| Objective:Based on the previous research,this study takes the rats with facial nerve crush injury as the research carrier.Starting from the important parameters of electro-acupuncture quantity of stimulus,i.e.intensity,frequency,time and course of treatment,setting three quantity of stimulus—mild,moderate and severe,then screening the effective stimulus for using electro-acupuncture to treat peripheral facial paralysis rats.Designed to provide a scientific experimental basis for modern research on the classical theory of traditional Chinese medicine,which is the disease lasts’.Method:Experiment 1:Facial paralysis model was made by crushing the main trunk of facial nerve.Forty Sprague-Dawley(SD)male rats were randomly divided into five groups according to the difference of mold crushing time and methods.There were 8rats in each group.The groups include blank group,1 min A group(press 30 s for front side,release 30 s,press 30 s for back side),1 min B group(Simply press 1min),3 min group(Simply press 3min)and 5 min group(Simply press 5min).The self-healing of rats in each group was observed within 14 days without any intervention.Then Using Simone 10 method to do praxiology grade in 1,7,14 day.After the model was established 14 days later,using the HE stain to abserve the histomorphological changes of injured facial nerve,compare the differences of injury among groups,and record praxiology score,facial nerve grade.Finally screen a method which has a long self-healing cycle and heavy injury in establishing facial nerve crush injury model in rats.We can use this model to do preparation for the objective evaluation of the therapeutic effect of electroacupuncture in the next step.Experimentation 2:This experiment is based on the 3*5 factorial design of two factors,namely the amount of stimulation and the course of treatment.90Sprague-Dawley(SD)male rats were randomly divided into five groups according tothe difference parameters of stimulus,namely blank group,model group,mild stimulation group,moderate stimulation group and severe stimulation group.Each group was further divided into three subgroups by course of treatment(7 days,14 days,21 days),totally 15 groups,6 rats in each group.The model crushing method selected from experiment 1.Acupoints were acupunctured: Hegu,Dicang,Jiache,Yifeng.The operating site is the affected side.The waveform(density wave),intensity(1Hz),total treatment time(30 minutes)were fixed.Then every groups be treated according to the corresponding parameters of stimulus.7 days as a course of treatment,treating once daily,continuous treatment for 5 days,then rest for 2 days.The model and black groups were fixed and bound by the same method.The injured facial nerve and facial muscle were harvested at the end of the corresponding course of treatment.The morphological changes of facial nerve and facial muscle were observed by HE staining under light microscope.Recording the praxiology score,facial nerve grade,and the diameter of facial muscle fiber to screen the effective quantity of stimulus for the treatment of this disease by electro-acupuncture.Results:Experiment 1:According to the results of behavioral score,1 day after models established,the scores of the four different pressing time groups were all lower than 4 points,and four groups scores were all significantly lower than blank group(P<0.01).After 7 days and 14 days of models established,the score of group 1minB decreased(P<0.05),group 1minA,group 3min and group 5min all significantly decreased(P < 0.01)compared with the blank group.there was no statistical significance between the four different groups.14 days after the models established,the results of facial nerve grade showe:compared with the blank group 1min B group grade was lower(P>0.05),both 3min and 5min two group facial nerve grade were all higher than blank group(P< 0.05),1 min A group grade was highest(P > 0.05).There were no statistical significance between 1 min A group,3 min group and 5 min group(P>0.05).Experiment 2:1.According to the results of behavioral score,the scores of the three treatment courses of the model group were all reduced compared with the blank group(P<0.05).Compared with the model group,electroacupuncture scores of each group 1,2and 3 courses were all higher than that of the model group(P<0.05).The mild group was restored to the blank group after 2 courses(P > 0.05),and three groups were restored after 3 courses(P>0.05).Three electroacupuncture groups appeared in all three sessions:There was no significant difference between the three groups(P>0.05).There was no statistically significant difference between 7d group and 14 d group,and between 14 d group and 21 d group,while there was statistically significant difference between 7d group and 21 d group(P<0.05).2.According to the results of facial nerve grade,three treatment courses of the model group and severe group were all higher than blank group(P<0.05),The mild group recovered to no difference with the blank group after 1 course of treatment,and medium group recovered after 3 courses.three courses of mild group had lower grading than model group(P<0.05),and there was no difference in 1 and 2 courses of the medium group compared with model group,but 3 courses of the medium group had lower grading compared with model group(P<0.05).there was no difference in the three courses of severe group compared with model group.Comparison among the three groups of electroacupuncture: there was no difference among the three groups after 1 course,and the grading of severe group was higher than mild group after 2courses(P<0.05),the grading of severe group was higher than mild and moderate groups after 3 courses(P<0.05).There was no difference between the three treatment courses of severe group,and the scores of the moderate and mild groups were increased between 7d group and 14 d group,and between 7d group and 21 d group(P<0.05),while 21 d group were not significantly different from 14 d group(P>0.05).3.The results of the diameter of facial muscle fibers showed that:compared to the blank group,the diameter of facial muscle in model group was decreased in 1 and2 courses of treatment(P<0.05),then 3 courses recovered.Three electroacupuncture groups compared with the model group:there was no significant difference betweeneach group and model group after 1 course.After 2 courses of mild group,the diameter increased(P<0.05)and recovered to the same level as that of the blank group.There was no significant difference in the diameter between medium and sever groups compered with the model group(P>0.05).After 3 courses of treatment,there was no significant difference between the mild and moderate groups compered with model group(P>0.05),and the sever group increased significantly(P<0.05).There was no difference among the three treatment courses of medium stimulation.Blank group,model,mild and sever 3 course of each groups all appeared :with the increase of time,the diameter of the facial muscle increased,diameter of 21 d group was higher than 7d group(P<0.05).Conclusion:1.The facial paralysis model of rats can be successfully established through nerve crushing models with different pressing time.The method of 1 min A group(30seconds for crushing frontal,30 seconds for loosening and then crushing again in 30seconds)is the best.2.Electroacupuncture stimulation can accelerate the recovery process of facial nerve injury and promote the morphology of facial nerve to return to normal.However,excessive stimulation can cause injury,so we should follow the principle of "the disease lasts".3.The effective stimulation amount of electroacupuncture treatment for this disease is mild stimulation(density wave 1Hz,1mA stimulation intensity),14 days(2courses of treatment),electroacupuncture stimulation for 10 minutes,needle retention for 20 minutes. |
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