| Objective: To explore the role of Yishenjiannao prescription on oxidat ive stress injury in rats with vascular dementia and effects on the ex pression of NF-E2-related factor 2(Nrf2)and Glutathione-dependent an tioxidant enzyme glutathione peroxidase 4(GPX4)in hippocampal tissu es.Methods:Taking the relationship between related factor Nrf2,GPX4 protein expression and oxidative stress as the research point,65 male SD rats in a SPF grade were selected,after 3-day's adaptive breeding,according to the random and contrast principles,6 rats were divided into the sham operation group,the remaining 59 rats only adopted the animal model of Vascular dementia established by improved 2-VO method,screened with Morris water maze experiment four weeks later,rats with successful modeling were randomly divided into model group,positive control group and Yishenjiannao group.Positive control group and Yishenjiannao group were lavaged with regulated dose,while the sham operation group and model group were treated with the same volume distilled water.Four weeks after gavage,Morris water maze experiment was performed to compare the behavioral changes of rats in each group.Then the rats were sacrificed,and the left cerebral cortex and hippocampus were isolated from their brains.The pathological morphology of the hippocampal CA1 region of the rats with vascular dementia was detected by Hematoxylin-Eosin(HE)staining.The right cerebral cortex and hippocampus were isolated and the expression of Nrf2 and GPX4 proteins was detected by Western blot.The experimental data were processed by SPSS23.0 statistical software and Graphpad prism7.0mapping software.The measurement data were expressed as mean ± standard deviation((?)±s),and the samples of each group were consistent with normal distribution and homogeneity of variance.One-way ANOVA was used for inter-group comparison,and LSD methodwas used for multiple comparison.Those with unequal variances were compared by tam-black mud method.with statistically significant difference(P<0.05 or P<0.01).Results: 1.Morris water maze detection: in the positioning sea trial,the escape latency period of rats in each group was shortened at different degree as the training time extended.Two days before the experiment,there was no significant difference in EL in each group(P>0.05).On the third day,the average EL of rats in the model group was significantly longer than that in the sham operation group(P<0.01).Compared with the model group,rats in the Yishenjiainao group and the positive control group had a tendency of EL shortening,but there was no statistical significance(P>0.05).On day 4 and 5,compared with the model rats,the average EL of each group was significantly shortened(P<0.01).In the space exploration experiment,compared with the sham operation group,times of passing through the original platform and the residence time in the target quadrant of the model group were all reduced(P<0.05).Compared with the model group,times of passing through the original platform and the residence time of the target quadrant were significantly increased in the Yishenjiannao group and the positive control group(P<0.05).2.Hematoxylin-Eosin(HE)staining: pathological morphological changes in the CAl area of hippocampus in rats were observed under light microscope.In the sham operation group,the cells were distinct,no obvious damage was observed,the cell structure was intact,the nuclei were large and round,and the staining was uniform.In the model group,the cells in the hippocampal CAl area were disordered in arrangement,structural destruction,cell edema,vacuolar formation,cell shrinkage,nuclear condensation,chromatin edge collection,dark staining and uneven staining.Compared with the model group,the number of cell layers in the hippocampal CA1 area of the positive control group increased and the cells were closely arranged,and the pathological morphology was improved at different degree.Compared with the model group,the denaturation of neurons in the yishenjiainao group was less,the arrangement was relatively orderly,and the cell edema,vacuoles and nuclear shrinkage were significantly reduced.3.The expression of Nrf2(nucleus)and GPX4 protein in hippocampus:Western blot results showed that compared with the sham operation group,the expression of GPX4 protein in Nrf2(nucleus)of the model group decreased,with a significant difference(P <0.01)Compared with the model group,the positive control group Nrf2(nucleus),GPX4 protein expression increased,the difference was statistically significant(P <0.01);compared with the model group,the traditional Chinese medicine group Nrf2(nucleus),GPX4 protein expression The difference was statistically significant(P <0.05,P <0.01).Conclusion:1.Modified permanent bilateral common carotid artery ligation method to prepare VD rat model can better replicate the behavioral and pathological changes of VD rat.It is the currently recognized VD modeling method and the model is reliable.2.Yishenjiannao prescription and western medicine Donepezil can improve the learning and memory ability of VD rats,improve the pathological damage of nerve cells,up-regulate the expression of Nrf2 cell nucleus,and activate the downstream GPX4 protein expression.3.Yishenjiannao prescription may increase the scavenging of free radicals through the Nrf2 / GPX4 pathway,and improve the oxidative stress damage in the brain tissue of VD rats. |
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