Transcriptome Analysis Revealed That Ischemic Post-conditioning Suppressed the Expression of Inflammatory Genes in Lung Ischemia-reperfusion Injury

Overview

Journal Front Genet

Date 2024 Dec 10

PMID 39655220

Authors

Liangen Lin

Congcong Sun

Yuanwen Ye

Peng Zhu

Keyue Pan

Linglong Chen

Affiliations

Soon will be listed here.

Abstract

Introduction: Ischemic post-conditioning (I-post C) is a recognized therapeutic strategy for lung ischemia/reperfusion injury (LIRI). However, the specific mechanisms underlying the lung protection conferred by I-post C remain unclear. This study aimed to investigate the protective mechanisms and potential molecular regulatory networks of I-post C on lung tissue.

Methods: Transcriptome analysis was performed on rat lung tissues obtained from Sham, ischemia-reperfusion (IR), and I-post C groups using RNA-seq to identify differentially expressed genes (DEGs). Subsequently, gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and gene set enrichment analysis (GSEA) were conducted to elucidate significantly enriched pathways in the IR and I-post C groups. Additionally, protein-protein interaction (PPI) network analysis was carried out to examine associations among the DEGs. Pathological changes in lung tissues were assessed using hematoxylin-eosin (H&E) staining. The expression levels of CXCL1 and CXCL6 in the IR and I-post C groups were evaluated through immunofluorescence and Western blotting.

Results: Our results showed that I-post C significantly attenuated both pulmonary edema and inflammatory cell infiltration. Transcriptome analysis identified 38 DEGs in the I-post C group compared to the IR group, comprising 21 upregulated and 17 downregulated genes. Among these, seven inflammation-related DEGs exhibited co-expression patterns with the Sham and IR groups, with notable downregulation of C and C. GO analysis primarily linked these DEGs to neutrophil activation, chemotaxis, cytokine activity, and CCR chemokine receptor binding. KEGG analysis revealed enriched pathways, including the IL-17, TNF, and NF-κB signaling pathways. GSEA indicated downregulation of neutrophil chemotaxis and the IL-17 signaling pathway, correlating with reduced expression of C and C. Validation of and mRNA expression via immunofluorescence and Western blotting supported the RNA-seq findings. Furthermore, a PPI network was constructed to elucidate interactions among the 29 DEGs.

Conclusions: Through RNA-Seq analysis, we concluded that I-post C may reduce inflammation and suppress the IL-17 signaling pathway, thereby protecting against lung damage caused by LIRI, potentially involving neutrophil extracellular traps.

References

Wang Y, Dong Z, Zhang Z, Wang Y, Yang K, Li X . Postconditioning with Irisin Attenuates Lung Ischemia/Reperfusion Injury by Suppressing Ferroptosis via Induction of the Nrf2/HO-1 Signal Axis. Oxid Med Cell Longev. 2022; 2022:9911167. PMC: 8906956. DOI: 10.1155/2022/9911167. View

De Filippo K, Dudeck A, Hasenberg M, Nye E, van Rooijen N, Hartmann K . Mast cell and macrophage chemokines CXCL1/CXCL2 control the early stage of neutrophil recruitment during tissue inflammation. Blood. 2013; 121(24):4930-7. DOI: 10.1182/blood-2013-02-486217. View

Cao Q, Qu M, Yang W, Wang D, Zhang M, Di S . Ischemia postconditioning preventing lung ischemia-reperfusion injury. Gene. 2014; 554(1):120-4. DOI: 10.1016/j.gene.2014.10.009. View

Yu H, Ma X, Ma X, Chen M, Chu Y, Wu L . Remote Limb Ischemic Postconditioning Protects Against Ischemic Stroke by Promoting Regulatory T Cells Thriving. J Am Heart Assoc. 2021; 10(22):e023077. PMC: 8751947. DOI: 10.1161/JAHA.121.023077. View

Xu Y, Li H, He X, Huang Y, Wang S, Wang L . Identification of the Key Role of NF-κB Signaling Pathway in the Treatment of Osteoarthritis With Bushen Zhuangjin Decoction, a Verification Based on Network Pharmacology Approach. Front Pharmacol. 2021; 12:637273. PMC: 8072665. DOI: 10.3389/fphar.2021.637273. View

Tohme S, Yazdani H, Sud V, Loughran P, Huang H, Zamora R . Computational Analysis Supports IL-17A as a Central Driver of Neutrophil Extracellular Trap-Mediated Injury in Liver Ischemia Reperfusion. J Immunol. 2018; 202(1):268-277. PMC: 6310076. DOI: 10.4049/jimmunol.1800454. View

Sharma A, Mulloy D, Le L, Laubach V . NADPH oxidase mediates synergistic effects of IL-17 and TNF-α on CXCL1 expression by epithelial cells after lung ischemia-reperfusion. Am J Physiol Lung Cell Mol Physiol. 2013; 306(1):L69-79. PMC: 3920214. DOI: 10.1152/ajplung.00205.2013. View

Wang T, Leng Y, Zhang Y, Kang Y, Xue X, Zhang Y . [Preventive effects of ischemic postconditioning and penehyclidine hydrochloride on gastric against ischemia-reperfusion injury in rats]. Zhonghua Yi Xue Za Zhi. 2011; 91(16):1130-5. View

Wilson A, Randall K, Pettitt J, Ellyard J, Blumenthal A, Enders A . Neutrophil extracellular traps and their histones promote Th17 cell differentiation directly via TLR2. Nat Commun. 2022; 13(1):528. PMC: 8792063. DOI: 10.1038/s41467-022-28172-4. View

10.

Wu J, Zhang F, Zheng X, Zhang J, Cao P, Sun Z . Identification of renal ischemia reperfusion injury subtypes and predictive strategies for delayed graft function and graft survival based on neutrophil extracellular trap-related genes. Front Immunol. 2022; 13:1047367. PMC: 9752097. DOI: 10.3389/fimmu.2022.1047367. View

11.

Brucken A, Derwall M, Bleilevens C, Stoppe C, Gotzenich A, Gaisa N . Brief inhalation of nitric oxide increases resuscitation success and improves 7-day-survival after cardiac arrest in rats: a randomized controlled animal study. Crit Care. 2015; 19:408. PMC: 4650396. DOI: 10.1186/s13054-015-1128-x. View

12.

Hirschhauser C, Lissoni A, Gorge P, Lampe P, Heger J, Schluter K . Connexin 43 phosphorylation by casein kinase 1 is essential for the cardioprotection by ischemic preconditioning. Basic Res Cardiol. 2021; 116(1):21. PMC: 7985055. DOI: 10.1007/s00395-021-00861-z. View

13.

Zhang H, Liu J, Zhou Y, Qu M, Wang Y, Guo K . Neutrophil extracellular traps mediate mA modification and regulates sepsis-associated acute lung injury by activating ferroptosis in alveolar epithelial cells. Int J Biol Sci. 2022; 18(8):3337-3357. PMC: 9134924. DOI: 10.7150/ijbs.69141. View

14.

Zhang Y, Chandra V, Riquelme Sanchez E, Dutta P, Quesada P, Rakoski A . Interleukin-17-induced neutrophil extracellular traps mediate resistance to checkpoint blockade in pancreatic cancer. J Exp Med. 2020; 217(12). PMC: 7953739. DOI: 10.1084/jem.20190354. View

15.

Wan R, Jiang J, Hu C, Chen X, Chen C, Zhao B . Neutrophil extracellular traps amplify neutrophil recruitment and inflammation in neutrophilic asthma by stimulating the airway epithelial cells to activate the TLR4/ NF-κB pathway and secrete chemokines. Aging (Albany NY). 2020; 12(17):16820-16836. PMC: 7521522. DOI: 10.18632/aging.103479. View

16.

Devaney J, Curley G, Hayes M, Masterson C, Ansari B, OBrien T . Inhibition of pulmonary nuclear factor kappa-B decreases the severity of acute Escherichia coli pneumonia but worsens prolonged pneumonia. Crit Care. 2013; 17(2):R82. PMC: 4056114. DOI: 10.1186/cc12696. View

17.

Zhang J, Dai Y, Wei C, Zhao X, Zhou Q, Xie L . DNase I improves corneal epithelial and nerve regeneration in diabetic mice. J Cell Mol Med. 2020; 24(8):4547-4556. PMC: 7176839. DOI: 10.1111/jcmm.15112. View

18.

Jovic S, Linge H, Shikhagaie M, Olin A, Lannefors L, Erjefalt J . The neutrophil-recruiting chemokine GCP-2/CXCL6 is expressed in cystic fibrosis airways and retains its functional properties after binding to extracellular DNA. Mucosal Immunol. 2015; 9(1):112-23. DOI: 10.1038/mi.2015.43. View

19.

Fei L, Jifeng F, Tiantian W, Yi H, Linghui P . Glycyrrhizin Ameliorate Ischemia Reperfusion Lung Injury through Downregulate TLR2 Signaling Cascade in Alveolar Macrophages. Front Pharmacol. 2017; 8:389. PMC: 5472719. DOI: 10.3389/fphar.2017.00389. View

20.

Qiu Z, Wang Y, Liu W, Li C, Zhao R, Long X . CircHIPK3 regulates the autophagy and apoptosis of hypoxia/reoxygenation-stimulated cardiomyocytes via the miR-20b-5p/ATG7 axis. Cell Death Discov. 2021; 7(1):64. PMC: 8024346. DOI: 10.1038/s41420-021-00448-6. View