Induction of oxidative stress by long-term treatment of live HEK293 cells with therapeutic concentration of lithium is associated with down-regulation of delta-opioid receptor amount and function

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00023752%3A_____%2F18%3A43919492" target="_blank" >RIV/00023752:_____/18:43919492 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/67985823:_____/18:00492456 RIV/68081715:_____/18:00492456

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0006295218302120?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0006295218302120?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.bcp.2018.06.004" target="_blank" >10.1016/j.bcp.2018.06.004</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Induction of oxidative stress by long-term treatment of live HEK293 cells with therapeutic concentration of lithium is associated with down-regulation of delta-opioid receptor amount and function

Popis výsledku v původním jazyce

The functional state of delta-opioid receptor signaling cascade in live cells exposed to a therapeutic concentration of lithium for a prolonged period of time (weeks) is not known because the previous studies of Li interference with OR were oriented to mu-OR only. The same applies to the analysis of the prolonged effect of Li on oxidative stress in context with delta-OR function. HEK293 cells stably expressing delta-OR were cultivated in the presence or absence of 1 mM LiCl for 7 or 21 days, homogenized and the post-nuclear (PNS) and plasma membrane (PM) fractions prepared from all four types of cells Level of delta-OR in PM was determined by specific radioligand [H-3]DADLE binding and immunoblot assays; the functional coupling between delta-OR and G proteins was determined as DADLE-stimulated high-affinity [S-35]GTP gamma S binding. In the whole cells, general oxidative stress was monitored by fluorescent dye 2&apos;,7&apos;-dichlorofluorescein diacetate (DCF) and results verified by analysis of PNS and isolated PM. Generation of 4-hydroxy-2-nonenal (4-HNE)-protein adducts and malondialdehyde (MDA) level were determined as products of lipid peroxidation. Li-treated cells exhibited the decreased amount of delta-OR. This was evidenced by both [H-3]DADLE binding and immunoblot assays. The delta-OR-G protein coupling efficiency was diminished Simultaneously, in Li-treated cells, the highly increased oxidative stress measured as DCF fluorescence intensity was noticed. Importantly, this result was detected in live cells as well as PNS and PM. Accordingly, production of 4-HNE-protein adducts and MDA was clearly increased in Li-treated cells. The general significance of our work lies in presentation of novel data indicating that prolonged exposure of live HEK293 cells to the therapeutic concentration of Li results in down-regulation of delta-OR protein level and attenuation of delta-OR function in parallel with increased oxidative stress and increased level of lipid peroxidation products.

Název v anglickém jazyce

Induction of oxidative stress by long-term treatment of live HEK293 cells with therapeutic concentration of lithium is associated with down-regulation of delta-opioid receptor amount and function

Popis výsledku anglicky

The functional state of delta-opioid receptor signaling cascade in live cells exposed to a therapeutic concentration of lithium for a prolonged period of time (weeks) is not known because the previous studies of Li interference with OR were oriented to mu-OR only. The same applies to the analysis of the prolonged effect of Li on oxidative stress in context with delta-OR function. HEK293 cells stably expressing delta-OR were cultivated in the presence or absence of 1 mM LiCl for 7 or 21 days, homogenized and the post-nuclear (PNS) and plasma membrane (PM) fractions prepared from all four types of cells Level of delta-OR in PM was determined by specific radioligand [H-3]DADLE binding and immunoblot assays; the functional coupling between delta-OR and G proteins was determined as DADLE-stimulated high-affinity [S-35]GTP gamma S binding. In the whole cells, general oxidative stress was monitored by fluorescent dye 2&apos;,7&apos;-dichlorofluorescein diacetate (DCF) and results verified by analysis of PNS and isolated PM. Generation of 4-hydroxy-2-nonenal (4-HNE)-protein adducts and malondialdehyde (MDA) level were determined as products of lipid peroxidation. Li-treated cells exhibited the decreased amount of delta-OR. This was evidenced by both [H-3]DADLE binding and immunoblot assays. The delta-OR-G protein coupling efficiency was diminished Simultaneously, in Li-treated cells, the highly increased oxidative stress measured as DCF fluorescence intensity was noticed. Importantly, this result was detected in live cells as well as PNS and PM. Accordingly, production of 4-HNE-protein adducts and MDA was clearly increased in Li-treated cells. The general significance of our work lies in presentation of novel data indicating that prolonged exposure of live HEK293 cells to the therapeutic concentration of Li results in down-regulation of delta-OR protein level and attenuation of delta-OR function in parallel with increased oxidative stress and increased level of lipid peroxidation products.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30104 - Pharmacology and pharmacy

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2018

Kód důvěrnosti údajů

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

Název periodika

Biochemical Pharmacology

ISSN

0006-2952

e-ISSN

—

Svazek periodika

154

Číslo periodika v rámci svazku

August

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

12

Strana od-do

452-463

Kód UT WoS článku

000441369600044

EID výsledku v databázi Scopus

2-s2.0-85048573684