The Effect Of Huayu Kaiyu On The Neural Stem Cells Of Rats With Hyperlipemia Combined With Cerebral Ischemia

Objective To explore the effect and mechanism of "Huatankaiqiao" electro-acupuncture(EA)on blood lipid,neurological function,cerebral infarct volume,and the express of proliferation cell nuclear antigen(PCNA)in ischemic penumbra(IPR)region and sub-ventricular zone(SVZ)region.Methods Seventy-eight SD rats were randomly divided into normal control group,hyperlipidemia combined with sham operation group,hyperlipidemia combined with cerebral ischemia model group,hyperlipidemia with cerebral ischemia EA1 and 2 groups.In addition to the normal group,the rats in the other groups were fed with high-fat diet for 6 weeks.Rats in EA1 group were needled "fenglong"(ST40)once daily for 7 days.Rats in the model group,EA1,and 2 group were established hyperlipidemia with cerebral ischemia model induced by 50%FeC13.The rats in EA group were needled GV20 and ST40,once daily for 14 days.The blood lipids were detected by biochemical method,and the behaviour were evaluated by neurological scores.The infarct volume was detected by 2,3,5-Triphenyltetrazolium chloride(TTC)method and Brain neuronal morphology was observed by hematoxylin-eosin(HE)staining.The pathological morphology was studied with a light microscope and the expression of PCNA was detected by immunohistochemistry.Results(1)Lipids:-There were significant differences at the time points.TG and LDL had significant different at each time points during the treatment of hyperlipidemia-cerebral ischemia by sphericity tests(P<0.01).Compared with the normal group,the TC and LDL-C levels in the model group increased(P<0.05,P<0.01),and the HDL-C decreased(P<0.05,P<0.01).The model of the high-lipid model was established successfully.On the 7th and 14th days after surgery,TG in the model group increased significantly(P<0.01).After acupuncture treatment,compared with the model group,the TC,TGand LDL were decreased in EA1,2 groups on 7th days postoperation(P<0.01),while HDL increased.At 14 th days after surgery,TC,TG,and LDL decreased in EA1,2.groups on 14th days postoperation,while HDL increased.(2)Neuroethology:The rats in the model group exhibited symptoms such as forelimb flexion,ipsilateral hindlimb weakness,in situ circling and other symptoms at the 24h postoperation and the neurological deficit scores were higher.The neurological deficits in the rats in EA1,2 groups were significantly lower than those in the model group(P<0.01).There was a significant difference between two EA groups.(3)Cerebral infarct volume:the cerebral infarction volume in EA groups was significantly bigger than that in the model group(P<0.01).(4)Brain morphology:In the sham operation group,the cortical nerve cells and glial cells were clearly arranged,the nucleolus was clear,and the cytoplasm was not stained red.One day after cerebral ischemia,the ischemic lesions appeared in the left frontal lobe.After the operation,the model group showed a widening of the gap around the neurons,disordered arrangement of cells,and infiltration of inflammatory cells in some of the blood vessels;necrotic cells increased significantly after 7 days of ischemia.Deep nuclear staining and vacuolization appear.after 14 days of ischemia.The area of necrosis continued to increase.The nuclei of large area cells disappeared,and the interstitial cells loosely showed a fishnet shape.After acupuncture treatment,edema around the ischemic region in EA group was relieved,and the cells also gradually returned to normal.(5)Immunohistochemistry:in the penumbra of the ischemic penumbra and the lateral cerebral ventricle of the ischemic side,compared with the sham group,the positive expression of PCNA was increased and the number of proliferating cells was increased in IPR and SVZ region in the model group(P<0.05).Compared with the model group,the positive expression of PCNA in the EA groups was significantly increased,and the number of positive cells increased.It has significant difference(P<0.05).Compared with EA2 group,it has a rising tendency in EA1 group.Except for the sham group,the positive expression of PCNA in each group began to gradually increase on the 1st day after operation,reached the peak on the 7th day after surgery,and gradually decreased after 14 days.The results suggest that electro-acupuncture acupoints treatment can make the neurons proliferate in IPR and SVZ region of the lateral ventricle,Pre-acupuncture treatment can promote the proliferation of nerve cells.Conclusion "Huatankaiqiao" electroacupuncture can reduce blood lipids,After cerebral ischemia,electroacupuncture can effectively improve the neurological deficit state of rats,reduce the infarct focus,promote the regeneration and proliferation of neural cells in the ischemic penumbra and SVZ area,and improve the pathological morphology of brain cells.The effect is best at 7th days postoperation.Pre-acupuncture treatment for hyperlipemia can play a better therapeutic effect.It can provide a theoretical basis for the stroke treatment in clinic.