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 Republi...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Dove Medical Press
2020-11-01
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| Series: | Drug Design, Development and Therapy |
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| Online Access: | https://www.dovepress.com/itraq-based-quantitative-proteomic-analysis-of-intestines-in-murine-po-peer-reviewed-article-DDDT |
| _version_ | 1818655700934459392 |
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| author | Jiang Y Bian Y Lian N Wang Y Xie K Qin C Yu Y |
| author_facet | Jiang Y Bian Y Lian N Wang Y Xie K Qin C Yu Y |
| author_sort | Jiang Y |
| collection | DOAJ |
| description | 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 significant intestinal damage, and inhalation of 2% hydrogen could significantly reduce this damage. All 4194 proteins were quantified, of which 199 differentially expressed proteins were associated with the positive effect of H2 on sepsis. Functional enrichment analysis indicated that H2 may reduce intestinal injury in septic mice through the effects of thyroid hormone synthesis and nitrogen metabolism signaling pathway. Western blot showed that H2 was reduced by down-regulating the expressions of deleted in malignant brain tumors 1 protein (DMBT1), insulin receptor substrate 2 (IRS2), N-myc downregulated gene 1 (NDRG1) and serum amyloid A-1 protein (SAA1) intestinal damage in sepsis mice.Conclusion: A total of 199 differential proteins were related with H2 in the intestinal protection of sepsis. H2-related differential proteins were notably enriched in the following signaling pathways, including thyroid hormone synthesis signaling pathway, nitrogen metabolism signaling pathways, digestion and absorption signaling pathways (vitamins, proteins and fats). H2 reduced intestinal injury in septic mice by down-regulating the expressions of SAA1, NDRG1, DMBT1 and IRS2.Keywords: sepsis, intestinal injury, hydrogen gas, proteomics, isobaric tags for relative and absolute quantitation, iTRAQ |
| first_indexed | 2024-12-17T03:13:52Z |
| format | Article |
| id | doaj.art-b8ba2dd4ae7f408fbba3ce53b9aab284 |
| institution | Directory Open Access Journal |
| issn | 1177-8881 |
| language | English |
| last_indexed | 2024-12-17T03:13:52Z |
| publishDate | 2020-11-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | Drug Design, Development and Therapy |
| spelling | doaj.art-b8ba2dd4ae7f408fbba3ce53b9aab2842022-12-21T22:05:45ZengDove Medical PressDrug Design, Development and Therapy1177-88812020-11-01Volume 144885490059202iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas TreatmentJiang YBian YLian NWang YXie KQin CYu YYi 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 significant intestinal damage, and inhalation of 2% hydrogen could significantly reduce this damage. All 4194 proteins were quantified, of which 199 differentially expressed proteins were associated with the positive effect of H2 on sepsis. Functional enrichment analysis indicated that H2 may reduce intestinal injury in septic mice through the effects of thyroid hormone synthesis and nitrogen metabolism signaling pathway. Western blot showed that H2 was reduced by down-regulating the expressions of deleted in malignant brain tumors 1 protein (DMBT1), insulin receptor substrate 2 (IRS2), N-myc downregulated gene 1 (NDRG1) and serum amyloid A-1 protein (SAA1) intestinal damage in sepsis mice.Conclusion: A total of 199 differential proteins were related with H2 in the intestinal protection of sepsis. H2-related differential proteins were notably enriched in the following signaling pathways, including thyroid hormone synthesis signaling pathway, nitrogen metabolism signaling pathways, digestion and absorption signaling pathways (vitamins, proteins and fats). H2 reduced intestinal injury in septic mice by down-regulating the expressions of SAA1, NDRG1, DMBT1 and IRS2.Keywords: sepsis, intestinal injury, hydrogen gas, proteomics, isobaric tags for relative and absolute quantitation, iTRAQhttps://www.dovepress.com/itraq-based-quantitative-proteomic-analysis-of-intestines-in-murine-po-peer-reviewed-article-DDDTsepsisintestinal injuryhydrogen gasproteomicsisobaric tags for relative and absolute quantitation (itraq) |
| spellingShingle | Jiang Y Bian Y Lian N Wang Y Xie K Qin C Yu Y iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment Drug Design, Development and Therapy sepsis intestinal injury hydrogen gas proteomics isobaric tags for relative and absolute quantitation (itraq) |
| title | iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment |
| title_full | iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment |
| title_fullStr | iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment |
| title_full_unstemmed | iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment |
| title_short | iTRAQ-Based Quantitative Proteomic Analysis of Intestines in Murine Polymicrobial Sepsis with Hydrogen Gas Treatment |
| title_sort | itraq based quantitative proteomic analysis of intestines in murine polymicrobial sepsis with hydrogen gas treatment |
| topic | sepsis intestinal injury hydrogen gas proteomics isobaric tags for relative and absolute quantitation (itraq) |
| url | https://www.dovepress.com/itraq-based-quantitative-proteomic-analysis-of-intestines-in-murine-po-peer-reviewed-article-DDDT |
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