| Stress brain injury is an organic damage to the brain tissue caused by long-term psychological and environmental stress.Neuronal damage is one of the foundations for the formation of neurological diseases such as depression and Parkinson’s disease.However,the mechanism of stress-induced brain injury is not clear.Here,we use transcriptome sequencing(RNA-seq),bioinformatics analysis tools such as GO and KEGG to discover and screen differentially expressed genes.Significantly differentially expressed genes and their enriched signaling pathways were systematically studied.The molecular mechanism leading to stress brain injury was summarized,and the key molecules differentially expressed were verified via experiments.We aim to provide experimental basis for prevention and treatment of stress brain injury and discovery of new drug treatment targets.Part 1: Establishment of a rat model of brain injury indeced by chronic stressA 21-day model of stress-induced brain injury was established in rats by water immersion restraint,and the model was verified by body weight,Nissl staining,serological indicators,behavioral experiments and Immunohistochemistry.The results showed that after 21 days of chronic stress,the body weight and growth rate of the rats in the model group decreased significantly.In addition,serum corticosterone and norepinephrine levels increased.The open field test showed that the total movement distance,average speed and raering times of the rats in the model group decreased,and the immobility time increased.Forced swimming test also showed an increase immobility time.Nissl staining showed that Nissl bodies in the CA1 and CA3 area of the brain tissue of the model group were reduced and the nuclei was lost.Immunohistochemistry showed that,the expression of c-Fos in hippocampus of model group was increased significantly.The results suggested that chronic stress leads to behavioral disorders and brain injury in the rats of model group.The chronic stress brain injury model was successfully established.Part 2: Transcriptome Analysis of Brain Tissue in Stress Brain Injury RatsThe m RNA in the control group and the model group was extracted,purified,identified for integrity,constructed and sequenced to obtain differential gene expression profiles.At the same time,q RT-PCR was used to verify differentially expressed genes.GO and KEGG were used to analyze the signal pathways related to the enrichment of differentially expressed genes.The results showed that in stress-induced brain injury tissues,the expression of voltage-gated potassium channel-related genes was significantly increased.Voltage-gated potassium channels are potential therapeutic targets for stress-induced brain injury,and voltage-gated potassium channel openers have potential anti-stress protection.Part 3: The anti-stress protective effect of K channel opener RetigabineThrough the analysis of transcriptome sequencing data,K channel opener Retigabine was selected for interventional treatment of rats with stress brain injury.Retigabine’s therapeutic effect was verified through behavioral experiments,serological indicators,Nissl staining,immunohistochemistry and western blotting.The results showed that after the treatment with Retigabine at a dose of 4 mg / kg,the body weight and growth rate of the rats increased significantly.Besides,serum corticosterone and norepinephrine levels decreased significantly.The open field test showed that compared with the model group,the total movement distance,average speed,and rearing times of the rats increased,and the immobility time decreased after the treatment of Retigabine at a dose of 4 mg / kg.Forced swimming test showed that the immobility time also reduced.Nissl staining showed that after treatment with Retigabine at a dose of 4 mg / kg,Nissl bodies in rat brain tissue increased.Immunohistochemical results showed that c-Fos was significantly reduced under Retigabine treatment at a dose of 4 mg / kg.It shows that Retigabine has significant anti-stress protection effect.Western Blotting showed that at 4 mg / kg Retigabine,the expression of apoptosis-related proteins Caspase3,cleaved-caspase3 and Bax were decreased,and Bcl-2expression increased,suggesting that Retigabine plays an anti-stress protective role via reducing neuronal apoptosis.In summary,the transcriptome analysis of the brain tissue of a model of stress-induced brain injury showed that the blockage of potassium channels was one of the reasons of stress-induced brain injury.Therefore,potassium channels may be an important drug target for the development of anti-stress drugs,and potassium channel openers will become candidates for anti-SBI. |
PDF Full Text Request