SCREENING FOR KEY GENES INVOLVED IN PROPOFOL-INDUCED NEUROTOXICITY BASED ON METABOLOMICS AND TRANSCRIPTOMICS

Abstract

Objective To screen for the key genes involved in propofol-induced neurotoxicity based on metabolomics and transcriptomics. Methods HT22 cells were cultured with propofol at concentrations of 0, 25, 50, 75, and 100 mg/L for 24 h and were established as Ctrl, P25, P50, P75, and P100 groups, respectively, and CCK-9 assay was used to measure cell viability in each group. HT22 cells were collected after 24 h of treatment to extract RNA and metabolites for metabolite detection and transcriptome analysis. The extracted HT22 cell metabolites were analyzed by liquid chromatography-mass spectrometry (LC-MS), differentially expressed metabolites, and trend analysis to obtain the key metabolites. The RNA of HT22 cells was analyzed by sequence alignment and differentially expressed genes to obtain the differentially expressed genes. Weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) analysis were performed for the above key metabolites and differentially expressed genes to obtain the key genes involved in propofol-induced neurotoxicity. Results The viability of HT22 cells decreased with the increase in propofol concentration (t=11.40-97.25,P<0.05). LC-MS was performed for the metabolites of HT22 cells and a total of 2 701 metabolites were identified, and 49 key metabolites were obtained through further differential ana-lysis and trend analysis. A sequence alignment analysis of the extracted HT22 cell RNA showed that the number of sequences in each group of HT22 cells could meet the requirements for data analysis, and there were 1 254 differentially expressed genes between the Ctrl group and the P25 group, 4 695 differentially expressed genes between the Ctrl group and the P50 group, 4 923 differentially expressed genes between the Ctrl group and the P75 group, and 5 031 differentially expressed genes between the Ctrl group and the P100 group. WGCNA and PPI analyses were performed for the key metabolites and the differentially expressed genes, and a total of 15 key genes were obtained, namely NTNG2, ENG, SEMA4G, JAG2, FGF11, SERPINE1, GDF15, GADD45G, F3, NGF, FGF21, PGF, EGFL7, SEMA6D, and LRFN1. Conclusion The key genes involved in propofol-induced neurotoxicity may include NTNG2, ENG, SEMA4G, JAG2, FGF11, SERPINE1, GDF15, GADD45G, F3, NGF, FGF21, PGF, EGFL7, SEMA6D, and LRFN1. These genes provide a theoretical basis for the subsequent research on the molecular mechanism of propofol-induced neurotoxicity and research directions for the development of preventive and therapeutic drugs for propofol-induced neurotoxicity.

Keywords

• hippocampus|neurons|propofol|nervous system|toxic actions|metabolomics|transcriptome