iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment

Yi Jiang,1,2,* Yingxue Bian,3,* Naqi Lian,1,2 Yaoqi Wang,1,2 Keliang Xie,1,2 Chao Qin,1,2 Yonghao Yu1,2 1Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin, People’s Republic of China; 2Tianjin Institute of Anesthesiology, Tianjin, People’s Republic of China; 3Department of Anesthesiology, Tianjin Union Medical Center, Tianjin, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yonghao Yu; Chao QinDepartment of Anesthesiology, Tianjin Medical University General Hospital, No. 154 Anshan Road, Heping District, Tianjin 300052, People’s Republic of ChinaEmail yuyonghao@126.com; qinchao0828@126.comObjective: Sepsis-associated intestinal injury has a higher morbidity and mortality in patients with sepsis, but there is still no effective treatment. Our research team has proven that inhaling 2% hydrogen gas (H2) can effectively improve sepsis and related organ damage, but the specific molecular mechanism of its role is not clear. In this study, isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics analysis was used for studying the effect of H2 on intestinal injury in sepsis.Methods: Male C57BL/6J mice were used to prepare a sepsis model by cecal ligation and puncture (CLP). The 7-day survival rates of mice were measured. 4-kd fluorescein isothiocyanate-conjugated Dextran (FITC-dextran) blood concentration measurement, combined with hematoxylin-eosinstain (HE) staining and Western blotting, was used to study the effect of H2 on sepsis-related intestinal damage. iTRAQ-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was used for studying the proteomics associated with H2 for the treatment of intestinal injury.Results: H2 can significantly improve the 7-day survival rates of sepsis mice. The load of blood and peritoneal lavage bacteria was increased, and H2 treatment can significantly reduce it. CLP mice had