| Cerebral ischemia is a common acute cerebrovascular disesase. The key of treatment is to save the dying neurons in peri-infarct region and to promote rehabilitation of neuronal function. Acupuncture on nuchal region (ANR) exerts a remarkable curative effect on acute stage and sequela stage of cerebral ischemia reperfusions (CIR), but the mechanism research of acupuncture on nuchal region is just at the beginning. This project is to proceed with how the elctroacupuncture on nuchal region (EANR) intervene the endogenesis neural stem cell. To investigate the mechanism of this action will pioneer a new realm of science research.Objective: To study the expression of nerve cell proliferation associated factors following CIR in rats and the influence of EANR compared with ANR. Try to reveal the mechanism of two therapies for CIR.Methods: Focal cerebral ischemia was induced by intraluminal middle cerebral artery occlusion (MCAO) method using a monofilament thread in rats. Male Wistar rats were divided randomly into four groups: sham group, model group, EANR group and ANR group; each group was separated into 1d, 3d, 7d, 14d after reperfusion as subgroup for investigation. (1) Neurological deficits and infarct volumes were observed in rats; (2) HE staining and electron microscope were used to look into the changes of histomorphology; (3) The expression of NF-κBp65,TGF-β1,NCAM and Nestin protein were measured by immunohistochemistry technique respectively and how EANR and ANR influence the expression was observed; (4) The expression of TGF-β1,PSA-NCAM,Nestin and SCF mRNA were measured by reverse transcription-polymerase chain reaction and how EANR and ANR influence the expression was investigated.Results: (1) Neurological deficit scores in EANR and ANR therapy groups were significantly higher than the ones in the model group and the infarct volume was significantly lower than the ones in the model group (p<0.01, p<0.05). And there was a significant difference between the EANR and ANR group (p<0.05). (2) The results of HE staining showed that after acupuncture there is light neural cell swelling but more neurons and newborn blood capillaries compared with the model group. Electron microscope results indicated that the cerebral ultrastructure was significantly meliorated in the EANR and ANR group. There were plenty of golgicomplexes in neuron, the structure of mitochondrion and other cell organs were full and there were more synaptic vesicles in the EANR and ANR group rats. Furthermore, the morphological change in EANR group was much better than the ones in the ANR group. (3) Immunohistochemistry results indicated that compared with the model group, EANR and ANR could obviously increase the expression of TGF-β1,NCAM and Nestin protein in cerebral cortex and striatum while they could inhibit the expression of NF-KBp65 in the early stage after ischemia reperfusion. There was a significantly difference between EANR and ANR group (p<0.01 or p<0.05). (4) RT-PCR results pointed out that compared with the model group, EANR and ANR can increase the expression of TGF-β1, NCAM, SCF and Nestin at the different time point distinctly (p<0.01). And there was a significantly difference between the EANR and ANR group (p<0.01).Conclusion: (1) EANR and ANR can save the neurocytes in peri-infarct region and reinforce the blood supply so as to amend neural function and reduce the infarction ratios. (2) EANR and ANR can inhibit the expression of NF-κBp65 in the early stage of CIR and increase the expression of TGF-β1 protein which can inhibit cell apoptosis efficiently. (3) EANR and ANR can increase the expression of TGF-β1, NCAM and Nestin protein which can induce blood vessel regeneration and the proliferation, differentiation of neural stem cell so as to promote rehabilitation of neuronal function after focal ischemia and alleviate the injury induced by CIR. (4) EANR and ANR can improve the expression of TGF-β1, NCAM, Nestin and SCF mRNA which can accelerate the proliferation, migration and differentiation of neural stem cell so as to promote neurogenesis, reparation and reconstitution of neural function. (5) Through comparison study we find that the EANR acts better than the ANR in the aspects hereinbefore which may be related to different stimulus intensity and the effect of electric field.
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