The Impact of E. coli HPI on the PI3K/Akt/mTOR Autophagy Pathway in Mouse M Cells, Intestinal Tissue, and Macrophages

This dataset is derived from a study designed to explore the pathogenic mechanism of the high-pathogenicity island (HPI) of pathogenic Escherichia coli (E. coli) and its interaction with host cell autophagy. The core research hypothesis is that the HPI, a major virulence factor of pathogenic E. coli, modulates the autophagic response of host cells by regulating the PI3K/Akt/mTOR signaling pathway, thereby further altering the inflammatory microenvironment associated with E. coli infection and the degree of host cellular and tissue injury. Specifically, we hypothesized that HPI could either promote or inhibit autophagy through the PI3K/Akt/mTOR pathway, and this regulation of autophagy would play a key role in mediating the pathological effects of HPI on host intestinal tissue, M cells, and macrophages.The dataset comprehensively includes multi-dimensional experimental data from both in vitro cellular experiments and in vivo animal models, covering the key indicators of HPI-induced autophagy regulation, PI3K/Akt/mTOR pathway activity, inflammatory response, and tissue injury. The specific content and corresponding experimental contexts are detailed as follows: Autophagic flux-related data: Protein expression levels of autophagy markers (LC3-I, LC3-II, Beclin-1, and p62) detected by Western blotting; relative mRNA expression levels of autophagy-related genes (if detected) measured by qPCR; and ultra-structural images of autophagosomes in RAW264.7 cells observed by electron microscopy, which are used to quantify the number and morphology of autophagosomes. PI3K/Akt/mTOR pathway activity data: Protein expression levels of key pathway molecules (PI3K, Akt, mTOR) and their phosphorylated forms (p-PI3K, p-Akt, p-mTOR) detected by Western blotting, reflecting the activation status of the pathway. Infection and intervention data: Bacterial load in RAW264.7 cells after infection with wild-type HPI-positive (HPI+) E. coli and HPI-deficient mutant (Δirp2/ΔHPI) E. coli; changes in autophagy and pathway activity after treatment with autophagy modulators (Rapamycin as an autophagy activator, 3-MA as an autophagy inhibitor) and Beclin-1-silenced RAW264.7 cells; relative mRNA expression levels of pro-inflammatory cytokines (IL-1β, TNF-α) in infected cells detected by qPCR.- Intestinal tissue-related data: Expression levels of intestinal M-cell markers (detected by qPCR or immunohistochemistry) and secretory immunoglobulin A (sIgA) levels in intestinal mucosa (detected by enzyme-linked immunosorbent assay, ELISA) after infection with HPI+ or ΔHPI E. coli. Inflammatory response data: Concentrations of pro-inflammatory cytokines (IL-1β, TNF-α, IFN-γ) in mouse serum or intestinal tissue homogenates detected by ELISA, reflecting the systemic and local inflammatory status induced by HPI. Tissue pathology data: