Sevoflurane upregulates neuron death process-related Ddit4 expression by NMDAR in the hippocampus

Figures

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  Figure 1. Differentially regulated genes between the control and sevoflurane-treated groups. (A) Boxplot of all genes in each sample. (B) Heatmap of differentially regulated genes in the control and sevoflurane-treated groups, 194 upregulated and 32 downregulated genes after treatment with 2.5% sevoflurane in 100% oxygen for 4 hours. (Green: Control group; Red: sevoflurane-treated group).

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  Figure 2. GO and KEGG enrichment analyses of differentially regulated genes. (A) Functions of biological processes that are significantly enriched by differentially expressed genes. (B) Top 20 significantly enriched biological processes. (C) Ten KEGG pathways were significantly enriched by differentially expressed genes.

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  Figure 3. Cell death-related biological processes were significantly enriched by differentially expressed genes. (A) Five cell death-related biological processes were significantly enriched by differentially expressed genes. (B) Thirteen differentially expressed genes (10 genes upregulated in the sevoflurane-treated group and 3 genes downregulated in the sevoflurane-treated group) were significantly enriched in positive regulation of cell death. (C) Ten differentially expressed genes (8 genes upregulated in the sevoflurane-treated group and 2 genes downregulated in the sevoflurane-treated group) were significantly enriched in the regulation of neuronal death. (D) Seven differentially expressed genes (6 genes upregulated in the sevoflurane-treated group and 1 gene downregulated in the sevoflurane-treated group) were significantly enriched in the negative regulation of neuronal death. (E) Nine differentially expressed genes (7 genes upregulated in the sevoflurane-treated group and 2 genes downregulated in the sevoflurane-treated group) were significantly enriched in the regulation of neuronal apoptosis. (F) Seven differentially expressed genes (6 genes upregulated in the sevoflurane-treated…

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  Figure 4. PPI network of differentially regulated genes in STRING. (Red: upregulated differentially expressed genes in the sevoflurane-treated group; green: downregulated differentially expressed genes in the sevoflurane-treated group).

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  Figure 5. Six hub genes were enriched in biological process terms, and 4 hub genes were associated with cell death biological process terms. (A) Six hub genes enriched in biological process terms. (B) Expression of 6 hub genes between the control and sevoflurane-treated groups. (C) qPCR detection of Agt, Cdkn1a, Ddit4, Pdgfra, Rapgef3, and Rhob mRNA expression levels in the hippocampus of the mice that received 3% sevoflurane exposure for 4 h or the control mice. Sevoflurane indicates the mice received sevoflurane exposure. Control indicates that the mice were raised only under normal conditions. (D) Four hub genes associated with cell death biological process terms. The data shown are the means ± SDs, n = 3. *P < 0.05, **P < 0.01. (Green: Control group; Red: sevoflurane-treated group).

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  Figure 6. Hub genes associated with cell death biological process terms. (A) Four hub genes enriched in cell death biological process terms. (B) qPCR detection of the Ddit4 mRNA level in the hippocampus of mice with hippocampal stereotactic injection of muscimol or saline. (C) qPCR detection of the Ddit4 mRNA level in the hippocampus of mice with the hippocampal stereotactic injection of MK-801 or saline. (D) qPCR detection indicated the Ddit4 mRNA level in the hippocampus of mice that received sevoflurane exposure with NMDA (sevoflurane + NMDA group) or saline (sevoflurane group) injection into the hippocampus. Control indicates mice injected with saline in the hippocampus. MK-801 indicates mice injected with MK-801 in the hippocampus. (E) Representative pictures showed that DDIT4 level was elevated as shown by western blotting. The data shown are the means ± SDs, n = 3. nsP > 0.05, *P < 0.05.