Table 1.
The GEO dataset information.
Fig 1.
Normalized gene expression and principal component analysis.
(A) The normalization of GSE109227 dataset. (B) The principal component analysis of GSE109227. (C) The normalization of GSE65146 dataset. (D) The principal component analysis of GSE65146. (E) The normalization of GSE3644 dataset. (F) The Principal component analysis of GSE3644. AP, acute pancreatitis.
Fig 2.
Identification of differentially expressed genes (DEGs) and GO analysis.
Ranking plots for DEGs from three GEO datasets. Genes were sorted based on log2(FC) values, and DEGs with log2(FC) > 1 were selected. (A) Ranking of differentially expressed genes in GSE109227. (B) Ranking of differentially expressed genes in GSE65146. (C) Ranking of differentially expressed genes in GSE3644. The top 10 genes with the highest differential expression are marked in the plots. (D) Common DEGs identified by intersecting the three GEO datasets. (E) GO analysis of DEGs. GSEA analysis of ARG sets in the GEO dataset. (F) GSEA on ARG set in GSE3644. (G) GSEA on ARG set in GSE65146. (H) GSEA on ARG set in GSE109227. GSEA, gene set enrichment analysis; ARG, autophagy-related genes; Pval, P-value; NES, normalized enrichment score.
Fig 3.
Identification of differentially expressed autophagy-related genes (DEARGs) and functional analysis.
(A) DEARGs determined by intersecting DEGs with autophagy-related genes. (B) The volcano of DEARGs in GSE109227. (C) The volcano of DEARGs in GSE65146. (D) The volcano of DEARGs in GSE3644. (E) Gene-gene interaction network of DEARGs. Each node represents a gene, and the size of the node indicates the strength of the interaction. The color of the lines connecting nodes represents the type of gene interaction. The color of the nodes indicates the possible function of each gene. (F) GO enrichment analysis of DEARGs and 20 co-expressed genes. (G) KEGG enrichment analysis of DEARGs and 20 co-expressed genes. (H) Bubble diagram of GO enrichment term.
Fig 4.
Validation of the mRNA expression of 7 DEARGs in GSE121038.
The mRNAs of 6 DEARGs were significantly increased. (A) Sesn2, (B) Hmox1, (C) Nfe2l2F, (D) Cdkn1a, (E) Kras, (F) Cast. (G) Npc1 was not statistically significant compared to the sham group.
Fig 5.
Animal model of acute pancreatitis was successfully constructed.
(A) Pancreatic wet-to-dry weight ratio. (B) Serum amylase levels. (C) Representative images of pancreatic damage by H&E staining. Arrows indicate inflammatory cells, and asterisks indicate edematous stroma. (D) Western blot analysis was performed to assess the expression levels of DEARGs. (E) calpastatin, Nrf2, Npc1, (F) Sestrin2, p21, Kras, and (G) Ho1. Data shown are means ± SEM. NS: saline control, CAE: Caerulein, LPS: Lipopolysaccharides. *P < 0.05; **P < 0.01; ***P < 0.001; ns, no significant difference.
Fig 6.
Using the CTD database to construct a drug-gene interaction network with 7 DEGARs.
(A) An UpSetR plot shows that there are 2 drugs that regulate all genes. (B) Represents the relationship between existing drugs and the expression levels of DEARGs. Red and green arrows indicate that drugs will increase or decrease the expression of genes, respectively. Black arrows indicate that drugs will affect the expression of genes. Binding of potential drugs to DEARGs by molecular docking. (C) Binding mode of Acetaminophen to SESN2. (D) Binding mode of Acetaminophen to KRAS. (E) Binding mode of Acetaminophen to NPC1. (F) Binding mode of Acetaminophen to CAST. (G) Binding mode of Acetaminophen to HMOX1. (H)Binding mode of Acetaminophen to CDKN1A. (I) Binding mode of Acetaminophen toNFE2L2. (J) Heatmap of Docking Score. Compound is shown as cyan sticks. The key residues are shown as brick red sticks. Hydrogen bonds are shown as yellow dashed lines.
Fig 7.
Effects of Acetaminophen on DEARGs in AR42J Cells Treated with caerulein.
(A) AR42J cells were exposed to acetaminophen at concentrations ranging from 0 to 1800 µM for 24 hours. Cell viability was assessed using the CCK-8 assay, and the half-maximal inhibitory concentration (IC50) values were calculated. (B) Lipase activity was measured in different cell groups using a lipase assay kit. N = 3, *p < 0.05, **p < 0.01. (C) Western blot analysis was performed to assess the expression levels of calpastatin, Nrf2, Npc1, Sestrin2, p21, Kras, and Ho1, with β-actin as the loading control.