A Study On Molecular Mechanism Of Bilirubin-induced Neuronal Damage Based On A Transcriptome Analysis

Objective: Transcriptome sequencing technology was used to observe the differential expression of genes caused by unconjugated bilirubin in rat primary neuronal cell culture,revealing the mechanism of key biological processes or signaling pathways in the pathogenesis of bilirubin encephalopathy and discovering new mechanism of UCB-mediated neurotoxicity,providing a scientific basis for exploring new targets and interventions for the prevention and treatment of bilirubin encephalopathy.Methods: MTT detection of neuronal cell viability in bilirubin treated and control groups.Transcript sequencing analysis of primary cultured cortical neurons after bilirubin treatment.Results:(1)MTT test showed that UCB reduced cell viability of the neurons.With the increase of bilirubin concentration,the nerve cell viability decreased.(2)There were 140 differentially expressed genes between the experimental group and the control group,of which 117 were up-regulated genes and 23 were down-regulated genes.A total of 135 genes were annotated by database.In the COG classification,lipid transport and metabolism accounted for the largest proportion,ranging from 14 to 34.15%.In the KEGG annotation of differentially expressed genes,the number of differentially expressed genes in the metabolic pathway of steroid biosynthesis is the highest.Enrichment analysis of the KEGG pathway revealed that the differentially expressed genes enriched in the steroid biosynthetic pathway are the most significant.Cholesterol belongs to steroids,which has caused interest in studying cholesterol.To avoid omissions,after careful searching,other differentially expressed genes involved in cholesterol biosynthesis were also found.A total of 13 differentially expressed genes were involved in cholesterol biosynthesis and these genes were all up-regulated.Conclusion: After database annotation and enrichment analysis,13 genes involved in cholesterol biosynthesis were shown to be up-regulated,with a large number and a significant level of enrichment.Cholesterol synthesis can be used as a way to explore the mechanism of toxicity of bilirubin neurons,and the relevant genes are validated in subsequent studies.