| Objective:(1)To observe the effect of electro-acupuncture(EA)on behavior,brain tissue morphology and expression of Nestin and proliferating cell nuclear antigen(PCNA)in the ischemic lateral subventricular zone(SVZ)of rats with cerebral ischemia.(2)To investigate the effect of EA on blood lipid,behavior,brain tissue morphology.And to explore the effect of EA on expression of Nestin and PCNA in SVZ and ischemia penumbra in hyperlipemia combined with cerebral ischemia rats.Methods:Experiment 1:24 adult male Sprague-Dawley(SD)rats were randomly divided into sham-operation,model,EA1 and EA2 group.Rats in EA1 group were needled at bilateral ST40 with EA(2/100 Hz,1-3 mA),once daily for 7 days.Rats in EA1,EA2 and model group were induced to cerebral ischemia by 50%FeCl3 at day 8.Rats in EA groups were needled at ST40 and GV20 after operation,once daily for 7 days.The behavioral tests were detected by neurological deficit score(NDS)24h post-operation.Hematoxylin and eosin(HE)staining was used to observe the morphological changes of ischemic penumbra.Immunohistochemistry was used to detect the expression of Nestin and PCNA in SVZ.Experiment 2:41 adult male SD rats were randomly divided into normal group and high fat diet(HFD)group.Rats in HFD group were fed with HFD for 42 days to establish hyperlipidemia model,and then randomly divided into sham-operation,model,EA1 and EA2 group.Rats in EA1 group were needled at ST40 with EA(2/100 Hz,1-3 mA),once daily for 7 days.At the 50th day,rats in model and EA1,EA2 group were induced to cerebral ischemia by 50%FeCl3.Rats in EA1,EA2 groups were needled at ST40 and GV20,once daily for 14 days.The behavior was assessed by NDS.Blood lipids were detected by biochemical method.HE staining was used to observe the morphological changes of ischemic penumbra.Immunohistochemistry was used to detect the expression of Nestin and PCNA in SVZ and ischemia penumbra Results:Experiment 1:(1)Behavior tests:Animals exhibited symptoms of reduced diet,weight loss,flexion of the right forelimb,and turning in place in the model group at 24h post-operation.Compared with the model group,the NDS of the EA groups were significantly reduced(P<0.05),and the symptoms of nerve defects were more alleviated.(2)Brain tissue morphology:One day after ischemic,the model group showed a widening of the gap around the neurons.The nerve cells were scattered,the nucleus was condensed,and some blood vessels exuded.After EA treatment,cell vacuole degeneration decreased,and the tissue morphology tended to be normal.(3)Immunohistochemistry:Nestin-positive expression in the sham-operation group was slightly expressed in SVZ.The Nestin area%was increased in the model group at day 1,7 post-operation(P<0.01).Compared with model group,Nestin area%at lateral ventricle extension increased from day 1(EAl P<0.01,EA2 P<0.05)to day 7 post-operation(P<0.01).Nestin area%at lateral wall in the two EA groups increased at day 1 post-operation(P<0.01)and there was not significant difference at day 7 post-operation(P>0.05).Compared with model group,PCNA area%at lateral ventricle extension increased at day 1 post-operation in EA1(P<0.01).There was not significant difference between two EA groups(P>0.05).But there was a tendency that Nestin area%in EA1 group was higher than EA2 group.Experiment 2:(1)Behavior tests:24h after cerebral ischemia,the rats in the model group had higher NDS,exhibited symptoms such as weight loss,flexion of the right forelimb,in suit circling.The NDS of EA groups were significantly lower than that in the model group(P<0.05).The symptoms of neurological deficiency in the EA1 group were more alleviated than that in EA2 group.(2)Blood lipids:The TC,LDL in the model group increased(P<0.01)and HDL decreased(P<0.01)at day 1,7,14 post-operation compared with normal group.Compared with model group,TC and LDL decreased(P<0.05),HDL increased in EA1 group(P<0.01),while HDL increased(P<0.05),LDL decreased(P<0.01)in EA2 group.There wasn't significant different between EAl and EA2 group(P>0.05).(3)Brain tissue morphology:Neurons in the cortical and striatum of the sham-operation group were uniformly distributed and structurally intact,which were basically consistent with the morphology of the normal group.In the model group,the neurons of the ischemic penumbra were necrotic,the nucleus was condensed,a large number of vacuole-like structures were seen.Red blood cells exuded in the blood vessels,and the tissue morphology showed cerebral ischemia as a whole.After EA treatment,the normal neurons increased significantly,and the cell morphology was close to normal.(4)Immunohistochemistry of Nestin:At the lateral ventricle extension of ipsilateral forebrain,the Nestin area%was significantly increased in the model group compared with sham-operation group(P<0.05)at day 1,7 post-operation.Compared with model group,Nestin area%in EA1(P<0.01)and EA2 group(P<0.05)increased at day 1,7,14 post-operation.The Nestin area%in the EA1 group was higher than that of the EA2 group,but there wasn't significant different between two groups(P>0.05).At the lateral wall of the ipsilateral forebrain,Nestin area%was significantly increased in model group at day 1,7,14 post-operation compared with sham-operation(P<0.05).Compared with model group,Nestin area%increased at day 1,7 post-operation in EA1(P<0.05),and increased at day 7 post-operation in EA2(P<0.05).There wasn't significant different between EA1 and EA2 group(P>0.05).At ischemic penumbra,the Nestin area%was increased in the model group at day 1,7,14 post-operation(P<0.01).Compared with model group,Nestin area%increased at day 1,7 post-operation in EA1(P<0.05),while increased at day 14 post-operation in EA1(P<0.05).The Nestin area%in the EA1 group was higher than that of the EA2 group,but there wasn't significant different between two groups(P>0.05).(5)Immunohistochemistry of PCNA:At the lateral ventricle extension of ipsilateral forebrain,the PCNA area%was significantly increased in the model compared with sham-operation at day 1,7 post-operation(P<0.05).Compared with model group,PCNA area%increased at day 1,7,14 post-operation in EA1(P<0.01),while increased at day 14 post-operation in EA2(P<0.01).At the lateral wall of the ipsilateral forebrain,PCNA area%was increased in model group at day 1,7 post-operation(P<0.05).Compared with model group,PCNA area%increased at day 1,7,14 post-operation in EA1(P<0.05),and increased at day 7 post-operation in EA2(P<0.01).At ischemic penumbra,the PCNA area%was increased in the model group at day 1,7,14 post-operation(P<0.01).Compared with model group,PCNA area%in EA1(P<0.01)and EA2 group(P<0.05)increased at day 1,7,14 post-operation.The PCNA area%in the EA1 group was higher than that of the EA2 group,but there wasn't significant different between two groups(P>0.05)Conclusion:(1)EA stimulate ST40+GV20 can improve nerve function,reduce ischemic brain injury,promote the expression of Nestin and PCNA in the SVZ and advance nerve repair after brain injury.EA before cerebral ischemia is more effective(2)EA stimulate ST40+GV20 can regulate blood lipid metabolism,alleviate the symptoms of neurological deficit,improve the morphology of penumbra in rats with hyperlipidemia and cerebral ischemia,EA stimulate ST40 at hyperlipidemia stage and then stimulate ST40+GV20 at post-ischemic is more effective than EA after the onset of ischemic injury.(3)EA may improve the repair and regeneration of the brain by promoting the proliferation of neural stem cells in SVZ on hyperlipidemia-cerebral ischemia injury. |
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