| Objective: Three-week-old cerebral palsy rats were used as subjects.CatWalk XT gait analysis,VBM,ALFF and immunohistochemistry were used to study the brain structure,function and the effects of M1,MCT1,CytC,Caspase-3 in treatment with bagua scalp acupuncture,which compared with the scalp acupuncture named international standardization program.In this way we attempt to verify the bagua scalp acupuncture in treating movement disorder of cerebral palsy is of more effective,then analysis the mechanism of brain structures,functions and the related proteins in order to provide scientific basis to establish this new scalp acupuncture.Method and Results: Part one: Effect of bagua scalp acupuncture on the motor function of CP rats.Method: 60 healthy male Wistar rats aged 3 weeks were randomly divided into normal group,acupuncture group 1,acupuncture group 2,sham acupuncture group,model group and sham operation group.We used the classic HIE model to make CP rats,that is,the left common carotid artery was ligated to cause permanent interruption,then placed the rats in a hypoxic environment with an oxygen concentration of 8% for 2 h.The normal group is healthy and no treatment on it,the acupuncture group 1,acupuncture group 2,sham acupuncture group and model group all accepted HIE operation,and there is only a neck skin cutting on sham operation group which without common carotid artery ligated and hypoxia.Acupuncture 1 group used bagua scalp acupuncture,acupuncture 2 group used scalp acupuncture named international standardization,sham acupuncture group used non-acupoint acupuncture to treat,which need leaving needle for 20 minutes,performing 6 times a week and lasting for 2 weeks.The normal group,the model group and the sham operation group were normally grasped daily without any other intervention.The gait analysis was performed before modeling,after modeling,seven days of treatment and 14 days of treatment,and the suspension test,Grip test and posture reflex test were performed before modeling,after modeling,and 14 days of treatment.Results: 1.The suspension test and the Grip test at 14 days showed that the model group,the acupuncture group 2,the sham acupuncture group had lower score than the normal group and the sham operation group,and the difference was statistically significant(P<0.05).There was no significant difference between the acupuncture group 1 and the acupuncture group 2(P>0.05),but the acupuncture group 1 suspension test and the Grip test score were higher than the acupuncture group 2,suggesting that the acupuncture group 1 has better tendency than the acupuncture 2 group in improving of the suspension test and Grip test score of CP rats.2.The results of posture reflex test at the 14 days showed that the model group,the acupuncture group 1,the acupuncture group 2 and the sham acupuncture had lower scores than the normal group,and the difference was statistically significant(P<0.001).The acupuncture group 1 compared with the acupuncture group 2,the scores were increased,and the difference was statistically significant(P<0.001),suggesting that the acupuncture group 1 was better than the acupuncture group 2 in improving the posture reflex abnormality of CP rats.3.The average speed of CatWalk gait analysis at 14 days showed no significant difference between the groups(P>0.05),suggesting that the movement speed of CP rats improved with time.The average speed of acupuncture group 1 has higher tendency than that model group,acupuncture group 2,suggesting that the acupuncture group 1 had better tendency to improve the movement speed of CP rats than the acupuncture group 2.4.The average pressure of the left forelimb after 7-day treatment showed that the average pressure of the left forelimb in the acupuncture group 1 was lower than that in the acupuncture group 2,the difference was statistically significant(P<0.05),suggesting that in the acute period acupuncture group 1 has a better intendency of improvement in restoring compensatory raising of the average pressure of left posterior forelimb than that of the acupuncture 2 group.The average pressure analysis of the left forelimb after 14-day treatment showed that the average pressure of the left forelimb in the model group,the acupuncture group 2 and the sham acupuncture group was higher than that in the normal group(P<0.05).The average pressure of the left forelimb in acupuncture group 1 was lower than that of the model group(P<0.05).There was no significant difference between the acupuncture group 1 and the acupuncture group 2(P>0.05),but the average pressure of the left forelimb in the acupuncture group 1 has lower tendency than the acupuncture group 2,which suggests that the acupuncture group 1 has better tendency to compensate the enhancement in the average pressure of the left forelimbs of CP rats than the acupuncture group 2.5.The average pressure of right forelimb in treatment after 14 days showed that the model group and the sham acupuncture group was lower than that in the normal group(P<0.05).The average pressure of the right forelimb in the acupuncture group 1 was higher than acupuncture 2 group,suggesting that the acupuncture group 1 has a better tendency to recover the average pressure of the right forelimb of CP rats than the acupuncture group 2.Part two: The effect of the brain structure and function of CP rats with bagua scalp acupuncture Method: 60 healthy Wistar rats aged 3 weeks were randomly divided into normal group,acupuncture group 1 and model group.Modeling and intervention methods are the same as in the first part.The functional magnetic resonance imaging data were collected after 14-day treatment,then analysis the gray matter volume and ALFF of the three groups.Results: 1.After 14 days of modeling,the gray matter volume of the model group and the normal group showed that in the left cerebellar lobule,the olfactory cortex,the parasitoid,the hippocampus,the first somatosensory cortex,the amygdala,the gray matter volume of the outer capsule,the first somatic motor cortex,the frontal cortex,the corpus callosum,the cingulate cortex and the right temporal lobe cortex,the model group was significantly smaller than that of the normal group,the difference was statistically significant(P<0.001).In the bilateral cerebellar lobules,the right dorsal cortex,insular cortex and temporal cortex,the gray matter volume was significantly bigger than that in the normal group,and the difference was statistically significant(P<0.001).2.After 14 days of treatment,the gray matter volume of the acupuncture group 1 and the normal group showed that in the left flank,the single leaflet,the granule cell cortex,the external olfactory cortex,the hippocampus,and the dorsolateral nucleus of the thalamus,globus pallidus,internal sac,right upper parietal gray matter layer,hypothalamic nucleus,striatum(caudal nucleus),first somatosensory cortex(upper labial region),bilateral external capsule,cingulate cortex 1 area,etc,the gray matter volume of the acupuncture group 1 was significantly smaller than that of the normal group,and the difference was statistically significant(P<0.001).The gray matter volume in the right trigeminal ridge and the left cerebellar lobules was significantly larger than that in the normal group 1(P<0.001).3.After 14 days of treatment,the gray matter volume of the acupuncture group 1 and the model group showed that in the right olfactory cortex,temporal lobe cortex and second optic cortex(outer area),the gray matter volume of the acupuncture group 1 was significantly larger than that of the model group.The difference was statistically significant(P<0.001).The gray matter volume of the model group in the right vertebral region was significantly larger than those in the acupuncture group 1,and the difference was statistically significant(P<0.001).4.14 days after modeling,the results of ALFF analysis showed that in the normal group the ALFF values in the bilateral dorsolateral cortex and the left insular cortex were significantly lower than that in the normal group,and the difference was statistically significant(P<0.05).In bilateral thalamus,left hy Pothalamus,left almond(pear-shaped area),left dentate gyrus,left first somatosensory cortex(barrel/upper lip area),the hippocampus,right midbrain deep nucleus and right first visual cortex,the acupuncture group 1 were significantly higher than those in the normal group(P<0.01).5.After 14 days of treatment,the results of ALFF in the acupuncture group 1 and the normal group showed that the ALFF value of the acupuncture group 1 was significantly lower than that of the normal group in the bilateral cingulate cortex,the right hippocampal olfactory island,and the left lateral ventricle.The difference was statistically significant(P<0.05).In bilateral hippocampus,bilateral amygdala(intracortical,base),bilateral first somatosensory cortex(barrel,upper lip),bilateral frontal,right paracentral lobule,right reticular nucleus,right facial nerve root,right central gray matter,right first visual cortex,right pons nucleus,right first auditory cortex,right dentate gyrus,right external capsule,the right globus pallidus,the right island leaf,the left trigeminal nucleus,the left vertebral body,the left hypothalamic nucleus,the left midbrain deep nucleus,the left internal sac,and the left motor cortex,acupuncture group 1 has a higher ALFF values than the normal group,the difference was statistically significant(P < 0.05).6.After 14 days of treatment,the results of ALFF in the acupuncture group 1 and the model group showed that the ALFF value of the model group in the left vertebral body area was significantly lower than that of the acupuncture group,the difference was statistically significant(P<0.05).The ALFF value of a motor cortex,the right posterior thalamic nucleus and the right first somatosensory cortex(barrel/small area/forelim area/hind limb area)was significantly higher than that of the acupuncture group 1,and the difference was statistically significant(P <0.05).Part three: Effect of the expression of MCT1,CytC and Caspase-3 in CP rats with bagua scalp acupuncture Method: 30 male Wistar rats aged 3 weeks were randomly divided into normal group,model group,acupuncture group 1,acupuncture group 2 and sham acupuncture group.Modeling and intervention methods are the same as in the first part.After 14 days of treatment,the brain was taken immunohistochemically detected.The expressions of MCT1,CytC and Caspase-3 in the M1 region of the brain of each group were calculated and analyzed.Results: 1.The expression of MCT1 protein in the brain tissue of the model group,acupuncture group 1,acupuncture group 2 and sham acupuncture group was higher than that in the normal group(P<0.05).Among them,the expression of MCT1 protein in acupuncture group 1,acupuncture group 2 were higher than that in model group(P<0.05).The expression of MCT1 protein in acupuncture group 1 was higher than that in acupuncture group 2(P<0.05).2.The expression of CytC protein in the M1 area of the model group and the acupuncture group 1 were higher than that of the normal group(P<0.001).The acupuncture group 2 and the sham acupuncture group were higher than that of the normal group.(P<0.01).Compared with the model group,the expression intensity of CytC protein in acupuncture group 1 and acupuncture group was lower than that in the model group.The difference was statistically significant(P<0.01).There was no significant difference in the expression of CytC protein between acupuncture group 1 and acupuncture group 2(P>0.05),but the expression of MCT1 protein of acupuncture group 1 has decreased tendency compared with acupuncture group 2.3.The expression of Caspase-3 protein in the brain tissue of the model group,acupuncture group 1,acupuncture group 2 and sham acupuncture group was higher than that of the normal group(P<0.001).The expression of Caspase-3 protein in acupuncture group 1 was lower than that in the model group(P<0.001).The expression intensity of Caspase-3 protein in acupuncture group 1 was lower than that in acupuncture group 2,the difference was statistically significant(P<0.001)Conclusion: 1.The bagua scalp acupuncture method is superior tendency in improving overall movement,bearing capacity and coordination than that of to the scalp acupuncture named international standardization.2.The bagua scalp acupuncture method has the effect of promoting the neuronal activity in the M1 region and the lesion-side vertebral region of CP rats,and also has neuronal activity in the infoliate island,vertebral body and bilateral hippocampus.Certainly,it can improve the motor dysfunction of CP rats.3.The bagua scalp acupuncture method promotes the expression of MCT1 protein and inhibits the expression of CytC and Caspase-3 protein in the M1 region of the lesion side,thereby promoting energy metabolism,inhibiting apoptosis and repairing M1 neurons in the M1 region of the lesion side. |
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