| Obiective:To research the protective mechanism of Qufeng TongQiao Decoction on hippocampus mitochondrial(MIT) in rat with vas cular dementia(VD), by the method of studying on mitochondria (MIT) of VD rats superoxide dismutase(SOD), malondialdehyde(MDA), cytoch-rome oxidase(COX), Cytochrome C Oxidase isoformⅡ(COXⅡ) gene expression and ultrastructural changes in rats of MIT. Methods:1. grouping methods:Using the Morris water maze experiment filter60male SD rats to meet the requirements before and after the modeling, permanent ligation of the bilateral common carotid artery(2-VO) in preparation of VD rat model; which were randomly divided into sham operation group, model group, nimodipine group(referred to as "positive group"), Qufeng Tongqiao Decoc tion high dose group(referred to as""high dose group"), Qufeng Tongqiao Decoction medium dose group(referred to as "medium dose group"), Qufeng Tongqiao Decoction low dose group(referred to as "low dose group"),each group contained10rats;2. Dosiong method:positive group were feed with nimodipine6.25mg/kg/d,Qufeng TongQiao Decoction high、 medium、low dose group were given Qufeng TongQiao Decoction26.8g/Kg/d,13.4g/Kg/d,6.7g/Kg/d, sham operation group and model group were given equal volume of distilled water, orally for30days;3. materials:Using the Morris water maze detect the behavior of each group of rats, randomly selected2rats for cardiac perfusion fixation of each group, fetch brain tissue; the remaining of the rats were taken hippocampus after putting to death, which were made into hippocampus homogenate, differential centrifugation method for extracting MIT, stored at-80℃;4. Target detection:chemical colorimetric determination of rat SOD activity, MDA content, ELISA method for the activity of COX;RT-PCR method for the detection of COXⅡ mRNA expression, light microscope and electron microscope observation on hippocampus cell morphological changes in mitochondria. Results:1. Behavioral learning situation:compared with the sham operation group, model group、positive group and high、medium、low dose group escape latency(EL) was prolonged, platform quadrant dwell time is shortened, the virtual platform to reduce the number of spans, the differences were statistically significant(P<0.05); compared with the model group,positive group、high、medium dose group, the EL is shorted along with the training time is prolonged, platform quadrant dwell time and across the virtual platform number increased significantly, the difference was statistically significant (P<0.05), low dose group EL decreased,but there was no statistical significance(P>0.05), compared with positive group, high、 medium dose group, the EL is shorted, platform quadrant dwell time and across the virtual platform number increased, the difference was statistically significant (P<0.05), compared with low dose group, high、medium dose group, the EL is shorted, platform quadrant dwell time and across the virtual platform number increased, the difference was statistically significant (P<0.05).2.SOD、MDA、COX detection:compared with the sham operation group, model group、positive group and high、medium dose group’s SOD activity decreased, the content of MDA increased, the activity of COX decreased, the difference was statistically significant (P<0.05); compared with the model group, the positive group and high、medium dose group increased the activity of SOD, the content of MDA decreased, the activity of COX increased, the differences were statistically significant(P<0.05); compared with the positive group, high、medium dose group SOD activity was enhanced, the content of MDA decreased, the activity of COX was enhanceed, the difference were statistically significant(P<0.05); compared with the low dose group, high、medium dose group’s activity of SOD increased, content of MDA decreased, the activity of COX increased, the difference was statistically significant(P<0.05).3. light microscopic observation on hippocampus CA1pyramidal cells results:sham operation group of pyramid al cells in layer3～4, neatly arranged closely; integrity of the cells, morpholog ically normal, clear bound-ary, the nucleus is big and round, with abundant cytoplasm; model group pyramidal cell loss significantly, layer of aligned1～2cells, scattered; dividing line is not clear, deformation of a large number of cells;nuclear condensation and fragmentation, dark blue, cytoplasmic conden sation; positive group, high、 medium dose group, pyramidal cell morphology has been improved obviously.4. electron microscope results:sham operation group MIT structurally normal, crest clear structure; model group of MIT marked edema of rupture, MIT crest structure is clear, part MIT shrinkage; positive group, high、medium dose group the damage of MIT are significantly improved; MIT changes in low dose group between positive group and model group.5. COXII mRNA gene expression:All purpose gene band appeared, compared with the sham operation group, model group gene band dimming, each group gene band brightness relatively light; relative gray value:compared with the sham operation group, model group and treatment group relative to gray value is reduced, the difference were statistically significance(P<0.05); compared with the model group, the positive group and high、medium dose group,gray value increase obviously, the difference was statistically significant (P<0.05); compared with the positive group, high、medium dose group of gray value increases, the difference was statistically significant (P <0.05); compared with the low dose group, high、medium dose group gray value is higher, the difference was statistically significant (P<0.05). Conclusion:1. QufengTongqiao Decoction can improve the learning and memory function of VD rats, improve hippocampus pyramidal cells and frontal cortex morphology of MIT VD rat.2. The Protective effect of Qufeng Tongqiao Decoction on VD rat mitochondrial oxidative in jury showed dose-effect relationship.3. Possible mechanism:Through enhanced SOD activity, reduce the MDA contents, exert an antioxidant effect, increased COXⅡ expression and activity of COX, protect the function of MIT, improve cognitive function of VD rats. |
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