Influence Of BXD Capsule On VEGF And Flk-1Expression In Hippocampus And Frontal Cortex Of Anger-out Model Rats

The relationship between emotions and disease, especially negative emotion isalready a focal point of nowadays society. Anger is one of the most serious one of theseven emotions of traditional Chinese medicine, which is also the most closely withemotion disease. There are two kinds of anger emotion according to the pattern ofmanifestation, and respectively named as “Anger-out” and “Anger-in”. Researchers ofwestern medicine know little about the central mechanism of “Anger-out” and “Anger-in”,and some researchers propose that “Anger-out” and “Anger-in” is important to thepathomechanism of anxiety and depression. All kinds of emotional diseases have effect oncentral nervous system and maybe lead to neuronal apoptosis. Recent years, VEGF receiveextensive attention in the research of neurotrophy and neuroprotection, and its researchesin kinds of diseases, such as cerebral ischemia, tumour, alzheimer disease have alreadymade some progress. There are also some progress in the study of depression and anxietyabroad, which is deficient inland.ObjectiveTo test and contrast Hippocampus and Frontal VEGF content change and Flk-1expression change in the onset and progress of “Anger-out” emotion on the carrier of therat model and drug intervention, so as to explore the mechanisms of VEGF signalingpathways in the onset and progress of “Anger-out” emotion, as well as provide datasupport for the complete of medicinal mechanism of liver medicine.Methods1Model prepare and evaluation96rats with similar open-field test score were selected form120SPF male wistar rats, 12of which were selected as Control group and breed normally. The half of last84wereprepared for “Anger-out” model and divided into3groups randomly: model group, prozacgroup and BXD groups (n1=n2=n3=14). Macroscopic behavior evaluation between groupswas compared before and after drugs.2Tissue extractionsAll rats were beheaded in the last day of model prepare, which hippocampus andfrontal cortex were extracted and stored in-70℃ultra-low temperature freezer waitingfor inspection.3microscopic index testHippocampus and frontal cortex VEGF contents of the rats were detected with ELISAkit; at the same time, immunofluorescence was used to observe the distribution of Flk-1inhippocampus and frontal tissue. Further more, mRNA expression of VEGF and Flk-1wasdetermined with conventional RT-PCR method; and protein expression of Flk-1wasdetermined with conventional Western blot method.Results1macroscopic behavior evaluation result⑴Weight: Before drug, there were no significant differences in the initial weight and theadaptive weight between groups. After2weeks’ of drug, Model rats were significantlyreduced compared to Control group (P<0.05), but not BXD group and Prozac group.⑵Open-field test score: Before modeling, there were no significant differences inOpen-field test score between groups. After1weeks’ of drug, Model rats weresignificantly increased (P<0.05), but not BXD group and Prozac group.⑶Sugar water preference coefficient：Before modeling, there were no significantdifferences in sugar water preference coefficient between groups. After1weeks’ of drug,Model rats were significantly reduced (P<0.05), but not BXD group and Prozac group.⑷Aggressive behaviour score: Before modeling, there were no significant differences inaggressive behaviour score between groups. After1weeks’ of drug, BXD group andProzac group were all reduced significantly (P<0.05).⑸Incubation period: Before modeling, there were no significant differences in incubation period between groups. After1weeks’ of drug, BXD group and Prozac groupwere all increased significantly (P<0.05).2microscopic index measurement results(1) Various brain regions’Flk-1distribution and morphological changes: Bothhippocampus and frontal area have rich distribution of Flk-1. The normal pyramidal cellsof CA3were closely packed neatly; while the pyramidal cells of CA3in model group havebigger gap, and some cells arranged crooked; which improved after drugs. Frontal areacells of normal rats distributed homogeneously, and for dot or cone; while the frontal areacells of model rats seemed thin and elliptic; which improved after drugs too.(2) Flk-1mRNA expression: Compared to Control group, model group was significantlylower in Flk-1mRNA expression (P<0.05); Compared to model group, both BXD groupand Prozac group were significantly higher in Flk-1mRNA expression (P<0.05).(3) Flk-1protein expression: Compared to Control group, model group was significantlylower in Flk-1protein expression (P<0.05); Compared to modal group, both BXD groupand Prozac group were significantly higher in Flk-1protein expression (P<0.05).(4) VEGF contents: Compared to Control group, Model group was significantly lower inVEGF contents (P<0.05); Compared to Model group, Prozac group and BXD group wassignificantly was significantly higher in VEGF contents (P <0.05).Conclusions1The model prepare method of social isolation and invasion for “Anger-out” rats isfeasible.2Both hippocampus and frontal area have rich distribution of Flk-1.3The “Anger-out” model rats has pathological characteristics of deficiency VEGF innervous centralis and down regulated Flk-1expression in central, which may be animportant reason for neuronal apoptosis and nerve injury, and closely related with itspathogenesis.4Both BXD and fluoxetine have positive effect on the macroscopic behaviors of “Fennu”modal rats, which also have ability to increase VEGF and up regulate Flk-1expression. The VEGF signal path may be one of the important targets for the therapy effect of liverdrugs.