Role Of CREB Phosphorylation In Neurogenesis In Vivo Activated By Electro-acupuncture Combined With Repetitive Transcranial Magnetic Stimulation After Focal Cerebral Ischemia

PartⅠStudy on the role and mechanism of PKA-CREB SignalTransduction System in neurogenesis activated by electro-acupuncturecombined with repetitive Transcranial Magnetic Stimulation after focalcerebral ischemia in adult ratsObjective:To investigate the role and mechanism of PKA-CREB Signal Transduction System inneurogenesis activated by electro-acupuncture combined with rTMS after focal cerebralischemia in adult rats.Methods:The model of transient focal ischemia was made by the middle cerebral arteryocclusion.One hundred and fifty Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA combined with rTMS group.To observe theneurologic impairment rating,ability of learning and memory at the 7th,14th and 28th dayafter infarction respectively.Meanwhile,Immunohistochemistry and Western blot wereconducted to detect the expression of PKA and pCREB in hippocampus of ipsilateralhemispheres.Results:(1) The improvement of neural motor function deficits as well as the indexes oflearning and memory were more obvious in EA group,rTMS,EA combined with rTMS group compared with the model group (p＜0.01,p＜0.05),and the improvement were themost obvious in the EA combined with rTMS group.(2) The number of positive cells and the average gray density of PKA and pCREBexpression in hippocampus of ipsilateral hemispheres in the model group were higher at the7th day,lower at the 28th day than that of nomal group (p＜0.05),higher in EA group,rTMS,EA combined with rTMS group than that of model group in every time point andnomal group at 7th and 14th day (p＜0.05),higher in EA combined with rTMS group thanthat in EA group,rTMS group at 7th and 14th day,and there was no difference between EAgroup and rTMS group.(3) Positive correlation existed between pCREB positive expression with NSCsproliferation.Conclusion:EA combined with rTMS can conspicuously promote the functional recovery aftercerebral arterial thrombosis,upregulate the expression of related signaling molecule ofPKA-CREB transduction system in hippocampus of ipsilateral hemispheres,which mightbe one of the important mechanisms of EA combined with rTMS in treating ischemic braininjury.PartⅡEffects of electro-acupuncture combined with repetitiveTranscranial Magnetic Stimulation on migration and differentiation ofinherent neural stem cells after focal cerebral ischemia in adult ratsObjective:To investigate the effects of electro-acupuncture(EA) combined with repetitive Transcranial Magnetic Stimulation (rTMS) on migration and differentiation of inherentneural stem cells after focal cerebral ischemia in adult rats and to explore the mechanism ofEA combined with rTMS in treating ischemic brain injury.Methods:The model of transient focal ischemia was made by the middle cerebral arteryocclusion.One hundred and twenty Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA combined with rTMS group.To observe theexpression of polysiolylated neural cell adhesion molecule(PSA-NCAM) andPSA-NCAM/5-bromodeoxyuridine (BrdU),NeuN and NeuN/BrdU,Calbindin in differentencephalic regions of ipsilateral hemispheres by immunohistochemical andimmunofluorescence staining at the 7th,14th and 28th day after infarction respectively.Results:(1) The number of PSA-NCAM-positive cells and PSA-NCAM/BrdU positive cells indifferent encephalic regions of ipsilateral hemispheres increased at day 7after MCAO(p＜0.05),reached maximum at day 14 (p＜0.05),and decreased markedly at day 28,but itwas still elevated compared with that in normal group.There were significant differences inEA group,rTMS group and EA combined with rTMS group when compared with that ofmodel group at day 7 and 14 (P＜0.05).There were significant differences in EA combinedwith rTMS group when compared with that of EA group and rTMS group at day 7 and 14(P＜0.05).However,there were no statistic differences in EA group,rTMS group and EAcombined with rTMS group when compared with that of model group at day 28(P＞0.05).(2) The number of NeuN-positive cells and NeuN/BrdU positive cells in hippocampusincreased at day 7after MCAO (p＜0.05),reached maximum at day 14 (p＜0.05),anddecreased markedly at day 28,but it was still elevated compared with that of normalgroup.There were significant differences in EA group,rTMS group and EA combined withrTMS group when compared with that of model group at day 7 and 14 (P＜0.05).Therewere significant differences in EA combined with rTMS group when compared with that of EA group and rTMS group at day 7 and 14 (P＜0.05).However,there were no statisticdifferences in EA group,rTMS group and EA combined with rTMS group when comparedwith that of model group at day 28(P＞0.05).(3) In EA combined with rTMS group,the neuron arrangement in peripheral cortex ofinfarction was regular just with mild cell loss and some darken dyeing nucleus.The averageoptical density of Calbindin in EA combined with rTMS group were higher than that inmodel group,EA group and rTMS group (P＜0.05).Conclusion:EA combined with rTMS can promote the proliferation,migration and differentiationof inherent neural stem cells in different encephalic regions of ipsilateral hemispheres,promote the neural regeneration and neurofunctional restoration,which might be one of theimportant mechanisms of EA combined with rTMS in treating ischemic brain injury.