ORIGINAL RESEARCH article
Front. Immunol.
Sec. Inflammation
Bioinformatics-Driven Insights: Rapamycin-Mediated CaMK2D Inhibition Alleviates Intestinal Ischemia-Reperfusion Injury
Provisionally accepted
- The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, China
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Introduction: Intestinal ischemia-reperfusion (I/R) injury, a common and severe clinical condition with high morbidity and mortality, burdens healthcare systems. Our previous investigations established that a nano-delivery system enabled targeted rapamycin delivery to intestinal I/R injury sites with therapeutic efficacy. While calcium/calmodulin-dependent protein kinase IIδ (CaMK2D) has been implicated in myocardial injury and tumorigenesis, its role in intestinal I/R pathophysiology remains unexplored. This study investigates the therapeutic mechanisms of rapamycin in intestinal I/R injury by modulation of CaMK2D signaling. Methods: An oxygen-glucose deprivation/reperfusion (OGD/R) model in Caco-2 human colorectal cancer cells and a murine intestinal I/R model were established. Small interfering RNA (siRNA) and hesperadin (HES) were used to inhibit CaMK2D expression. Transcriptomic profiling was performed via RNA sequencing (RNA-Seq) with subsequent bioinformatic analysis including differential gene expression, MCODE-based protein interaction network clustering, and RAPA-CaMK2D molecular docking studies. Cellular assays included qRT - PCR, western blotting (WB), Fluo-3 calcium flux analysis, flow cytometry, and Enzyme-linked immunosorbent assay (ELISA). In animal experiments, HE staining, immunohistochemistry, TUNEL assay, WB, and ELISA were employed. Results: Both cellular and murine models demonstrated a significant upregulation of CaMK2D phosphorylation with intestinal epithelial apoptosis, barrier dysfunction, and enhanced inflammatory response during I/R. CaMK2D knockdown using siRNA attenuated these pathological manifestations, vice versa. Bioinformatic analysis revealed a CaMK2D-dominated regulatory module (ranked fifth) enriched in calcium-mediated sinaling pathways. Mechanistically, I/R induced CaMK2D activation exacerbated inflammatory cascades, epithelial apoptosis, and tight junction disruption. Rapamycin treatment (1.5 mg/kg, i.p.) ameliorated these effects by decreasing CaMK2D expression and phosphorylation (WB, P < 0.01), pro-inflammatory cytokine levels (ELISA, P < 0.01), while preserving intestinal integrity as evidenced by histological analysis (IHC, P < 0.05). Discussion: Our findings establish CaMK2D hyperactivation as a key to intestinal I/R injury. The therapeutic potential of rapamycin derived from its ability to suppress CaMK2D signaling axis, providing a novel pharmacological strategy for intestinal I/R management.
Keywords: Calcium/calmodulin dependent-protein kinase IIδ, Genomics, Inflammation, Intestinal ischemia-reperfusion injury, rapamycin
Received: 13 Aug 2025; Accepted: 26 Dec 2025.
Copyright: © 2025 Sheng, Liang, Zhang, Lai, Hong, Huang and Liu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Dezhao Liu
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