Data-Independent Acquisition-Based Mass Spectrometry (DIA-MS) for Quantitative Analysis of Human Intestinal Ischemia/Reperfusion

Intestinal ischemia–reperfusion (II/R) injury is a complex pathologic process, which is of great significance to unravel the underlying mechanisms and pathophysiology. Our study represented a comprehensive proteomic analysis in the human intestine with ischemia–reperfusion injury. The proteomics analysis measured a total of 5,230 proteins, and 417 differently expressed proteins (DEPs) were identified between II/R and control samples. GO and KEGG analysis demonstrated that the 290 upregulated DEPs in II/R were significantly involved in immune-related biological process and tight junction, focal adhesion, and cAMP signaling pathway, whereas the 127 downregulated DEPs in II/R were enriched in lipid metabolic process and metabolic pathway. Furthermore, we screened out 20 hub proteins from the protein–protein interaction (PPI) network according to the degree of connectivity, and six clusters were identified. Combined with the result of KEGG analysis, 6 from the 20 hub proteins, ACTB, CAV1, FLNA, MYLK, ACTN1, and MYL9, were identified as the key proteins in the progress of II/R injury. According to the previous studies, FLNA and MYL9 were selected as the novel disease-related proteins for the first time. In conclusion, this study extended our understanding of the alteration in the human intestine during ischemia and reperfusion and highlighted the potential role of FLNA and MYL9 in the progress of II/R injury, which need to be further studied.