不同针刺参数对实验性周围性面瘫兔面神经损伤再生修复影响的研究
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摘要
目的:分析各因素对疗效影响的差异,优选周围性面瘫的最佳电针治疗方案,探讨不同针刺参数对面神经损伤再生修复的机理,为提高针灸治疗本病的临床疗效提供科学实验依据。
方法:1、选用日本大耳白兔66只,雌雄各半,采用神经卡压法造成左侧面神经损伤,作为研究实验性周围性面瘫的动物模型。2、66只兔随机分为11组(每组6只),其中一组作为假手术组,一组作为模型对照组,其余九组运用正交试验设计的方法,采用针刺时机(急性期、静止期、恢复期)、电针波形(密波、疏波、疏密波)、腧穴配伍(面部+头穴、面部+体穴、面部+头体穴)的三因素三水平搭配组合方案,随机分入L_9(3~4)正交表安排的九组实验中,进行实验观察,共观察四周。3、采用电生理的方法测定面运动神经传导速度(MCV);用免疫组织化学方法对面神经核神经生长因子(NGF)和脑源性神经营养因子(BDNF)阳性神经元进行染色,并用表达阳性细胞数定量分析;用原位杂交方法对面神经核脑源性神经营养因子(BDNF mRNA)阳性神经元进行染色,并用表达阳性细胞数定量分析。4、统计学分析在SAS for windows 8.0统计软件上完成。正交分析采用直观分析及方差分析,各组间、组内比较采用方差分析、q检验和t检验。
结果:1、造模后兔出现鼻唇沟偏向右侧,胡须不能自主运动,并向后倾斜等症状,且MCV小于10m/s,比造模前明显减慢(P<0.01)。2、各组(除假手术组)造模前MCV比较,无显著性差异(P>0.05),各组(除假手术组)造模后MCV比较,无显著性差异(P>0.05),各组间具有可比性。3、经四周治疗后,电针组的MCV恢复明显快于模型组(P<0.01)。电针组面神经核内NGF蛋白、BDNF蛋白、BDNF mRNA表达水平明显高于模型组(P<0.01)。4、采用急性期、选择电针疏密波形、取面部+体穴针刺治疗实验性周围性面瘫可以使MCV恢复最快,且各因素
Objective:To analyze the extent of curative effect influenced by all factors, optimize the therapeutic scheme of electroaccupuncture peripheral facial palsy and to discuss the influence of regeneration and restoration on facial nerve injury through different stimulating parameters of acupuncture which can provide the scientific experimental evidence for improving the clinical curative effect of peripheral facial palsy treated by acupuncture.Methods:1、 We made the animal model of experimental peripheral facial palsy with sixty-six rabbits of Japan species, male and female each half, by the method of crashing nerve. 2、 Sixty-six rabbits were divided eleven groups of which each group include six rabbits randomly. Of them one group was choused as control group, another as model group, others were divided nine groups experiment arranged by L9(3~4)orthogonal table for experimental observation for four weeks through combination scheme arranged in groups by three factors and three level , consisting of the time of acupuncture which included acute period, quiescence period and convalescence, the wave shape of electroaccupuncture which included dense wave, sparse wave and density wave, the compatibility of points which included facial and head points, facial and body' s points, facial、 head and body' s points). 3、 Determine MCV through electrophysiological method, stain the positive neurons of NGF and BDNF through immunochemical method and stain the positive neurons of BDNF mRNA through the method of ISH, all of
which made quantitative analysis through the expressive amounts of the positive cells. 4> Statistical analysis was done on SAS software. Analysis of orthogonal design was done by analysis of intuition and variance, compared among each group and in the group by analysis of variance, student-Newman-Keuls test and t-test.Results:In Rabbits had the symptom of leaning to right of nasal lip fosse, unfree movement with whisker and leaning back with body after model done in which MCV (MCV<10m/s) was slower than preceding model (P<0.01). 2> The comparation of MCV among all the group (except for control group) in preceding model was feasible which was not significant difference(P > 0.05) and the comparation of MCV among all the group (except for control group) after model done were feasible which was not significant difference(P> 0.05). 3> The restoration of MCV was faster and albumen of NGF and BDNF in facial nucleus and the level of expression of BDNF mRNA were higher in electroaccupuncture group than in model group after treatment in four weeks(P<0.01). 4> The restoration of MCV was fastest by selecting density wave shape combined facial and body' s points in acute period for treating experimental peripheral facial palsy. According the degree of curative effect influenced by each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture > compatibility of points. 5 -, The drop of expressive level of NGF albumen in facial nucleus was restrained at the most extent by selecting density wave shape combined faciah head and body' s points in acute period for treating experimental peripheral facial palsy. The expressive level influenced by each
factor from greatness to lightness was the wave shape of electroaccupuncture > the time of acupuncture > compatibility of points. 6^ The drop of expressive level of BDNF albumen in facial nucleus was restrained at the most extent by selecting density wave shape combined faciaK head and body' s points in acute period for treating experimental peripheral facial palsy. The expressive level influenced by each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture > compatibility of points. 7^ The drop of expressive level BDNF mRNA in facial nucleus was restrained at the most extent by selecting density wave shape combined faciaK head and body' s points in acute period for treating experimental peripheral facial palsy. The expressive level influenced by each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture> compatibility of points.Conclusions:1 n It had curative effect affirmatively on experimental peripheral facial palsy treated by electroaccupuncture with rabbits. It could lighten the swelling and denaturalization and reduce the death of neuron as well that could protect neuron and promote regeneration and restoration of function corresponding to facial nucleus after facial nerve injury in rabbits. 2> Different factor influence different curative effect of peripheral facial palsy treated by electroaccupuncture with rabbits. Each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture > compatibility of points. 3 > Through the method of orthogonal design, we concluded the best scheme for treating experimental peripheral facial palsy with rabbit was that acute period, selecting density wave shape and facial and body's
points. 4> NGF album n BDNF mRNA and album in facial nucleus as well as the drop of expressive level after facial nerve injury in rabbit was restrained and it would boost up the nutrition of opposite nerve which could protect neuron and take effect of the regeneration and restoration of facial nerve injury further through the method of density shape of electroaccupuncture combined faciaK head and body' s points in acute period. 5> Different stimulating parameters of acupuncture had different influence about the expression of neurotrophic factor. For instance, the choice of the wave shape of electroaccupuncture was the most important for the expression of NGF and the choice of the time of acupuncture was the most important for the expression of BDNF. 6> Orthogonal design was the highest effective design method of acupuncture's experiment about many factors and many levels approved by the study of experimental peripheral facial palsy treated by electroaccupuncture with rabbits. 7n Head' s points probably took effect of the regeneration and restoration after facial nerve injury through inhibit the drop of neurotrophic factor like NGF and BDNF in facial nucleus which could boost up the nutrition of facial nerve and promote the growth of facial neuraxon.
方法:1、选用日本大耳白兔66只,雌雄各半,采用神经卡压法造成左侧面神经损伤,作为研究实验性周围性面瘫的动物模型。2、66只兔随机分为11组(每组6只),其中一组作为假手术组,一组作为模型对照组,其余九组运用正交试验设计的方法,采用针刺时机(急性期、静止期、恢复期)、电针波形(密波、疏波、疏密波)、腧穴配伍(面部+头穴、面部+体穴、面部+头体穴)的三因素三水平搭配组合方案,随机分入L_9(3~4)正交表安排的九组实验中,进行实验观察,共观察四周。3、采用电生理的方法测定面运动神经传导速度(MCV);用免疫组织化学方法对面神经核神经生长因子(NGF)和脑源性神经营养因子(BDNF)阳性神经元进行染色,并用表达阳性细胞数定量分析;用原位杂交方法对面神经核脑源性神经营养因子(BDNF mRNA)阳性神经元进行染色,并用表达阳性细胞数定量分析。4、统计学分析在SAS for windows 8.0统计软件上完成。正交分析采用直观分析及方差分析,各组间、组内比较采用方差分析、q检验和t检验。
结果:1、造模后兔出现鼻唇沟偏向右侧,胡须不能自主运动,并向后倾斜等症状,且MCV小于10m/s,比造模前明显减慢(P<0.01)。2、各组(除假手术组)造模前MCV比较,无显著性差异(P>0.05),各组(除假手术组)造模后MCV比较,无显著性差异(P>0.05),各组间具有可比性。3、经四周治疗后,电针组的MCV恢复明显快于模型组(P<0.01)。电针组面神经核内NGF蛋白、BDNF蛋白、BDNF mRNA表达水平明显高于模型组(P<0.01)。4、采用急性期、选择电针疏密波形、取面部+体穴针刺治疗实验性周围性面瘫可以使MCV恢复最快,且各因素
Objective:To analyze the extent of curative effect influenced by all factors, optimize the therapeutic scheme of electroaccupuncture peripheral facial palsy and to discuss the influence of regeneration and restoration on facial nerve injury through different stimulating parameters of acupuncture which can provide the scientific experimental evidence for improving the clinical curative effect of peripheral facial palsy treated by acupuncture.Methods:1、 We made the animal model of experimental peripheral facial palsy with sixty-six rabbits of Japan species, male and female each half, by the method of crashing nerve. 2、 Sixty-six rabbits were divided eleven groups of which each group include six rabbits randomly. Of them one group was choused as control group, another as model group, others were divided nine groups experiment arranged by L9(3~4)orthogonal table for experimental observation for four weeks through combination scheme arranged in groups by three factors and three level , consisting of the time of acupuncture which included acute period, quiescence period and convalescence, the wave shape of electroaccupuncture which included dense wave, sparse wave and density wave, the compatibility of points which included facial and head points, facial and body' s points, facial、 head and body' s points). 3、 Determine MCV through electrophysiological method, stain the positive neurons of NGF and BDNF through immunochemical method and stain the positive neurons of BDNF mRNA through the method of ISH, all of
which made quantitative analysis through the expressive amounts of the positive cells. 4> Statistical analysis was done on SAS software. Analysis of orthogonal design was done by analysis of intuition and variance, compared among each group and in the group by analysis of variance, student-Newman-Keuls test and t-test.Results:In Rabbits had the symptom of leaning to right of nasal lip fosse, unfree movement with whisker and leaning back with body after model done in which MCV (MCV<10m/s) was slower than preceding model (P<0.01). 2> The comparation of MCV among all the group (except for control group) in preceding model was feasible which was not significant difference(P > 0.05) and the comparation of MCV among all the group (except for control group) after model done were feasible which was not significant difference(P> 0.05). 3> The restoration of MCV was faster and albumen of NGF and BDNF in facial nucleus and the level of expression of BDNF mRNA were higher in electroaccupuncture group than in model group after treatment in four weeks(P<0.01). 4> The restoration of MCV was fastest by selecting density wave shape combined facial and body' s points in acute period for treating experimental peripheral facial palsy. According the degree of curative effect influenced by each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture > compatibility of points. 5 -, The drop of expressive level of NGF albumen in facial nucleus was restrained at the most extent by selecting density wave shape combined faciah head and body' s points in acute period for treating experimental peripheral facial palsy. The expressive level influenced by each
factor from greatness to lightness was the wave shape of electroaccupuncture > the time of acupuncture > compatibility of points. 6^ The drop of expressive level of BDNF albumen in facial nucleus was restrained at the most extent by selecting density wave shape combined faciaK head and body' s points in acute period for treating experimental peripheral facial palsy. The expressive level influenced by each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture > compatibility of points. 7^ The drop of expressive level BDNF mRNA in facial nucleus was restrained at the most extent by selecting density wave shape combined faciaK head and body' s points in acute period for treating experimental peripheral facial palsy. The expressive level influenced by each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture> compatibility of points.Conclusions:1 n It had curative effect affirmatively on experimental peripheral facial palsy treated by electroaccupuncture with rabbits. It could lighten the swelling and denaturalization and reduce the death of neuron as well that could protect neuron and promote regeneration and restoration of function corresponding to facial nucleus after facial nerve injury in rabbits. 2> Different factor influence different curative effect of peripheral facial palsy treated by electroaccupuncture with rabbits. Each factor from greatness to lightness was the time of acupuncture > the wave shape of electroaccupuncture > compatibility of points. 3 > Through the method of orthogonal design, we concluded the best scheme for treating experimental peripheral facial palsy with rabbit was that acute period, selecting density wave shape and facial and body's
points. 4> NGF album n BDNF mRNA and album in facial nucleus as well as the drop of expressive level after facial nerve injury in rabbit was restrained and it would boost up the nutrition of opposite nerve which could protect neuron and take effect of the regeneration and restoration of facial nerve injury further through the method of density shape of electroaccupuncture combined faciaK head and body' s points in acute period. 5> Different stimulating parameters of acupuncture had different influence about the expression of neurotrophic factor. For instance, the choice of the wave shape of electroaccupuncture was the most important for the expression of NGF and the choice of the time of acupuncture was the most important for the expression of BDNF. 6> Orthogonal design was the highest effective design method of acupuncture's experiment about many factors and many levels approved by the study of experimental peripheral facial palsy treated by electroaccupuncture with rabbits. 7n Head' s points probably took effect of the regeneration and restoration after facial nerve injury through inhibit the drop of neurotrophic factor like NGF and BDNF in facial nucleus which could boost up the nutrition of facial nerve and promote the growth of facial neuraxon.
引文
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