TY - JOUR
T1 - Cytoprotective and Antioxidant Effects of Steen Solution on Human Lung Spheroids and Human Endothelial Cells
AU - Pagano, F.
AU - Nocella, C.
AU - Sciarretta, S.
AU - Fianchini, L.
AU - Siciliano, C.
AU - Mangino, G.
AU - Ibrahim, M.
AU - De Falco, E.
AU - Carnevale, R.
AU - Chimenti, I.
AU - Frati, G.
N1 - Funding Information:
The authors would like to thank “Fondazione Roma” for continuous support. This study was funded by “Sapienza” University grant to GF and by MIUR funding to IC (protocol 2015BN82FK).
Publisher Copyright:
© 2017 The American Society of Transplantation and the American Society of Transplant Surgeons
PY - 2017/7
Y1 - 2017/7
N2 - 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.
AB - 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.
KW - basic (laboratory) research/science
KW - cellular biology
KW - ischemia reperfusion injury (IRI)
KW - lung transplantation/pulmonology
KW - molecular biology
KW - organ perfusion and preservation
KW - signaling/signaling pathways
KW - tissue injury and repair
KW - translational research/science
UR - http://www.scopus.com/inward/record.url?scp=85018684636&partnerID=8YFLogxK
U2 - 10.1111/ajt.14278
DO - 10.1111/ajt.14278
M3 - Article
C2 - 28322021
AN - SCOPUS:85018684636
VL - 17
SP - 1885
EP - 1894
JO - American Journal of Transplantation
JF - American Journal of Transplantation
SN - 1600-6135
IS - 7
ER -