| ObjectiveObserve the effect of the cluster needling of scalp acupuncture on the neurological function score and ultrastructure of ischemic penumbra at different time points in rats with acute focal cerebral ischemia.From the perspective of the ultrastructural,explore the possible mechanism of scalp acupuncture treatment for cerebral infarction.Methods54 healthy clean-grade male SD rats were randomly divided into three groups,named the false operation group(A,n=18),model group(B,n=18),and the cluster needling of scalp acupuncture group(C,n=18).Every group was divided into three groups(6 rats in each group)according to time points of 24h,7d,14d.The local cerebral ischemia rats model in Group B and C were prepared by modified suture-occluded method,while Group A was sham model.Rats in Group C were treated by the cluster needling of scalp acupuncture,by contrast,Group A and B were intervened without acupuncture treatment.Once a day,treatment 6d,rest 1d,7d for a course of treatment.The rats in each group were evaluated by Bederson's scoring system after being operated and being killed.The ultrastructure of the nerve of each group was tested by transmission electron microscope.Results1 neurological score:Intra-group comparison:The neurological score in group C and B 4h after the rat waking compared with that in group A was statistically significant different(P<0.05),which showed that the MCAO model was successfully conducted and models have comparability.However there was no significant between group C and group B(P>0.05).On the time point of 24h and 7d after the rat waking,there was no significant difference between group C and group B(P>0.05).However,on the time point ofl4d,the neurological score in group C was lower than that in group B,we can found a statistically significant difference(P<0.05).Inter-group comparison:In group C,the neurological score at the time point of 14d was lower than that at the time point of 24h,also there was a statistically significant difference(P<0.05).We can found that there was no statistically difference(P>0.05)of the neurological score between the time point of 14d and the group of 7d.There was no statistically difference(P>0.05)of the neurological score between the time point of 7d and 24h.In the group B,the neurological score at the time point of 14d was lower than that at the 24h and 7d,the three groups also didn't show any statistically difference(P>0.05).2 Ultrastructural morphological changesIn group A,there was no significant difference in morphological structure between the time point of 24h,7d and 14d.There are abundant of neuronal organelles,and the nucleus morphology is regular.The glial cells have normal morphology and structural integrity.There are abundant of synapses and structural integrity.The presynaptic membrane is rich in synaptic vesicles and ribosomes.On the time point of 24h,in group B,endothelial thickening,poor vascular endothelial cells,inflammatory cells exuding in the blood vessels,severe local myelin damage,irregular nuclear neuronal cell morphology,unclear cell membrane boundaries,swelling,deformation,or dissolution of some organelles,glial cells.The morphology is relatively normal,the number is less,the number of synapses decreases,and some synapses are disassembled.In group C,intima thickening,but the state of vascular endothelial cells normal,no inflammatory cell exudation,some areas of severe myelin damage,neuronal nucleus morphology is more regular,cell membrane boundaries clear,organelle swelling,deformation or dissolution of lighter than the B group.The number of astrocytes and microglia cells are incresed,normal morphology,complete structure,good condition,abundant local synapses.On the time point of 7d,in group B,vascular endothelial cell deformation,severe myelin damage,significantly reduced neuronal organelles,astrocyte swelling,necrosis,disintegration,vacuolization,small glial cells swelling and deformation,the number of small glial cells are reduced.The number of synapses decreased significantly,and a large number of synapses disintegrated.In group C,vascular endothelial cells slightly changed,with severe local myelin damage.Neuron organelles were reduced,but the structure was intact compared to thegroup B.The structure of astrocytes and microglia is relatively complete,with a small amount of synaptic disorganization.On the time point of 14d,in group B,Vascular endothelial cells are deformed,the myelin sheath is unclear in texture,the myelin sheath is structurally blurred,the organelles are increased more than before,the nucleus is irregular in shape,the boundary of the cell membrane is unclear,the nuclear membrane shrinks,and a small amount of astrocytes and microglia cells can be seen.Synapse structure is relatively complete,but the synaptic space is significantly wider.In group C,vascular endothelial cells were slightly changed,organelles were more abundant than before,and the myelin sheaths were more clear in texture and myelin sheath structure than in group B.The nucleus morphology was relatively regular and the cell membrane boundaries were clear.The morphology of astrocytes and microglia is intact,the presynaptic membrane and the postsynaptic membrane are intact,and the synaptic space is not significantly broadened.The electron-dense and ribosome aggregation is abundant in the anterior synapse.The group C can relieve the damage caused by ischemia and hypoxia to the ultrastructure of nervous system and reduce the degree of neuron damage.It is obvious that the effect in group C is better than that of group B.3 The effect of synaptic spaceIntra-group comparison:On the time point of 24h,there was no statistically significant different(P>0.05)between the synaptic space in group C and A.However,we find significant different when compared group B,group A and C(P<0.05).On the time point 7d,the synaptic space in group C and B compared with that in group A show statistically significant different(P<0.05).At the same condition,there was significant different between group B and group C(P<0.05).On the time point 14d,the synaptic space in group C compared with that in group A indicate no statistically significant different(P>0.05),while there was significant different when compared group B with group A and C(P<0.05).Inter-group comparison:In group A,there was no significant difference in synaptic space between the different time points of 24h,7d and the 14d(P>0.05).In the group B,the synaptic space at the time points of 14d was wider than the 24h and the 7d(P<0.05);while there was no statistically difference(P>0.05)on the synaptic space between the time points of 7d and 24h.In group C,the synaptic space at the time points of 7d was wider than that at the 24h(P<0.05);but there was no statistically difference(P>0.05)in synaptic space between the different time points of 14d and 24h;the synaptic space at the 14d was shorter between that at the 7d,and there was statistically difference(P<0.05).4 The effect on synaptic vesiclesIntra-group comparison:The number of synaptic vesicles in group C and B compared with that in group A was statistically significant different(P<0.05)on time point of 24h,7d and 14d.There was no significant between group C and group B(P>0.05)on the time points of 24h and 7d.When comes to the time points of 14d,the number of synaptic vesicles in group C was significantly increased compared with group B,and there was significant different between group B and group C(P<0.05).Inter-group comparison:In the three groups,there was no significant difference in synaptic space between the time points of 24h,7d,and 14d(P>0.05).Conclusions1 Cluster needling of scalp acupuncture can significantly improve the neurological score of cerebral ischemia rats,and promote the recovery of motor function of rats.2 Cluster needling of scalp acupuncture can reduce the damage degree of nerve cells and improve the ultrastructure of ischemic penumbra in MCAO rats.3 Cluster needling of scalp acupuncture can increase the number of synaptic vesicles in the ischemic penumbra,effectively protect the structure and function of the synaptic space. |
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