| Objective: Fear memory induced by traumatic stress and its accompanying anxiety will lead to the decline of individual quality of life,impaired social functions,and often accompanied by drug abuse,phobia and nssi,etc.,which will cause long-term and lasting negative effects on family and society.Nucleus accumbens,as the information integration nucleus of basal ganglia,plays a pivotal role in regulating emotions and their related behaviors.The nucleus accumbens,as part of the parasymptomatic limbic system,is involved in forming the reward circuit and is involved in drug addiction.γ-aminobutyric acid neurons in nucleus accumbens and various neurotransmitters they receive are involved in learning,memory and emotional activity feedback regulation,which are converted into behavioral responses.At present,the molecular mechanism by which the nucleus accumbens regulates fear memory and anxiety remains unclear.Therefore,in-depth exploration of the molecular mechanism of stress and the search for effective therapeutic targets in the nucleus accumbens is of great guiding significance for the development of lasting and effective preventive and therapeutic measures.Methods: Male C57BL/6J mice were taken as the main research object,and the experimental mice were screened with avoidance test at the initial stage.The mice that met the requirements were divided into control group,observation group and invasion group,and were respectively treated with physical trauma stress and psychological stress.After the modeling,avoidance test was conducted.Compared with the control group,the nucleus accumbens of the mice with obvious fear memory and anxiety in the experimental group were measured with high throughput to screen out the differentially expressed genes.The sequencing results were analyzed by bioinformatics,and some genes were verified by q PCR and dual luciferase assay system.Results:The results were as follows: 1)after stress building five days later,after the invasion of group building relative to avoid before building the result value decreased significantly,observation group relative to the building after building before to circumvent the result value decreased significantly,the invasion of group compared to control group to evade the result value decreased significantly,the invasion of group compared with the observation way to circumvent the result value decreased significantly(P values are less than 0.05).2)by comparing the results of high-throughput sequencing of nucleus accumvae of mice in the control group,the invasion group and the viewing group,micro RNA and m RNA with differential expressions greater than 1.5 were screened out as the standard.Through invasion,watch group and control group of mice respectively,the nucleus accumbens compared brain regions of differentially expressed genes,encoding Adr R alpha,BDNF,SSR,ACh RM2/3,Glu RM2/8,the genes increase Hr R1 and AC,coding DR3/5,PR2,GP gamma 8 and P450 genes,these changes in synaptic receptors and signaling molecules that physical/mental stress or psychological stress results in the formation of a state of fear and anxiety.3)through the differentially expressed genes signaling pathway enrichment analysis showed neuronal activity ligand receptor interaction,calcium signaling pathways,WNT signaling pathways,dopaminergic synapses,neurotrophic factor signaling pathways,axon guidance,MAPK signal,can synaptic glutamate,cholinergic synaptic signaling pathway involved in physiological or psychological stress stress caused by the formation of fear memory.4)three target gene prediction data(Targetscan,RNAhybrid and mi Randa)were used to predict the target genes of differentially expressed micro RNA,which were further overlapped with the differentially expressed genes already constructed,and the regulatory network diagram of micro RNA/m RNA in nucleus accumaccumae of stressed mice was drawn.5)a part of differentially expressed micro RNA and m RNA were selected for q PCR verification of sequencing analysis results,further proving the accuracy of sequencing results(P values were less than 0.05).6)dual luciferase assay system showed that mi R-483-5p was directly targeted at Crhr1,mi R-669p-5p was directly targeted at Adralb,Crhr1 was negatively regulated by mi R-483-5p,and Adralb was negatively regulated by mi R-669p-5p.Conclusion: Stress response was produced by stress modeling in mice.Psychological/physiological stress or psychological stress in the nucleus accumbens through different synapses,signaling pathways and other ways led to the formation of fear memory and anxiety state;The sequencing results of m RNA and micro RNA in nucleus accumbialis were analyzed,and the consistency of sequencing results and analysis conclusions was verified by q PCR and dual luciferase assay system. |
