Nitric Oxide Plays a Dual Role in Cardiorenal Syndrome in Vitro Model

Author(s)

Armentano, Giovana Marchini ; Pieretti, Joana Claudio ; Falconi, Carlos Alexandre ; Seabra, Amedea Barozzi ; Carneiro-Ramos, Marcela Sorelli

• Document type: Journal article
• Source: CELLULAR PHYSIOLOGY AND BIOCHEMISTRY; v. 58, n. 1, p. 16-pg., 2024-01-01.

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 2.9fold 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-alpha 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-1 beta 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. (c) 2024 The Author(s). Published by Cell Physiol Biochem Press GmbH&Co. KG (AU)

• FAPESP's process: 22/01124-4 - Impact of inflammatory response in cardiac and renal cells induced by accessory proteins of SARS-CoV-2: study of ORF 3a, ORF 6 and ORF 8 proteins
• Grantee: Giovana Marchini Armentano
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 22/00153-0 - Biophotonics applied to study of extracellular vesicles as connectors of kidney-heart crosstalk
• Grantee: Marcela Sorelli Carneiro Ramos
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/11077-0 - Cardiac alterations induced by renal inflammation models: participation of the Klotho/FGF-23 axis
• Grantee: Marcela Sorelli Carneiro Ramos
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/03646-2 - Impact of nanoplatforms allied to nitric oxide and chemotherapeutics on cytotoxicity and sensitization of resistant tumor cells
• Grantee: Joana Claudio Pieretti
• Support Opportunities: Scholarships in Brazil - Doctorate