Uridine Alleviates Sepsis-Induced Acute Lung Injury by Inhibiting Ferroptosis of Macrophage
Abstract
:1. Introduction
2. Results
2.1. Uridine Metabolism Homeostasis Was Disturbed in Sepsis-Induced ALI
2.2. Uridine Supplementation Alleviated Sepsis-Induced ALI and Inflammation of Lung Tissue
2.3. Uridine Inhibited Ferroptosis in Lung Tissue Dependent on Nrf2 Activation and ACSL4 Inhibition
2.4. Uridine Supplementation Restrained Inflammation and Oxidative Stress in THP-1 Cells
2.5. Uridine Suppressed Ferroptosis in THP-1 Cells via Activating Nrf2 and Inhibiting ACSL4 Expression
2.6. Nrf2 Knockdown Abrogated the Ferroptosis Resistance Effect of Uridine
2.7. Dysregulated Uridine Metabolism Was Confirmed in Patients with Sepsis
3. Discussion
4. Methods and Materials
4.1. Chemical and Reagents
4.2. Establishment of Sepsis-Induced Acute Lung Injury Model
4.3. BALF Collection, Processing and Determination of Protein Concentration
4.4. Lung Wet-to-Dry Weight Ratio Measurement
4.5. H&E Staining and Lung Injury Score Evaluation
4.6. Assay of MPO Activity
4.7. Quantification of Cytokines, Detection of Tissue Iron and Lipid Peroxidation Level
4.8. Cell Culture and Transfection
4.9. Macrophage Differentiation and Treatment
4.10. GSH Measurement
4.11. Cell Viability Assay
4.12. RNA Extraction and Real-Time Polymerase Chain Reaction
4.13. Western Blotting
4.14. ROS, Lipid Peroxidation and Mitochondrial Potential Measurement
4.15. Dataset Collection and Preprocessing
4.16. Bioinformatic Analysis and Visualization
4.17. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Species | Gene | Forward Primer | Reverse Primer |
---|---|---|---|
Mice | TNF-α | ACTGAACTTCGGGGTGATCGGT | TGGTTTGCTACGACGTGGGCTA |
Mice | IL-6 | CCCCAATTTCCAATGCTCTCC | CGCACTAGGTTTGCCGAGTA |
Mice | IL-1β | AATGAAGGAACGGAGGAGCC | CTCCAGCCAAGCTTCCTTGT |
Mice | GPX4 | GCTGGGAAATGCCATCAAAT | TCCTTCTCTATCACCTGGGGCT |
Mice | SLC7A11 | GAAATATCAGGTCATTGGTGGAGA | ATGCTCCTGCTTGAGTATGTCG |
Mice | HO1 | GCCAGCAACAAAGTGCAAGA | TAAGGACCCATCGGAGAAGC |
Mice | UPP1 | ATCCCAACATCTGTGCAGGC | ACCTGGCATGGTACAGCATC |
Mice | β-actin | GTCCACCGCAAATGCTTCTA | TGCTGTCACCTTCACCGTTC |
Human | TNF-α | TCCAGGCGGTGCTTGTTC | GCTTGTCACTCGGGGTTC |
Human | IL-6 | ATGAGGAGACTTGCCTGGTGAA | CTCTGGCTTGTTCCTCACTACTCTC |
Human | IL-1β | CGGGACTCACAGCAAAAAA | TTCAACACGCAGGACAGGT |
Human | GPX4 | TGAAGATCCAACCCAAGGGC | GACGGTGTCCAAACTTGGTG |
Human | SLC7A11 | GGCAGTTGCTGGGCTGATTTA | GATGACGAAGCCAATCCCTGT |
Human | HO1 | ATGCCCCAGGATTTGTCAGAG | GGAAGTAGACAGGGGCGAAGAC |
Human | UPP1 | CACCACTAGCAGACACAATTTCC | AATGCCCATACCATGACTGACAG |
Human | β-actin | CACCCAGCACAATGAAGATCAAGAT | CCAGTTTTTAAATCCTGAGTCAAGC |
Dataset Number | Platform | LPS Group/Sepsis | Control Group/Healthy | Annotation | Sample Type |
---|---|---|---|---|---|
GSE130936 | GPL339 | 6 | 4 | Affymetrix Mouse Expression 430A Array | Murine lung |
GSE15379 | GPL1261 | 3 | 3 | Affymetrix Mouse Genome 430 2.0 Array | Murine lung |
GSE2411 | GPL339 | 6 | 6 | Affymetrix Mouse Expression 430A Array | Murine lung |
GSE1871 | GPL1261 | 3 | 3 | Affymetrix Mouse Genome 430 2.0 Array | Murine lung |
GSE211210 | GPL24676 | 5 | 5 | Illumina NovaSeq 6000 (Homo sapiens) | Human Blood |
GSE137340 | GPL10558 | 45 | 12 | Illumina HumanHT-12 V4.0 expression beadchip | Human Blood |
GSE185263 | GPL16791 | 348 | 44 | Illumina HiSeq 2500 (Homo sapiens) | Human Blood |
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Lai, K.; Song, C.; Gao, M.; Deng, Y.; Lu, Z.; Li, N.; Geng, Q. Uridine Alleviates Sepsis-Induced Acute Lung Injury by Inhibiting Ferroptosis of Macrophage. Int. J. Mol. Sci. 2023, 24, 5093. https://doi.org/10.3390/ijms24065093
Lai K, Song C, Gao M, Deng Y, Lu Z, Li N, Geng Q. Uridine Alleviates Sepsis-Induced Acute Lung Injury by Inhibiting Ferroptosis of Macrophage. International Journal of Molecular Sciences. 2023; 24(6):5093. https://doi.org/10.3390/ijms24065093
Chicago/Turabian StyleLai, Kai, Congkuan Song, Minglang Gao, Yu Deng, Zilong Lu, Ning Li, and Qing Geng. 2023. "Uridine Alleviates Sepsis-Induced Acute Lung Injury by Inhibiting Ferroptosis of Macrophage" International Journal of Molecular Sciences 24, no. 6: 5093. https://doi.org/10.3390/ijms24065093