| Objective: We observed the curative effect of electropuncture (2Hz,80Hz, and2/80Hz, variable frequency) on vascular dementia (VascularDementia, VD) mice to compare cupping treatment for frequency, discuss therole of VD mechanism, and provide the scientific VD electroacupunctureparameters to clinical treatment of acupuncture.Methods:1The effect of electropuncture with different frequency on Praxiology andPathomorphology on Hippocampus CA1region of VD miceThe Kunming mice were randomly divided into six groups, the shamoperation group, the model group, the nimodipine group, the2Hzelectropuncture groups, the80Hz electropuncture groups, the2/80Hzelectropuncture groups. VD mice were established by blocking the blood, tailbleeded and repeated cerebral ischemia-reperfusion, while the sham groupwas only a process. The blood wasn't blocked and the tail wasn't bleeded.They were treated on the4d. The baihui, dazhui, gushu, zusanli were choosedon the electropuncture group, acupuncted with filifo, connected withelectroacupunctrue instrument, continuous wave,2times/s,80times/s anddensity wave2/80Hz. Ten minutes every time and one time a day. Nimodipine30mg/kg/d was given to the Nimodipine group by watering stomach. After15d treatment, we examined the study score by water-maze experiment andjumping stand experiment, and the memory score on the next day. Then, themice were beheaded and the left brains were taken out on the ice plate, thetissue from optic chiasm to nipple was taken and fixated in4%formal dehyde.The tissue was stained by HE (Hematoxylin and Eosin). We observedmorphologic changes of hippocampus CA1region under light microscope. 2The effect of electropuncture with different frequency on CGRP, NOS,oxygen free radicals and TchE in hippocampus of VD miceThe dividing groups, establishing models and treating methods were allsame to1. The mice were beheaded, the right hippocampus was make into10%homogenate. The homogenate was centrifugated for20min at4℃,4000r/min, the supernatant was extracted. Then, we used biuret method todetermin protein level, the radiation immune analysis in the hippocampusCGRP content determination of organization, spectrophotometry NOS, TchEactivity and xanthine oxidase method to detect the activity changes of SOD,thio-barbituric acid compared color analytical method to detect the MDAcontent.3The effect of electropuncture with different frequency about caspase-3(pro-apoptotic protease) positive expression and the mean optical densityvalues on Hippocampus CA1region of VD miceThe dividing groups, establishing models and treating methods were allsame to part1. After the examination of Praxiology, the mice were beheadedand the left brains were taken out on the ice plate, the tissue from optic chiasmto nipple was taken and fixated in4%formal dehyde, covered with paraffin,make into coronary slice (5um). Three randomly selected,immunohistochemical SABC method was used to detect positive expressionof caspase-3in hippocampal CA1region and observe the different frequenciesof electroacupuncture on the nerve cells of the hippocampal CA1region ofpro-apoptotic protease positive expression and the mean optical density value.Statistical analysis of all data was conducted with SPSS.Results:1The effect of electropuncture with different frequency on Praxiology andCA1pathomorphology of VD mice1.1Results of praxiology experimentResults of jumping stand experiment: Compared with sham operationgroup, the react time of the model group was extended, latent period wasshorten and the mistake number of times was significantly increased (P<0.01). Compared with model group, both EA and nimodipine treatmentcan shorten the reaction time, extend the incubation period, reduce themistake number of times (P<0.05or P<0.01). The2/80Hz group was betterthan others (P<0.01). There was no significant difference among2Hz,80Hzand2/80Hz group (P>0.05).Results of water-maze experiment: Compared with sham operation group,the swimming time of the whole line of the model group was obviouslyextended and the mistake number of times was significantly increased(P<0.01). Compared with model group, both EA and nimodipine treatment candecrease the swimming time of the whole line and the mistake number oftimes (P<0.05or P<0.01). Among them, the EA2Hz group2/80Hz groupwere better than nimodipine group and80Hz group (P<0.05or P<0.01).Group2Hz and2/80Hz group were no difference (P>0.05). Nimodipine and80Hz group were no difference (P>0.05).1.2The effect of electropuncture with different frequency pathomorphologyon Hippocampus CA1region of VD miceThe hippocampal CA1region was stained by HE, the generalmorphological observation showed that: Model group the hippocampalpyramid cells in CA1area were evidently lost and arranging disorderly, shapeowe rules, the nucleus quality uneven, nuclear become pyknotic serious. Thetreatment groups were improved, nimodipine group and80Hz group, cellsreduce loss, the arrangement was still owe neat, cell approximate circle andthe nuclei were uniform quality.2Hz group of cells tightly packed packagewas not very regular, nearly circular cell, the nucleus mass was more uniform.2/80Hz group, only see a small number of cells depigmentation arrangedcompactly structured cell round, clear nuclei, nuclear quality uniform.2CGRP,NOS,oxygen free radicals and TchE in hippocampus of VD miceResults manifested that: The content of CGRP in model group wasreduced significantly than that in sham operation group (P<0.01). And intreatment groups, the content of CGRP was increased than model group(P<0.05or P<0.01). Among them, the EA2Hz group2/80Hz group were better than nimodipine group and80Hz group (P<0.05or P<0.01). The2/80Hz group was more prominent than the2Hz group (P<0.05). Nimodipineand80Hz group were no difference (P>0.05).The activity of NOS of model group was raised significantly than thatin sham operation group (P<0.01). Compared with the model group, EA andnimodipine treatment may reduce the activity of NOS (P<0.01). The2/80Hzgroup was better than others (P<0.01). There was no significant differenceamong2Hz,80Hz and2/80Hz group (P>0.05).The content of MDA was raised and the activity of SOD wassignificantly reduced than that in sham operation group (P<0.01). Comparedwith the model group, the treatment groups can reduce the content of MDA,increase SOD activity (P<0.05or P<0.01). Among them, the EA2Hz groupand2/80Hz group were better than nimodipine group and80Hz group (P<0.05or P<0.01). The2/80Hz group was more prominent than the2Hz group(P<0.05). Nimodipine and80Hz group were no difference (P>0.05).The activity of TchE of model group was significantly raised than thatin sham operation group (P<0.01). Compared with the model group, EA andnimodipine treatment may reduce the activity of TchE (P<0.05or P<0.01).Among them,2/80Hz group was better than others (P<0.05). There was nosignificant difference among nimodipine,2Hz and80Hz group (P>0.05).3The effect of electropuncture with different frequency about caspase-3(pro-apoptotic protease) positive expression and the mean optical densityvalues on Hippocampus CA1region of VD miceIn the model group, Casepase-3positive expression and the averageoptical density values were strongly increased (P<0.01). Compared with themodel group, EA and nimodipine treatment can weaken the positiveexpression and reduce the average optical density values (P<0.01). Amongthem, the EA2Hz group and2/80Hz group were better than nimodipine groupand80Hz group (P<0.05or P<0.01). The2/80Hz group was more prominentthan the2Hz group (P<0.05). Nimodipine and80Hz group were no difference(P>0.05). Conclusion: The experiment demonstrated that electroacupuncture withdifferent frequency (acupuncted the points of "Baihui","Dazhui""Geshu", and"Zusanli ") had therapeutical effect on VD mice. Its mechanism mayassociated with increased level of CGRP and activity of SOD, decreasedcontent of MDA and activity of NOS with TchE, thereby, improved bloodcirculation disorders which caused by ischemia-reperfusion in the brain, blockthe neurotoxic cascade reaction against lipid peroxidation damage, adjust thecholinergic nervous system functions, and lower the expressions ofCasepase-3, inhibit the apoptosis of the CA1region of hippocampus nervecells, reduce the damage, therefore, it can improve the study memory obstacleinduced by ischemia-reperfusion in brain. And the treatment by2/80Hz wasmost effective for the VD model mice. |
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