| Background: Peripheral nerve injury is a common clinical disease,which often leads to physical dysfunction such as exercise and sensation of the patient,which seriously affects the daily activities of the patient and hinders the participation of the activity,which leads to a decrease in the quality of life and an increase in social burden.How to promote its functional recovery has always been a key issue to be solved urgently in clinical practice.Vibration therapy has been proven to improve limb muscle strength and promote neurological recovery.However,the vibration parameters used in the treatment of disease by vibration therapy are not the same,and there is no basis for selection.Based on previous research,this research group found that frequency is one of the key factors in the effect of vibration treatment.Each organism is an elastic body.When it vibrates,it can be regarded as a spring-mass-damping system.Each organ has its inherent The frequency exists.Therefore,this experiment will set the vibration parameters less than,equal to,and greater than the natural frequency based on the measured natural frequency,and apply it to the rat model of sciatic nerve clamp for local vibration therapy,using behavioral,electrophysiological,Histomorphology and other methods were used to observe the recovery of neurological function in rats,and the potential effect pathways of limb function recovery after peripheral nerve injury were explored by immunohistochemistry,which provided a theoretical basis for vibration therapy to promote the recovery of peripheral nerve injury.Part I: Determination of the natural frequency of gluteus maximus and calf triceps in rats after sciatic nerve injuryObjective: To establish a three-degree injury model of rat sciatic nerve,and to determine the natural frequencies of gluteus maximus and calf triceps in rats after injury,and to observe whether the natural frequencies of the same parts in different states are consistent,and provide an experiment for the selection of vibration therapy frequency.in accordance with.Methods: A total of 36 Sprague-Dawley male rats(clean grade)were randomly divided into normal group(n=14),model group(n=14)and anesthesia group(n=8)for natural frequency measurement.The model group underwent a sciatic nerve clamp injury model.The anesthesia group was anesthetized with abdominal cavity,and no treatment was done.?1 Model identification: After modeling,6 rats in the normal group and the model group were randomly selected for Sunderland III degree nerve injury model identification,and determined by motor nerve conduction velocity and sciatic nerve HE staining;?2 natural frequency detection: 7th after modeling On the basis of vibration excitation instrument,the rat gluteus maximus and calf triceps muscle were subjected to shock vibration excitation.Based on the data measured by the vibration response detection module,the rat gluteus maximus was determined according to real-time spectrum observation and time-frequency analysis method.The natural frequency of the triceps of the calf.Results:(1)Model identification: Electrophysiological detection: The model group met the MNCV drop to below 10m/s,and the action potential amplitude decreased.HE staining: normal group: the nerve fibers are arranged neatly,consisting of nerve bundles of different sizes.The nerve bundles are surrounded by the nerve bundle membrane and the outer membrane.The nerve tissue structure is complete and evenly distributed.The model group: the nerve fibers are broken and arranged disorderly.The interstitial blood vessels are congested,the tissue structure is loosely arranged,and the nerve bundle membrane remains continuous.(2)There were two regular peaks in each of the three groups of rats.The comparison results were as follows: 1)The first peak frequency of gluteus maximus and calf triceps in the blank group,model group and anesthesia group Statistical difference(P>0.05);2)There was no significant difference in the second peak frequency of gluteus maximus and calf triceps in the blank group,model group and anesthesia group(P>0.05).3)There was no significant difference in frequency between gluteus maximus and calf triceps(P>0.05).Conclusion:(1)The sciatic nerve III degree injury model was successfully established.(2)Establishing the first peak of rat gluteus maximus and calf triceps as its natural frequency;the natural frequency of gluteus maximus and calf triceps muscle tissue is stable,and the natural frequency of the same muscle tissue is not due to its The state is different and there is a big difference.Part II: Behavioral and electrophysiological studies on rat sciatic nerve injury after different frequency vibration therapyObjective: To evaluate the effects of different frequency of local vibration therapy on nerve connectivity and functional recovery of hind limbs in rats after sciatic nerve injury by animal behavioral and nerve conduction velocity observations.Methods: A total of 48 Sprague-Dawley male rats(clean grade)were randomly divided into sham operation group(n=8),model group(n=8),vibration treatment group(n=24),and sham intervention group(n=8).The sham operation group only exposed the sciatic nerve but did not clamp;the model group,the vibration treatment group,and the sham intervention group all established the right sciatic nerve clamp injury model.There was no special intervention in the sham operation group and the model group;the vibration treatment group was vibrated at the central position of the gluteus maximus muscle at 3Hz,8Hz,and 13 Hz respectively;the dummy intervention group only placed the head end of the vibrator on the gluteus maximus muscle of the affected side,the center of the abdomen is not vibrating.Because the Catwalk gait analysis of the injured limbs was not detected at 1 week after surgery,the sham operation group could heal 1 week after surgery,and the experimental treatment was performed on the 7th day after nerve injury,so Cat Walk gait analysis was performed on 1 week,2 weeks,3 weeks,and 4 weeks after treatment respectively,in order to observe changes in maximum contact average intensity,footprint area,and swing speed of rats.The threshold of mechanical contraction response was 7 days before and after surgery.The rats were tested at 1 week,2 weeks,3 weeks,and 4 weeks to evaluate the effects of local vibration therapy on behavioral motor and sensory function after sciatic nerve injury in rats,and the neuroelectrophysiology(sciatic nerve conduction)was measured at 4 weeks.Speed),to evaluate the effect of local vibration therapy on neural connectivity after rat isthmus injury.Results:(1)Gait analysis-The maximum contact mean intensity results showed that the 3Hz group was higher than the 13 Hz group,the sham intervention group and the model group(P<0.05)at the first week of treatment,and reached the maximum at the second week.There were differences between the 8Hz and 13 Hz groups,the false intervention group and the model group(P<0.05),and remained stable in the third week,which was different from the 8Hz,13 Hz group,the false intervention and the model group(P<0.05),and declined slightly at the forth week.The 8Hz group was higher than the 13 Hz group,the sham intervention group and the model group at the first week of treatment(P<0.05),and remained stable at 2,3,and 4 weeks.The increase in the first and second weeks of the 13 Hz group was small,and it rose steadily in the third and fourth weeks.There was no significant difference between the model group and the sham intervention group(P>0.05).By the fourth week,the 3Hz group,the 8Hz group and the 13 Hz group were significantly higher than the sham intervention group and the model group(P<0.05),but they did not reach the level of the sham operation group(P<0.001).(2)Gait analysis-The print area results showed that the 3Hz treatment group showed a significant upward trend at the first and second weeks,reaching the highest level at the second week,and was statistically different from the 13 Hz group,the sham intervention group,and the model group(P<0.05),the third week was still higher than the sham intervention group and the model group(P<0.05),and the third and fourth weeks showed a steady trend.The 8Hz treatment group showed an upward trend at the first and second weeks,and was significantly higher than the sham intervention group and the model group at the second week(P<0.05).The second and third weeks showed a slight downward trend,and the third and fourth weeks increased again.The uptrend trend of the 13 Hz group was lower than that of the 3Hz group and the 8Hz group at the first and second weeks,but it increased significantly at the third and fourth weeks,and was statistically different from the model group and the sham intervention group(P<0.05).By the fourth week,the 3Hz group,the 8Hz group and the 13 Hz group were higher than the model group and the sham intervention group(P<0.05),but they did not reach the level of the sham operation group(P<0.001).(3)Gait analysis-The swing speed results showed that the 3Hz treatment group showed an upward trend at the first and second weeks,which were higher than the model group and the sham intervention group(P<0.05),and at the 2nd week with the 13 Hz,sham intervention group,there was a statistical difference in the model group(P<0.05),which continued to increase in the second and third weeks,and remained stable in the third and fourth weeks.The first and second weeks of the 8Hz treatment group were higher than the model group and the sham intervention group(P<0.05),and continued to increase in the second and third weeks,but the slope was lower than that in the 3 Hz group,and remained stable in the third and fourth weeks.The increase of the 13 Hz group was lower than that of the 3Hz and 8Hz groups at the first and second weeks,but higher than that of the model group and the fake intervention group(P<0.05),the second,third and fourth weeks showed a significant upward trend,and reaching the highest level at the fourth week.Although the 3Hz,8Hz,and 13 Hz vibration treatments were significantly improved,they did not reach the level of the sham operation group(P<0.001).(4)The paw withdraw threshold test statistics showed that the baseline of the rats in each group was the same before operation,and the pain threshold of the rats was significantly decreased after operation.There was significant difference between the groups and the sham operation group(P<0.001).3Hz,8Hz,13 Hz have an upward trend,8Hz,13 Hz rising trend is obvious and statistically different from the model group and the fake intervention group(P<0.05);the first and second weeks of treatment,3Hz,13 Hz continue to rise,8Hz is stable The trends were statistically different from the model group and the sham intervention group(P<0.05).At the 2nd and 3rd week of treatment,3Hz,8Hz and 13 Hz showed an upward trend,which was statistically different from the model group and the sham intervention group(P<0.05),13 Hz reached the highest amplitude at the 3rd week;13Hz and 8Hz remained stable and decreased slightly at the 3rd and 4th week of treatment,while the 3Hz group continued to rise,reaching the maximum at the 4th week,during the period with the model group,There was a statistical difference in the sham intervention group(P<0.05).The 3Hz group,the 8Hz group,and the 13 Hz group were significantly improved at 4 weeks of treatment but did not reach the level of the sham operation group(P<0.05).(5)Electrophysiological statistics showed that after 4 weeks of intervention,the motor nerve conduction velocity(MNCV)of the model group,the sham intervention group,the 3Hz group,the 8Hz group,and the 13 Hz group was significantly lower than that of the sham operation group(P<0.001);The MNCV of the 3Hz group,the 8Hz group,and the 13 Hz group was significantly higher than that of the model group and the sham intervention group(P<0.05).There was no significant difference between the 3Hz,8Hz and 13 Hz group(P>0.05),but the 3Hz group was slightly higher than the 8Hz and 13 Hz groups.Conclusion: Different frequency local vibration therapy can improve the gait analysis parameters maximum contact mean intensity,print area and swing speed of sciatic nerve clamp in rats,relieve pain sensitivity and improve motor nerve conduction velocity after sciatic nerve clamp injury,and 3Hz is better than 8Hz and 13 Hz.Part III:Immunohistochemical study of histomorphology and neuro-related factors in rats with sciatic nerve injury by different frequency vibration therapyObjective: To investigate the effects of different frequency local vibration therapy on the morphology of regenerative nerve tissue and the expression of nerve growth related factors in rats after sciatic nerve injury by electron microscopy and immunohistochemistry.Methods: A total of 144 male Sprague Dawley rats(clean grade)were randomly divided into model group(n=24),treatment group(n=72),sham intervention group(n=24),sham operation group(n=24),and the first four groups established a right sciatic nerve clamp injury model.The treatment group was given 3Hz,8Hz,13 Hz frequency vibration on rat gluteus maximus muscle center,2 min/time,1 time/day,no special intervention in the model group and the normal group.At the 1st,2nd,and 4th weeks after treatment,the distal nerves of the injured site were observed by electron microscopy and semiquantitative analysis of immunohistochemistry.The morphology of regenerative nerve tissue and the nerve growth-related factor protein expression were evaluated after the sciatic nerve injury in rats.Results:(1)Electron microscopic observation: Waller degeneration occurred in each group 1 week after treatment.The myelin fibers were obvious in the model group and the sham intervention group,the lamellar structure was blurred,the gap was widened,and the new myelin sheath was seen in each group.The myelin sheath was thinner.The new myelin sheath in the treatment group was more than the model group and the morphology was regular.It suggested that the nerve entered the regeneration stage at 1 week of treatment,and the axial filament increased and grew distally.At 2 weeks,the model group and the pseudo-intervention group the sheath was not compact,the shape was irregular,and the myelinated nerve fibers were flat.The myelin sheath increased in the treatment group,and the myelin density was higher than that in the model group and the sham intervention group.At 4 weeks,the thickness of myelin sheath was thinner in model group and sham intervention group,the myelination of regenerated nerve fiber was incomplete,demyelination was present and the density of myelin sheath was low.The distribution of myelinated nerve fibers in each treatment group was more than that in model group and sham intervention.The density of myelin sheath was higher than that of the model group and the sham intervention group.The myelin structure of the regenerated nerve fibers was more uniform.Schwann cells were observed.The morphology of the tissues in the treatment group did not change much,but the 8 Hz demyelination phenomenon is more than 3 Hz and 13 Hz at 4 weeks.The results suggest that vibration therapy can promote the recovery of regenerative nerve morphology after sciatic nerve injury in rats.(2)S-100 nerve expression: 1 week after treatment,the average optical density(AOD)of S-100 protein in the vibration treatment group was higher than that in the sham intervention group(P<0.05),but there was no treatment between 3 Hz,8 Hz and 13 Hz in the treatment group.The difference was statistically significant(P>0.05).There was a statistical difference between the treatment group and the sham intervention group(P<0.001).The expression of S-100 was higher in 3Hz than in 8Hz and 13 Hz in the treatment group.There was a statistical difference between the two groups(P<0.05).After treatment for 4 weeks,the S-100 protein content in the treatment group remained high,which was statistically different from the sham intervention group(P<0.001).In the treatment group,the expression of 3Hz S-100 was higher than 8Hz and 13 Hz,which showed a trend of continuous increase,but there was no statistical difference compared with 8Hz and 13Hz(P>0.05).In the same group,the S-100 expression in the sham intervention group was statistically different from the first week and the second week and the fourth week(P<0.05),but there was no significant difference between the second week and the fourth week(P>0.05).The expression of S-100 in each treatment group also proliferated actively with the prolongation of time.There was significant difference between the first week and the second and fourth weeks(P<0.05),and there was no significant difference between the second and fourth weeks(P>0.05).(3)BDNF nerve expression: After treatment for 1 week,the expression of BDNF in the treatment group was higher than that in the sham operation group(P<0.05).There was no significant difference between the treatment group at 3Hz,8Hz and 13Hz(P>0.05);There was significant difference between the treatment groups and the sham intervention group(P<0.05).The treatment group had the highest rise at 13 Hz in the second week,but there was no statistical difference between 3Hz,8Hz and 13Hz(P>0.05).After 4 weeks of treatment,each treatment group maintained a high level,compared with the sham intervention group(P<0.05),and the 3Hz group showed strong positive at the 4th week,and the difference was statistically significant compared with the 8Hz group(P<0.05).In the same group,there was no significant change in the time of the sham intervention group(P>0.05).There was no significant difference in the expression of BDNF at 3Hz between the first and second weeks(P>0.05),but there was a statistical difference between the two groups and the fourth week(P<0.05);8Hz BDNF expression was statistically different at 1 week,2 weeks,and 4 weeks(P<0.05),and there was no significant difference between the 2nd week and the 4th week(P>0.05);the BDNF expression in 13 Hz group and the trend of the 8Hz group was basically the same,and there was a statistical difference between the first week and the 2nd and 4th week(P<0.05).There was no significant difference between the 2nd week and the 4th week(P>0.05).(4)CNTF nerve expression: There was no significant difference in CNTF expression between the treatment group and the sham-treated group(P>0.05).After 2 weeks of treatment,the expression of CNTF in each treatment group was significantly higher than that in the sham-treated group and the difference was statistically significant(P<0.05),but there was no significant difference between the treatment group at 3Hz,8Hz and 13Hz(P>0.05).After 4 weeks of treatment,each treatment group maintained a steady upward trend,and there was statistical difference compared with the sham intervention group(P<0.05).At 4 weeks,the CNTF expression of the 3Hz group was higher than that of 8Hz and 13 Hz.In the same group,there was no significant difference in the sham intervention group at the 1st and 2nd week(P>0.05),but there was a statistical difference between the 1st and 2nd week and the 4th(P<0.05),suggesting that CNTF expression may be slowly recover as time goes by.The expression of CNTF in the 2nd and 4th weeks of the 3Hz group was higher than 1 week(P<0.05),but there was no significant difference between the 2nd week and the 4th week(P>0.05).The CNTF expression in the 8Hz and 13 Hz groups was higher at the 2nd and the 4th weeks than the first week,and there was statistical difference(P<0.05),but there was no statistical difference between the two time points at 2 weeks and 4 weeks(P>0.05),and the highest expression at 3Hz was observed at the 4th week and present upward trend.Conclusion: Different frequency vibration therapy can promote the morphological recovery of regenerative nerve tissue in rats with sciatic nerve clamp injury;increase the expression of S-100 protein in Schwann cells,promote the secretion of neurotrophic factors BDNF and CNTF,and 3Hz is better than 8Hz and 13 Hz. |
PDF Full Text Request