| Objective(1)To investigate the effect of "Huatan Kaiqiao" electro-acupuncture(EA)on the effects of GFAP and NGF protein expression in the ischemic lateral subventricular zone(SVZ)in hyperlipemia combined with cerebral ischemia rats.(2)To investigate the effect of EA on behavior,cerebral infarction volume and expression of GFAP and NGF protein in SVZ in ischemic ratsMethodsExperiment 1:90 male Sprague-Dawley(SD)rats were randomly divided into normal,hyperlipemia combined with sham-operation(sham-operation),hyperlipemia combined with cerebral ischemia model(model),EA1and 2 groups.Except for normal group,rats in the other groups were fed with high-fat diet for 42 days.Then,rats in EA1 group were treated with EA at bilateral ST40,once a day for 7 days.On the 50th day,rats in the model and EA groups were established hyperlipemia combined with middle cerebral artery thrombosis model induced by 50%FeC13.Rats in EA groups were treated with EA at ST40 and GV20 points,once a day for 14 days.Behavioral changes were assessed by using neurological deficient score(NDS).Cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL)and low density lipoprotein(LDL)levels were measured.Morphological change was observed by Hematoxylin-eosin staining(HE).The content of GFAP and NGF protein in SVZ region was detected by immunohistochemistry.Experiment 2:72 male SD rats were randomly divided into sham-operation,model,EA1 and EA2.Rats were fed with ordinary food.Rats in EA1 group was treated with EA at bilateral ST40,once a day for 7 days.On the 8th day,the middle cerebral artery thrombosis occlusion model was induced by 50%FeC13.Rats in EA groups were needled GV20 and ST40 points with EA,once a day for 7 days.Neurological behaviors were observed 24 hours after surgery by NDS.2,3,5-triphenyltetrazolium chloride(TTC)staining was used to detect cerebral infarction volume.The expression of GFAP and NGF protein in the SVZ region was observed by immunohistochemistry.ResultsExperiment 1:(1)Neural behavior:In the model group,the NDS was higher,and there were symptoms of ischemic lateral forelimb flexion,hind limb weakness,and rotation to the healthy side.The symptoms of neurological deficient in the EA groups were significantly lighter than that in the model group(P<0.05).Compared with the EA2 group,the symptoms of neurological deficiency in the EA1group were alleviated.(2)Blood lipids:TC and LDL increased significantly(P<0.01),and HDL reduced significantly(P<0.05)in model group after rats were fed with high-fat diet.Compared with the model group,TG reduction in the EA groups was statistically significant at 7d after surgery(P<0.05).TG reduction in the EA1 group was bigger than that of the EA2 group.It indicates that EA pretreatment had better effect on adjusting blood lipids.(3)Brain tissue morphology:The morphology of neurons in the normal group was almost normal.The overall tissue structure was normal.There were many shrinking neurons,and the overall histomorphological structure showed cerebral ischemia in model group.After EA treatment,the edema around the ischemic area was alleviated,the nucleus gradually recovered,and the overall tissue morphology gradually returned to normal.(4)Immunohistochemistry:At the lateral ventricle extension of ipsilateral forebrain,the GFAP area%was significantly increased in the model compared with sham-operation(P<0.05)at Id after surgery.Compared with the model,the GFAP area%in the EA group increased from 1d to 14d after surgery.Compared with the model,the GFAP area%in the EA1 group had statistically significant different at 7d after operation(P<0.05).The GFAP area%in the EA1 group was higher than that of the EA2 group.At the lateral wall of the ipsilateral forebrain,the GFAP area%in model was significantly increased compared with the sham-operation at 1d after surgery(P<0.01).Compared with the model,the GFAP area%of EA1group was statistically significant at 7d after surgery(P<0.05).Compared with the model group,the GFAP area%of the EA1 group increased significantly at 14d after operation(P<0.01);The different in the EA2group with model was statistically significant(P<0.05).Compared with the EA1 group,the GFAP area%in the EA2group was statistically significant(P<0.01).It was higher in the EA1group than that in the EA2group.At the ischemic penumbra,compared with the sham-operation the GFAP area%increased significantly in the model at 1d after surgery(P<0.01).Compared with the model,there was statistically significant different in EA1 group at 7d after the surgery(P<0.05).Compared with the model,there was statistically significant different in EA groups at 14d after the surgery(P<0.01).Compared with the EA1group,there was statistically significant different in EA2 group(P<0.01).At the lateral ventricle extension of ipsilateral forebrain,compared with the sham-operation,the NGF area%in the model was significantly increased at 1d after surgery.Compared with the model,the NGF area%in the EA groups was increased at 7d after surgery.The difference was statistically significant(P<0.01).The NGF area%in the EA1group was higher than that of the EA2group,which had statistically significant difference(P<0.05).At the lateral wall of the ipsilateral forebrain,compared with the sham-operation,the expression of NGF area%rose in the modelat 1d after operation,and the difference was statistically significant(P<0.05).Compared with the model,the NGF area%increased significantly in the EA groups at 7d after operation.Compared with the model,the NGF area%of the EA groups continued to increase,and the NGF area%of the EA1 group was higher than that of the EA2group at 14d after operation.At the ischemic penumbra,compared with the sham-operation,the NGF area%in the model increased significantly(P<0.05)at 1d after surgery.Compared with the model,the NGF area%in the EA1group increased significantly at 14d after surgery(P<0.01).Experiment 2:(1)Neurobehavior:NDS in the model were significantly higher than that in the EA groups(P<0.05).It was shown that ischemia caused severe loss of brain function.EA can improve the recovery of behavior function after cerebral ischemia.(2)Cerebral infarction volume:Compared with the model,the infarct volume in the EA groups decreased.There was significantly difference between model and EA1 group(P<0.05).It showed that EA can reduce the cerebral infarction volume.It was lowest in EA1 group.(3)Immunohistochemistry:GFAP protein was slightly expressed in the lateral ventricle extension and lateral wall of ipsilateral forebrain in the sham-operation.Compared with the sham-operation,it increased significantly in the model(P<0.05).The expression in the EA groups was significantly higher than that in the model and continued until 7 days after ischemia.The NGF area%in the sham-operation was extremely low.It increased from 1d after ischemia.It was highest at 7d after surgery in the EA1 group.It increased highly in sham-operation and EA2 group.It was lowest in the cerebral ischemia group.Conclusion "Huatan kaiqiao" refers to use EA at ST40 point in the stage of hyperlipidemia,and EA at ST40 and GV20 points after hyperlipemia combined with cerebral ischemia.It can regulate blood lipid metabolism,promote the recovery of limb function in cerebral ischemia rats,and reduce the volume of cerebral infarction.Laboratory studies have shown that it can improve the pathological changes of brain tissue,up-regulate the GFAP and NGF proteins in the SVZ region of ischemic rats,and promote the activation of astrocytes and the expression of nerve growth factor.It can provide a theoretical basis for clinical acupuncture treatment of stroke for restoring damaged nervous system. |
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