Imunomodulatorna i antioksidativna svojstva 10- hidroksi-2-dekanoične kiseline
Immunomodulatory and antioxidant properties of 10-hydroxy-2-decanoic acid
Author
Mihajlović, Dušan A.Mentor
Čolić, MiodragCommittee members
Lukić, MiodragZelen, Ivanka

Vučević, Dragana
Metadata
Show full item recordAbstract
10-hidroksi-2-dekanoična kiselina (10-HDA) je jedinstvena i najzastupljenija
nezasićena masna kiselina matičnog mleča. Iako prvi naučni radovi o ovoj masnoj
kiselini datiraju još od pedesetih godina prošlog veka, dosadašnja istraživanja,
uglavnom na in vitro animalnim modelima, su pokazala njeno anti-inflamatorno
svojstvo. Međutim, veoma malo se zna o njenoj imunomodulatornoj i
antioksidativnoj ulozi. Stoga smo mi u ovoj studiji ispitivali efekat 10-HDA na
funkcije humanih ćelija imunskog sistema in vitro, kao i njeno potencijalno
antioksidativno svojstvo.
Dobijeni rezultati su pokazali da je 10-HDA u milimolarnim
koncentracijama (1mM i 2mM) inhibirala oksidativni prasak humanih
neutrofila periferne krvi stimulisanih forbol-12-miristat-13-acetatom (PMA),
N-formil-L-metionil-L-leucil-L-fenilalaninom (fMLP) ili opsonizovanim
zimozanom (opZy) u kulturi. Osim toga, ova masna kiselina je u istim
koncentracijama inhibirala kako spontanu, tako i PMA, fMLP i opZy indukovanu
apopt...ozu, ali i intenzitet PMA uzrokovane NEToze (engl. Neutrophil Extracellular
Traps) humanih neutrofila.
U kulturama humanih mononuklearnih ćelija periferne krvi (PBMNC)
stimulisanim fitohemaglutininom (PHA) 10-HDA je u koncentraciji od 500μM
ispoljila pretežno supresivni efekat na proliferaciju PBMNC. Ovaj proces je
bio praćen smanjenom produkcijom interleukina (IL)-2. U istoj koncentraciji, 10-
HDA je inhibirala sekreciju IL-1β i faktora nekroze tumora (TNF)-α.
Analizirajući citokinski T „helper“ (Th) profil u ovim kulturama pokazano je da
je 10-HDA, u koncentraciji od 500μM, inhibirala Th1 i Th2 polarizaciju
imunskog odgovora, a nije značajnije modulisala Th17 imunski odgovor.
Takođe, u ovoj studiji je ispitivan uticaj 10-HDA na maturaciju i funkciju
humanih dendritskih ćelija monocitnog porekla (MoDC). 10-HDA je, u
koncentraciji od 500μM, inhibirala maturaciju MoDC stimulisanih
lipopolisaharidom (LPS). Ovako stimulisane MoDC su produkovale manje
količine IL-12, IL-18 i TNF-α, i inhibirale Th1 i Th2 polarizaciju imunskog
odgovora, što je zaključeno na osnovu smanjenja koncentracije interferona (IFN)-γ
i IL-4 u supernatantima ko-kultura 10-HDA-tretiranih MoDC i alogenih CD4+ T
limfocita periferne krvi. Nasuprot ovim rezultatima, 10-HDA je, u niskoj
koncentraciji (50μM), stimulisala Th1, a inhibirala Th2 imunski odgovor.
Ispitujući antioksidativno svojstvo 10-HDA, na eksperimentalnim modelima
bez ćelija, u kojima se reaktivne kiseonične vrste produkuju čisto hemijskim
putem, pokazano je da ova masna kiselina, u milimolarnim koncentracijama (1mM-
4mM), ima sposobnost da „sakuplja“, odnosno reaguje sa superoksid anjon
radikalom i hipohlornom kiselinom i tako smanjuje njihovo reaktivno delovanje.
Na osnovu dobijenih rezultata može se zaključiti da 10-HDA ispoljava dozno-
zavisan efekat na odgovor humanih imunskih ćelija in vitro, pri čemu u visokim
koncentracijama (0,5mM-2mM) deluje inhibitorno, a u niskim koncentracijama
(50μM) blago stimulatorno. Takođe, pokazano je da 10-HDA u milimolarnim
koncentracijama (1mM-4mM) ispoljava direktno antioksidativno svojstvo.
10-hydroxy-2-decanoic acid (10-HDA) is unique and the most common fatty acid of
royal jelly. Although the first scientific papers about this fatty acid originate from fifties of
the last century, very little is known about its immunomodulatory and antioxidative
activity. Therefore, we examined the effects of 10-HDA on the function of human immune
cells in vitro, as well as its potential antioxidative properties.
The obtained results showed that 10-HDA at millimolar concentrations (1mM and
2mM) inhibited oxidative burst of human peripheral blood neutrophils stimulated with
phorbol-12-myristate-13-acetate (PMA), N-formyl-L-methionyl-L-leucyl-L-phenylalanine
(fMLP) or opsonized zymosan (opZy) and neutrophil extracellular traps formation induced
by PMA. In addition, 10-HDA at the same concentrations inhibited spontaneous apoptosis,
as well as PMA-, fMLP- and opZy-induced apoptosis of neutrophils.
10-HDA, at concentration of 500μM, inhibited the proliferation of
phytohaemaggluti...nin-activated human peripheral blood mononuclear cells (PBMNCs),
and the process was followed by a decrease in the production of interleukin 2 (IL-2). At
the same concentration, 10-HDA inhibited the production of IL-1β and tumor necrosis
factor-α (TNF-α) by stimulated PBMNCs. Regarding T helper (Th) cytokine profile,
higher concentration of 10-HDA (500μM), in contrast to the lower one (50μM), inhibited
both Th1 and Th2 responses, whereas the Th17 response was not significantly modulated,
as judged by the levels of interferon-γ (IFN-γ), IL-5 and IL-17A in culture supernatants,
respectively.
The effect of 10-HDA on maturation and functions of human monocyte-derived
dendritic cells (MoDCs) was also investigated. A lower dose (50 μM) stimulated T helper
(Th)1 and down-regulated Th2 immune responses, as judged by the levels of IFN-γ and
IL-4, respectively, in supernatants of 10-HDA-treated MoDCs cultivated with allogeneic
CD4+ T cells. In contrast, a higher dose of 10-HDA (500 μM) inhibited maturation of
lipopolysaccharide (LPS)-stimulated MoDCs. Such treated MoDCs produced lower levels
of IL-12, IL-18 and TNF-α and down-regulated both Th1 and Th2 immune responses.
Examining the antioxidant properties by using cell-free systems, we demonstrated that 10-
HDA at millimolar concentrations is a scavenger of superoxide anion radical and HOCl.
In conclusion, our results suggest that 10-HDA exerts different activity on human
immune cells, depending on applied concentrations, which is important when considering
its therapeutic immunomodulatory property. In addition, it was shown that 10-HDA, at
millimolar concentrations, exerts direct antioxidant properties.