| Objective: To observed the intervention effect of electropuncture(2Hz, 100 Hz, and 2/100 Hz, variable frequency) on behavior, brain tissue pathological morphology of hippocampal CA1 area, P38 MAPK pathway and Cell Apoptosis of vascular dementia(Vascular Dementia, VD) rats, discuss therole of VD Mechanism, and provide the scientific VD electroacupuncture parameters to clinical treatment of acupuncture.Methods:Screened by Morris water maze system normal rats learning and memory function, it were randomly divided into the sham operation group and the model group. The bilateral common carotid artery repeatedly by intraperitoneal injection of ischeia-reperfusion plus sodium nitrate tome, method of preparation of VD model separation control rats common carotid artery, not injection, sodium nitrate, don,t block the blood flow. Blocking carotid artery after 1min to observe rat is reflection and iris changes to ensure that the brain ischemia. Postoperative survival model rats were randomly divided into the model group, the 2Hz electric acupuncture group, the 100 Hz electric acupuncture group, the 2/100 Hz electric acupuncture group. They were treated on the 3d. The dazhui, baihui, housanli(double), gushu(double), were choosed on the electropuncture group, 0.30×25mm acupuncted with filifo, connected with the Korean electroacupunctrue instrument, respectively with 2 times/s of low frequency, 100 times/s of high frequency and frequency conversion of 2/100 times/s, ten minutes every time and one time a day, continuous treatment of 15 d. Control group and model group are in the same position fixed 10 min. On the 1d and the 17 d after modeling stoke index score and neural disorder score. On the 17 d, we examined water-maze experiment. Then, the rats were beheaded and the left brains were taken out on the ice plate, and the left side of the braintissue was taken and fixated in 4% formal dehyde. The tissue was stained by HE(Hematoxylin and Eosin). We observedmorphologic changes of hippocampus CA1 region under light microscope. The mice were beheaded, the right hippocampus was make into 10% homogenate,conventional paraffin embedding,sectioning undertest.The homogenate was centrifugated for 20 min at 4℃,4000r/min,the supernatant was extracted. Then, we testing the expression of p38 MAPK by immunohistochemistry, the expression of Bax, Bcl-2, Caspase-3 Western Blot, observed cell apoptosis by TUNEL.Results: 1 The effect of electropuncture with different frequency on Praxiology and Pathomorphology on Hippocampus CA1 region of VD rats 1.1 Praxiology scoreThe stoke index score and neural disorder in the electropuncture group and the model group were obvious high than sham operation group on the 1d after modeling. There was no significant difference among electropuncture group, model group(P>0.05). On the 17 d after modeling, the electroacupuncture group stoke index score and neural disorder score compared with the model group was significantly decreased(P<0.01). There were no significant difference among 2Hz group, 100 Hz group and 2/100 Hz group(P>0.05). 1.2 Learning and memory performance testsThe results of water-maze experimentshow:compared with the sham operation group,the model group rats,escape latency significantly prolong and through the virtual platform number decreased significantly, with significant statistical differences(P<0.01). The three kinds of frequency electric acupuncture can shorten the escape latency and to increase the number through the virtual platform(P<0.01 or P<0.05). There were no significant difference among 2Hz group, 100 Hz group and 2/100 Hz group(P>0.05). 1.3 The effect of electropuncture with different frequency pathomorphology on Hippocampus CA1 region of VD ratsThe Model group show lots of cells appear edema, loosely arranged, disorder, cell falls off, the shape of owe rules, the nucleus significantly shrink, deformation, nucleoli and polygon fragments, with nuclear mass unevenly; Each electroacupuncture group pathological changes were improved significantly. compared with model group, cell edema, relieve, loss reduction, relatively tight, neat, cells are round, the nucleus quality clear. 2 The effect of electropuncture with different frequency on the expressions of p38 MAPK, Bax, Bcl-2, Caspase-3 and cell apoptosis in hippocampus CA1 regionof VD ratsCompared with the sham operation group, P38 MAPK positive expression and the average optical density values in the model group, were strongly increased(P<0.01). The three kinds of frequency electric acupuncture can reduce P38 MAPK positive expression and the average optical density values(P<0.01). There were no differences among 2Hz group, 100 Hz group and 2/100 Hz group(P>0.05).Compared with the sham-operation group, the expression level of Bcl-2 protein in the model group decreased significantly and theexpression level of Bax protein increased significantly, with significant differences(P<0.01). Diffrerent frequency electric acupuncture can increase the Bcl-2 protein expression, reduce Bax protein expression,compared with model group significantly statistical differences(P<0.01 or P<0.05). 2Hz, 2/100 Hz group to reduce the Bcl-2, Bax protein expression effect is more significant, compared with 100 Hz group was statistically significant(P<0.01). 2Hz group compared with 2/100 Hz group therapy effect is more prominent(P<0.05).Compared with the sham-operation group, the expression level of Caspase-3 protein in the model group were significantly higher(P<0.01). The Caspase-3 protein expression of electric acupuncture group were significantly lower, significant difference compared with model group(P<0.01); 2Hz, 2/100 Hz group reduce Caspase-3 protein expression is more apparent, compared with 100 Hz group was statistically significant(P<0.01); 2/100 Hz group compared with group 2Hz treatment effect is more significant(P<0.01).Model group increased the average optical density value increases, significant difference compared with control group(P<0.01); The average optical density value is reduced, has the remarkable difference compared with model group(P<0.01); There were no significant difference among groups(P> 0.05).Conclusion: 2Hz and 100 Hz,2/100 Hz frequency electroacupunctures timulation at "dazhui", "baihui", "housanli", "geshu" can relieve cerebral ischemia reperfusion of VD model rats hippocampal nerve cells caused by pathological damage, improve the cognitive impairment of VD model rats. The mechanism may be through the influence of VD model rats hippocampal neurons P38 MAPK, Bcl-2, Bax, Caspase 3 mediated apoptosis pathway, inhibition of VD the excessive apoptosis of hippocampus neural cells of rats, reduce secondary nerve injury,repair damaged neurons. |
PDF Full Text Request