法和Imagepro-plus 4.5生物医学图像分析软件,观察神经组织中神经生长因子(NGF)的表达并进行定量分析。
结果:1、大体观察:微波组4个时间段伤口愈合、步行恢复等方面疗效明显优于对照组。2、坐骨神经功能指数:3天、7天、14天、28天微波组与对照组同期比较坐骨神经功能指数提高,组间比较均有明显差异(P<0.05,P<0.01)。3、腓肠肌湿重恢复率:术后14天各组动物均开始出现了不同程度的肌肉萎缩,微波治疗组较对照组肌肉萎缩程度轻,3天、7天微波组与对照组比较无明显差异(P>0.05);14天、28天微波组与对照组比较有显著差异(P<0.05,P<0.01);4、神经生长因子(NGF)蛋白表达:治疗后3天、7天,14天、28天各组均有NGF蛋白表达,而微波治疗组出现强表达,与治疗时间相同的对照组之间存在显著差异,(P﹤0.01)。
结论:1、微波治疗能够促进大鼠周围神经损伤后神经生长因子(NGF)蛋白的表达。2、周围神经损伤后早期开始按疗程应用微波治疗,可加速周围神经功能恢复,防止肌肉萎缩发生。实验充分说明了微波具有促进神经修复的作用,为微波在临床工作中的进一步推广、应用提供实验依据。
Objective: The treatment and rehabilitation medicine of the damaged peripheral nerve has been a thorny problem for long. In recent years, the medical profession are actively exploring ways to promote regeneration and to accelerate function recovery of peripheral nerve. Physical factors therapy has been already confirmed that it has broad and effective function of treatment and rehabilitation by clinical practice for years. The majority of patients have been also benefit. Microwave ,as the important member of physical factor family, has been reported more and more of the treatment effect and has been used more and more extensive in clinic, but the study on function recovery that the microwave acted on the peripheral injury nerve has been less reported. This research used microwave to treat peripheral injured nerve. Based on the nerve growth factor (NGF) protein expression together with general observation, the influence of sciatic nerve function index recovery rate, gastrocnemius wet weight recovery rate, to discuss the effects of the microwave treatment for peripheral nerve injury, and to provide the basis for clinical treatment.
Methods: The 64 experimental standard healthy and clean SD rats were 3 months old male rats, weighing 250 ~ 300g each. They were divided into the microwave group and the control group by Random number table. every group is 32 rats . Each group were divided into four time periods (3 days, 7 days, 14 days, 28 days after treatment) by different sampling time. There were 8 groups in all and each group is 8 rats. Using a hemostat to clamp the rats’right sciatic nerve to produce severe peripheral nerve injury model .Microwave treatment was given to the injury nerve after 24 hours. Out put power of the experiment is 6W .Radiator was 3cm above the right thigh skin (equivalent to 156mW/cm of the power density of radiation), and lasted for 6 minutes. The treatment was taken once a day, and lasted for five days every week. No treatment was given to the corresponding parts of the control group. we observed the part that close to the injured distal sciatic nerve 1 km away, and tested sciatic functional index recovery rate, recovery rate of the calf gastrocnemius wet weight determination after treatment in 3,7,14,28days. Immunohistochemical experiments and Image-proplus 6.0 biomedical image analysis software would be taken to analysis neural tissue nerve growth factor (NGF) expression .In three days after treatment, 7 days, days 14 and 28 days for observation, measuring largely sciatic nerve function index, take gastrocnemius says it wet weight calculation recovery rate, take again close to injury parts distal sciatic nerve-trunk 1cm, paraffin embedding crosscut slice, applied immunohistochemical method and Imagepro - plus 4.5 biomedical image analysis software method in the study of nerve tissue nerve growth factor (NGF) expression and quantitative analysis.
Results: 1,Gross observation: The effect of the healing and walking recovery of the microwave group is significantly better than the control group. 2, The sciatic nerve function index recovery rate: The microwave group compared with the control group in 3 days, 7 days, 14 days and 28 days increased, and the difference are obvious (P < 0.05, P < 0.01). 3, Gastrocnemius wet weight recovery rate: All the animals in each group began to appear different degrees of muscle atrophy 14 days after operation, but the degree of the microwave treatment group is lighter than the control group. There is no significant difference between microwave group and the control group in both 3 days and 7 days group (P> 0.05). There is significant difference between microwave group and the control group in both 14 days and 28 days group (P< 0.05、P< 0.01).4, Nerve growth factor (NGF) protein expression: NGF protein are all expressed in each group 3 days, 7 days, 14 days, 28 days after the treatment, while it was strongly expressed in the microwave treatment group .There was significant difference ,which was statistically significant (P <0.01), between the treatment group and the control group in the same curing time period.
Conclusion: 1, microwave treatment can promote nerve growth factor (NGF) protein expression of peripheral nerve injury rat. 2, If the microwave treatment can be given early after injury, peripheral nerve function recovery will be accelerated and muscle atrophy can be prevented. The research fully illustrated that microwave can promote the recovery of the nerve. It will apply evidence for the microwave’s promotion and application in the clinical work.
引文
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