Cytoprotective and Antioxidant Effects of Steen Solution on Human Lung Spheroids and Human Endothelial Cells.
Respiratory diseases represent a major healthcare burden worldwide. Lung transplantation (LTx) is the "gold standard" for end-stage patients, strongly limited by shortage of available/suitable donor lungs. Normothermic ex vivo lung perfusion (EVLP) has significantly increased the number of lungs suitable for transplantation. Steen solution is used for EVLP, but the mechanisms involved in its beneficial properties remain to be clarified. We investigated the effects of Steen solution in an in vitro protocol of cold starvation and normothermic recovery on human lung spheroids, named pneumospheres (PSs), containing epithelial/basal cells, and on endothelial human umbilical vein endothelial cells (HUVEC). Steen solution significantly preserved the viability of PSs, reduced reactive oxygen species (ROS) release by PSs and HUVECs, decreased NADPH-oxidase (NOX) activity in PSs, and reduced inflammatory cytokines expression levels in HUVECs. Steen solution was able to specifically reduce NADPH oxidase 2 (NOX2) isoform activation, particularly in PSs, as detected by soluble-NOX2 peptide and p47-phosphorylation. Interestingly, a specific NOX2 inhibitor could partly mimic the pro-survival effect of Steen on PSs. We provide the first evidence that Steen solution can preserve lung epithelial/progenitor cells viability partially through NOX2 downregulation, and exert antioxidant effects on parenchymal cells, with consequent ROS reduction. These results suggest that NOX2 inhibition might be an additional strategy to reduce cellular damage during LTx procedures.
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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Pagano
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.|Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy.
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Nocella
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.|Department of AngioCardioNeurology, IRCCS Neuromed, Pozzilli, Italy.
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Sciarretta
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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Fianchini
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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Siciliano
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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Mangino
Department of Medical-Surgical Science and Translational Medicine, "Sapienza" University of Rome, Rome, Italy.
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Ibrahim
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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De Falco
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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Carnevale
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.
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Chimenti
Department of Medical Surgical Sciences and Biotechnology, "Sapienza" University of Rome, Rome, Italy.|Department of AngioCardioNeurology, IRCCS Neuromed, Pozzilli, Italy.
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Frati
D000293:Adolescent; D000328:Adult; D000975:Antioxidants / Q000494:pharmacology*; D002478:Cells, Cultured; D019610:Cytoprotection; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007552:Isotonic Solutions / Q000494:pharmacology*; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D000074662:NADPH Oxidase 2 / Q000378:metabolism; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015427:Reperfusion Injury / Q000517:prevention & control; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000378:metabolism; D055815:Young Adult
D000975:Antioxidants; D007552:Isotonic Solutions; D019188:Organ Preservation Solutions; D017382:Reactive Oxygen Species; D000074662:NADPH Oxidase 2
basic (laboratory) research/science; cellular biology; ischemia reperfusion injury (IRI); lung transplantation/pulmonology; molecular biology; organ perfusion and preservation; signaling/signaling pathways; tissue injury and repair; translational research/science