Canine Small Intestinal Epithelial Cells

Analysis of Pathway Enrichment of the Response of Canine Intestinal Epithelial Cells Treated by Sulforaphane

Sulforaphane (SFN), a compound from cruciferous vegetables, is known for its anti-inflammatory and antioxidant properties, particularly beneficial for intestinal health. It can modulate inflammatory responses and enhance intestinal cell regeneration. Wang's team explored SFN's effects on canine small intestinal epithelial cells, focusing on its molecular impact through transcriptome analysis to understand its role in modulating biological processes involved in inflammation and oxidative stress.

To pinpoint SFN's regulatory transcriptional pathways, they conducted a transcriptome analysis on canine small intestinal epithelial cells, using SFN (4 μM) and Vehicle groups. They filtered the fold change of FPKM for valid data and performed enrichment analysis with GSEA 4.1.0. Key enrichments in the SFN group included the Inflammatory response, Lipid biosynthesis, Oxidative stress response, and T-cell immunity pathways (Fig. 1A), with the "Inflammatory response" pathway having the highest significance. Up-regulated genes focused on Protein digestion, PPAR signaling, and Fatty acid biosynthesis in KEGG (Fig. 1B and C). Down-regulated genes involved Cytokine interaction, MAPK signaling, and IL-17 pathways (Fig. 1D and E). Up-regulated DEGs also enriched in tissue and protein structure pathways, whereas down-regulated ones related to cytoarchitecture (Fig. 1F).

Fig. 1. GSEA, KEGG and GO enrichment analysis of DEGs. Showing GSEA enrichment plot, enrichment bubble plot, Cnetplot and GO enrichment circle plot (Li K, Yan J, et al., 2024).

MLT and Cell Growth, Survival, and KEGG Enrichment in cIECs

Melatonin (MLT), a hormone produced by the pineal gland, plays a crucial role in regulating inflammation in mammals. Hong's team investigates how melatonin influences gene expression in canine ileum epithelial cells (Canine small intestinal epithelial cells, cIECs), focusing on the modulation of inflammatory genes ZAP70 and CD40 through histone modifications.

To evaluate the protective effects of a 10 μM MLT extract on cIECs, cell viability and apoptosis were assessed. No significant differences in cell numbers or caspase 3/7 activity, an apoptosis indicator, were observed between MLT-treated and control groups (Fig. 2A and B). Transcriptome analysis highlighted genes significantly affected by MLT (Fig. 2C and D), with GSEA identifying primary immunodeficiency and NF-kappa B signaling as key enriched pathways (Fig. 2E). No major changes were noted in cell cycle or proliferation genes (Fig. 2F). Functional annotation and pathway enrichment analysis (Fig. 3A-C) supported these findings, while the PPI network (Fig. 3D and E) showed the complex interactions and regulatory effects influenced by MLT on cIECs.

Figure 2. MLT treatment does not affect cell survival and growth in canine epithelial cells (Hong J, Adam SY, et al., 2025).

Fig. 3. Functional annotation pathway enrichment result (Hong J, Adam SY, et al., 2025).