Nitric Oxide Plays a Dual Role in Cardiorenal Syndrome in Vitro Model
Background/Aims:Nitric oxide (NO) plays a dual role, acting as both an oxidant and a reducer, with various effects depending on its concentration and environment. Acute kidney injury’s (AKI) pathogenesis observed in cardiorenal syndrome 3 (CRS 3) involves inflammatory responses and the production of reactive oxygen and nitrogen species. However, the role of NO on the development of CRS 3 is still not completely understood. The study aimed to mimic CRS 3 in vitro and investigate NO signaling and inflammatory molecules. Methods:Thus, HEK293 cells were submitted to normoxia (NX) or hypoxia (HX) protocols for 16 h followed by 3 h of reoxygenation, treated or not with L -NAME. Conditionate medium by HEK293 was transferred to H9c2 for 24 h. Cellular viability was evaluated by MTT assay, real time PCR was used to analyze gene expression and NO content were evaluated in the intra and extracellular medium by amperimetry. Results:Carbonic anhydrase 9 (CA9) expression increased 2.9-fold after hypoxia. Hypoxia reduced 18 % cell viability in HEK293 that was restored by L -NAME treatment. The sum of nitrite (NO2-) and S-nitrosothiol (S-NO) fractions in HEK293 cells showed a substantial decrease on NO intracellular content (38 %). Both IL-6 and IL-10 decreased in all groups compared to NX cells. Besides TNF-α and Bax/Bcl2 ratio increased in hypoxia (approximately 120-fold and 600-fold, respectively) and L -NAME restored this effect. Regarding H9c2 cells, the S-NO fractions showed a substantial decrease in extracellular content after HX (17%) that was not restored by L -NAME. IL-1β decreases in cardiac cells treated with conditioned medium from HX/L-NAME. Conclusion:In conclusion this study highlights the complex interplay of NO and inflammatory factors in hypoxia-induced renal and cardiac cell responses, with potential implications for cardiorenal syndrome.
Citação
@online{armentano,_giovana2024,
author = {Armentano, Giovana, Marchini and Joana Claudio , Pieretti
and Carlos Alexandre , Falconi and Seabra, Amedea, Barozzi and
Marcela Sorelli , Carneiro-Ramos},
title = {Nitric Oxide Plays a Dual Role in Cardiorenal Syndrome in
Vitro Model},
volume = {58},
number = {1},
date = {2024-01-22},
doi = {10.33594/000000681},
langid = {pt-BR},
abstract = {Background/Aims:Nitric oxide (NO) plays a dual role,
acting as both an oxidant and a reducer, with various effects
depending on its concentration and environment. Acute kidney
injury’s (AKI) pathogenesis observed in cardiorenal syndrome 3 (CRS
3) involves inflammatory responses and the production of reactive
oxygen and nitrogen species. However, the role of NO on the
development of CRS 3 is still not completely understood. The study
aimed to mimic CRS 3 in vitro and investigate NO signaling and
inflammatory molecules. Methods:Thus, HEK293 cells were submitted to
normoxia (NX) or hypoxia (HX) protocols for 16 h followed by 3 h of
reoxygenation, treated or not with L -NAME. Conditionate medium by
HEK293 was transferred to H9c2 for 24 h. Cellular viability was
evaluated by MTT assay, real time PCR was used to analyze gene
expression and NO content were evaluated in the intra and
extracellular medium by amperimetry. Results:Carbonic anhydrase 9
(CA9) expression increased 2.9-fold after hypoxia. Hypoxia reduced
18 \% cell viability in HEK293 that was restored by L -NAME
treatment. The sum of nitrite (NO2-) and S-nitrosothiol (S-NO)
fractions in HEK293 cells showed a substantial decrease on NO
intracellular content (38 \%). Both IL-6 and IL-10 decreased in all
groups compared to NX cells. Besides TNF-α and Bax/Bcl2 ratio
increased in hypoxia (approximately 120-fold and 600-fold,
respectively) and L -NAME restored this effect. Regarding H9c2
cells, the S-NO fractions showed a substantial decrease in
extracellular content after HX (17\%) that was not restored by L
-NAME. IL-1β decreases in cardiac cells treated with conditioned
medium from HX/L-NAME. Conclusion:In conclusion this study
highlights the complex interplay of NO and inflammatory factors in
hypoxia-induced renal and cardiac cell responses, with potential
implications for cardiorenal syndrome.}
}