Ya-Jing Guo, Jing Wen, Jing-Xia Liu, Yi-Ning Gao, Xin-Wang Chen, School of Acupuncture-Moxibustion and Tuina, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China
Ya-Jing Guo, The First Affiliated Hospital of Henan University of Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou 450014, Henan Province, China
Xin-Wang Chen, The Third Affiliated Hospital of Henan University of Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou 450008, Henan Province, China
Author contributions: Guo YJ and Chen XW jointly conceived and designed the study; Guo YJ drafted the manuscript; Wen J established the animal models and performed the related biological experiments; Liu JX conducted data analysis and prepared the figures; Gao YN verified the analytical results; Gao YN and Liu JX have seen and can confirm the authenticity of the raw data; all authors reviewed, edited, and approved the final manuscript.
AI contribution statement: Portions of this manuscript were edited using AI tools solely for checking grammar and typographical errors. The authors carefully reviewed and verified all AI-assisted outputs and take full responsibility for the scientific content of the manuscript.
Supported by National Natural Science Foundation of China, No. 82405585; Henan Province Science and Technology Research Project, No. 252102311264; Henan Higher Education Key Research Project, No. 23A360019; and China Postdoctoral Science Foundation, No. 2025M773941.
Institutional review board statement: This study does not involve any human experiments.
Institutional animal care and use committee statement: The animal experiment protocol of this study has been reviewed and approved by Laboratory Animal Welfare and Ethics Committee of Henan University of Traditional Chinese Medicine (now Henan University of Chinese Medicine) (approval No. IACUC-202310037).
Conflict-of-interest statement: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Corresponding author: Xin-Wang Chen, PhD, Associate Professor, School of Acupuncture-Moxibustion and Tuina, Henan University of Chinese Medicine, No. 156 Jinshui East Road, Zhengdong New District, Zhengzhou 450046, Henan Province, China. 787746907@qq.com
Received: May 25, 2026 Revised: July 24, 2026 Accepted: September 8, 2026 Published online: January 7, 2027 Processing time: 177 Days and 12.6 Hours
Abstract
BACKGROUND
Crohn’s disease (CD), a subtype of inflammatory bowel disease, is characterized by chronic inflammation affecting the entire gastrointestinal tract. Intestinal damage and subsequent disability caused by CD represent a growing global health concern, yet the molecular basis underlying its segmental heterogeneity remains poorly understood.
AIM
To construct a regional acetylome within different colonic segments and to determine the acetylation response to ileocolonic inflammation.
METHODS
Using colon tissue from distinct anatomical segments of CD rat models and normal controls, we performed quantitative acetyl-proteomics based on antibody enrichment coupled with mass spectrometry, as well as 16S sequencing of the gut microbiota. Targeted parallel reaction monitoring (PRM) was employed to validate the reliability of the discovery proteomics.
RESULTS
We constructed a spatial landscape of the colonic acetylome, identifying 4457 acetylation sites within 1476 acetylated proteins. Immune-regulatory signaling pathways were enriched, including Nucleotide-binding oligomerization domain-containing protein 2-receptor-interacting serine/threonine-protein kinase 2, mitogen-activated protein kinase, nuclear factor kappa-B and signal transducer and activator of transcription 3-Janus Kinase. Bioinformatic analyses integrating acetylome and microbiome data further revealed acetylation-associated alterations and their interactions with the gut microbiota, which may contribute to dysbiosis, providing new insights into CD’s segmental heterogeneity. PRM validation confirmed significant changes of selected acetylation sites including Ywhaz K3ac and SMRD2 K185ac.
CONCLUSION
Our findings suggest that multiple signaling cascades and acetylation events may contribute to disease progression, and these acetylation patterns may serve as a resource for future mechanistic and translational investigations.
Core Tip: This study presents a segment-resolved acetyl-proteomic landscape in a rat model of Crohn’s disease, revealing distinct acetylation signatures across the proximal, mid, and distal colon. Fifteen hub proteins, including receptor-interacting serine/threonine-protein kinase 2, signal transducer and activator of transcription 1, nuclear factor kappa B subunit 2, Ywhaz and SMRD2 were identified from the acetyl-protein interaction network. Site-specific validation by parallel reaction monitoring confirmed the reliability of the discovery proteomics. These region-specific acetylation patterns and hub proteins provide a foundation for biomarker discovery and targeted therapeutic strategies in Crohn’s disease.
