Objective:To discuss the mechanism of scalp cluster-needling on learning and memory disturbance big rats induced by chronic alcoholism, by observing the efficiency of learning and memory ability. HIF-1α and VEGF expression. The objective is to provide theoretical basis of treating chronic alcoholism by scalp cluster-needling on clinical.Methods:100wistar big rats were randomly divided into a control group, and an alcohol intragastric group. The chronic alcoholism models were established by intragastric infusion with alcohol for6weeks. Determined the big rats’learning and memory ability by Morris water maze6weeks after alcohol intragastric,and divided successfully modeled rats into a model group, a Aricept group and a scalp cluster-needling group.There was no treatment for control group and model group. The Aricept group were given Aricept0.05mg/kg by gavage,once daily and the treatment last for4weeks.The scalp cluster-needling group were given scalp cluster-needling,acupunctured Baihui and both points2mm aside, remained the needles for30min, once daily and the treatment last for4weeks. Detected the big rats’learning and memory ability by Morris water maze10weeks after alcohol intragastric (4weeks after treatment), observed form changes of hippocampus cells by light microscope, determined protein expression of hippocampus HIF-1α、 VEGF by Immunohistochemistry and expression of hippocampus VEGFmRNA、HIFmRNA were determined by real-time RT-PCR.Results:1. Results of Morris water maze:Learning and memory ability were markedly decreased in the model group.the Aricept group and the scalp cluster-needling group after6weeks alcohol intragastric,with significant differences compared with control group (P<0.05);The learning and memory ability of Aricept group and the scalp cluster-needling group has been increased after4weeks treatment,compared with model group there were significant differences (P<0.01); there were no significant differences between Aricept group and the scalp cluster-needling group (P>0.05).2. Results of hematoxylin-eosin staining:There were no marked pathological changes in control group, while the neuro cells in model group were markedly decreased with cells sparsely arranged, seriously deformed, karyopyknosis, cytoplasm disappeared, uneven dye color marks and fuzzy neucleolus. Neuro cells in Aricept group and the scalp cluster-needling group were regularly arranged and formed, with little pycnosis pyramidal cell. Brain tissue damages were markedly reduced compared with model group.And there are more pyknosis cells in Aricept group compared with the scalp cluster-needling group.3. Results of immunohistochemical staining on HIF-lα、VEGF: Neuro cells in control group were regular with a little positive cells express of HIF-1α、VEGF,the number of neurons and the distribution is normal,closely arranged in an orderly manner,with circular,large,light dyed and clear nucleus; The positive expression of HIF-1α、VEGF in modele group,Aricept group and the scalp cluster-needling group were increased with significant differences compared with control group(P<0.05,P<0.01);There were less positive expressions in Aricept group and the scalp cluster-needling group,compared with model group with significant differences (P<0.05,P<0.01). It is obviously decreased in the scalp cluster-needling group,compared with Aricept group,there is significant differences in positive expression rates (P<0.01).4. Results of HIFmRNA、VEGFmRNA detected by real-time RT-PCR:Expression of THmRNA、VEGF mRNA in control group was exceedingly weak, which were significantly expressed in model group, Aricept group and the scalp cluster-needling group;HIFmRNA、VEGF mRNA were remarkablely expressed in model group, Aricept group and the scalp cluster-needling group,more than the control group;And they were less expressed in Aricept group and the scalp cluster-needling group than the control group.And Aricept group expressed higher than the scalp cluster-needling group.Conclusion:1.Chronic learning and memory disturbance big rats models induced by chronic alcoholism, established by alcohol intragastric, and commented by behaviouristics (Morris water maze) and histology (HE staining), were successful and reliable.2.Scalp cluster-needling can improve chronic alcoholism big rats’learning and memory ability by improving pathological changes of hippocampus tissue, inhibiting the neuronal apoptosis, reducing the pathological changes of nerve cells.3.Scalp cluster-needling can decrease expression of HIF-1α and VEGF, cut HIF-1α mRNA and its target gene expression of VEGF mRNA and HIFmRNA, reduce neurons in the brain damage after brain injury, and also reduce neuronal apoptosis in order to play a protective function.4.Scalp cluster-needling can be used on pretreatment of learning and memorizing disorder caused by chronic alcoholism,and is expected to be one of the new therapies. |