Lipid membrane behavior of nitro-fatty acids and their loading into liposomes to activate Nrf2 pathway in RAW264.7 cells with impact on intracellular NO production

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15640%2F25%3A73633873" target="_blank" >RIV/61989592:15640/25:73633873 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61989592:15310/25:73633873 RIV/61989592:15110/25:73633873

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0009308425000337?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0009308425000337?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.chemphyslip.2025.105497" target="_blank" >10.1016/j.chemphyslip.2025.105497</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Lipid membrane behavior of nitro-fatty acids and their loading into liposomes to activate Nrf2 pathway in RAW264.7 cells with impact on intracellular NO production

Popis výsledku v původním jazyce

Nitro-fatty acids (NO2-FAs) are endogenous electrophilic signalling modulators, and some of them have been proposed as drug candidates. The main ones include nitro-oleic acid (NO2-OA) and other derivatives of unsaturated fatty acids such as nitro-linoleic acid (NO2-LA). In this study, we describe the behavior of 9/10-NO2-OA, 10-NO2-LA and the conjugated nitro-linoleic acid (9/12-NO2-cLA) in a model POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) membrane using molecular dynamics and selected experimental approaches. We showed that when loaded in liposomes, NO2-FAs undergo degradation (a decay reaction) to a very limited extent, in contrast to the free molecular form in an aqueous environment. This was confirmed by the electron paramagnetic resonance spectroscopic analysis of NO radical release. In general, NO2-FAs suppress membrane hydration, especially in the segment where the ester groups are located. Further, in the presence of NO2-FAs, there is increased membrane fluidity and a decrease in the degree of lipid order. These effects are greater for NO2-FAs than for their non-nitrated versions. The presence of a nitro group in close contact with the polar head groups was confirmed. This drives the tilt of the lipid chain which in turn induces membrane disorder. Protonated NO2-FAs penetrated more easily/deeper into the membrane structure than the dissociated forms and this makes the membrane bilayer surface more negatively charged based on zeta potential measurement. We also found that NO2-FAs incorporated into POPC liposomes retained their ability to activate the Nrf2 pathway. This was documented by an increased expression of heme oxygenase-1 at the level of mRNA, with a parallel decrease in protein levels of Keap1, in murine macrophage RAW264.7 cells. The NO2-FAs treatment resulted in an increase in intracellular NO level in vitro as determined by a genetically encoded G-geNOp sensor. This was confirmed at statistically significant level only for NO2-OA, not for NO2-LA or NO2-cLA. The results indicate that biologically relevant NO release may be strictly dependent on which NO2-FA is investigated. This study supports the hypothesis that NO2-FAs are distributed (co-localized) in cells and tissues in the lipid or aqueous phase, which affects whether they are mobile, stable, and thus biologically active.

Název v anglickém jazyce

Lipid membrane behavior of nitro-fatty acids and their loading into liposomes to activate Nrf2 pathway in RAW264.7 cells with impact on intracellular NO production

Popis výsledku anglicky

Nitro-fatty acids (NO2-FAs) are endogenous electrophilic signalling modulators, and some of them have been proposed as drug candidates. The main ones include nitro-oleic acid (NO2-OA) and other derivatives of unsaturated fatty acids such as nitro-linoleic acid (NO2-LA). In this study, we describe the behavior of 9/10-NO2-OA, 10-NO2-LA and the conjugated nitro-linoleic acid (9/12-NO2-cLA) in a model POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) membrane using molecular dynamics and selected experimental approaches. We showed that when loaded in liposomes, NO2-FAs undergo degradation (a decay reaction) to a very limited extent, in contrast to the free molecular form in an aqueous environment. This was confirmed by the electron paramagnetic resonance spectroscopic analysis of NO radical release. In general, NO2-FAs suppress membrane hydration, especially in the segment where the ester groups are located. Further, in the presence of NO2-FAs, there is increased membrane fluidity and a decrease in the degree of lipid order. These effects are greater for NO2-FAs than for their non-nitrated versions. The presence of a nitro group in close contact with the polar head groups was confirmed. This drives the tilt of the lipid chain which in turn induces membrane disorder. Protonated NO2-FAs penetrated more easily/deeper into the membrane structure than the dissociated forms and this makes the membrane bilayer surface more negatively charged based on zeta potential measurement. We also found that NO2-FAs incorporated into POPC liposomes retained their ability to activate the Nrf2 pathway. This was documented by an increased expression of heme oxygenase-1 at the level of mRNA, with a parallel decrease in protein levels of Keap1, in murine macrophage RAW264.7 cells. The NO2-FAs treatment resulted in an increase in intracellular NO level in vitro as determined by a genetically encoded G-geNOp sensor. This was confirmed at statistically significant level only for NO2-OA, not for NO2-LA or NO2-cLA. The results indicate that biologically relevant NO release may be strictly dependent on which NO2-FA is investigated. This study supports the hypothesis that NO2-FAs are distributed (co-localized) in cells and tissues in the lipid or aqueous phase, which affects whether they are mobile, stable, and thus biologically active.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA23-06051S" target="_blank" >GA23-06051S: Elektrofilní nitrované mastné kyseliny jako ochrana proti poškození krvetvorby způsobené ionizujícím zářením: ovlivnění dráhy NRF2</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

CHEMISTRY AND PHYSICS OF LIPIDS

ISSN

0009-3084

e-ISSN

1873-2941

Svazek periodika

270

Číslo periodika v rámci svazku

August 2025

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

14

Strana od-do

105497

Kód UT WoS článku

001501718000002

EID výsledku v databázi Scopus

2-s2.0-105006687897