| Abstract | Aflatoksin B1 (AFB1), okratoksin A (OTA) i sterigmatocistin (STC) su sekundarni metaboliti filamentoznih gljivica roda Aspergillus i pokazuju toksične učinke na višim organizmima. Ta tri mikotoksina često dolaze u kombinaciji, a pri kroničnoj izloženosti u malim koncentracijama mogu dovesti do imunotoksičnih učinaka.
Površinski sloj kože (epidermis) je dio ljudskog tijela najizloženiji okolini, a sastavljen je najvećim djelom od keratinocita koji imaju važnu imunološku ulogu u koži, gdje su među najvećim producentima citokina.
Svrha ovog rada bila je odrediti utjecaj AFB1, OTA i STC, te njihovih binarnih kombinacija u koncentracijama od 1 i 10 μM, na otpuštanje citokina IL-1β, IL-6, IL-8, TNFα i IL-10 iz stanica humanih keratinocita (HaCaT stanice). Koncentracija navedenih citokina je određena primjenom enzimskog imunokemijskog testa u supernatantu stanica tretiranih navedenim koncentracijama mikotoksina kroz 24 sata. Rezultati su pokazali da netretirane HaCaT stanice ne proizvode IL-1β, TNFα i IL-10, dok IL-6 i IL-8 proizvode u koncentracijama od 507 pg/mL, odnosno 547 pg/mL. Nakon tretiranja mikotoksinima je uočen porast razina IL-6 i IL-8 u supernatantu od 123-297%, odnosno 131-228%. Pri ispitivanju binarnih kombinacija mikotoksina na izlučivanje citokina uočen je antagonistički učinak. Zaključujemo da pojedinačni mikotoksini AFB1, OTA i STC, te kombinacije AFB1+OTA, AFB1+STC i OTA+STC djeluju proupalno na HaCaT stanice povećavajući razinu IL-6 i IL-8, no taj je učinak za kombinacije manji u odnosu na djelovanje pojedinačnih mikotoksina, što je vjerojatno posljedica aditivnog i/ili sinergističkog citotoksičnog djelovanja tih kombinacija mikotoksina. |
| Abstract (english) | Aflatoxin B1 (AFB1), ochratoxin A (OTA) and sterigmatocystin (STC) are mycotoxins derived from filamentous fungi of genus Aspergillus. Co-occurrence of those three mycotoxins is often, and chronic exposure can lead to immunotoxicity. Outer skin layer (epidermis) is the most exposed part of human body. It is mostly made of keratinocytes which have the important role in skin immunology due to their cytokine production ability.
The aim of this study was to determine the effect of AFB1, OTA and STC, and also their binary combinations in concentrations of 1 and 10 μM on cytokine IL-1β, IL-6, IL-8, TNFα and IL-10 production in human keratinocytes (HaCaT cell line). The concentration of mentioned cytokines was measured in supernatant of cells after incubation for 24 hours with mycotoxins by using enzyme-linked immunosorbent assay. Results showed that untreated HaCaT cells do not express IL-1β, TNFα and IL-10, while concentrations of IL-6 and IL-8 were 507 pg/mL and 547 pg/mL, respectively. After treatment with mycotoxins increase of IL-6 and IL-8 was detected in the supernatant in relative amount of 123-297% and 131-228%, respectively. Treatment with binary combinations of mycotoxins resulted with antagonistic effect. Taking into account the increase of IL-6 and IL-8 concentrations we can conclude that AFB1, OTA and STC, as well as combinations AFB1+OTA, AFB1+STC and OTA+STC have pro-inflammatory effect on HaCaT cell line. Pro-inflammatory effects of mycotoxin combinations were less than pro-inflammatory effects of mycotoxins applied alone, which was probably consequence of additive or synergistic cytotoxic effects of mycotoxin binary mixtures. |
