ULOGA JNK U PROINFLAMATORNOM DEJSTVU SARS-CoV-2 PROTEINA ORF3a
Sažetak
Uvod: Koronavirus-19 bolest (eng. Coronavirus disease 2019- COVID-19) je respiratorno oboljenje izazvano SARS-CoV-2 virusom (eng. Severe acute respiratory syndrome coronavirus 2 - SARS-CoV-2), koje je odnelo milione života za vreme pandemije 2020. godine i nastavlja da predstavlja značajan izazov za globalno zdravlje ljudi. SARS-CoV-2 protein ORF3a (eng. Open Reading Frame 3a - ORF3a) identifikovan je kao jedan od glavnih pokretača prekomernog inflamatornog odgovora povezanog sa težom kliničkom slikom i smrtnim ishodom usled COVID-19. c-Jun N-terminalna kinaza (eng. c-Jun N-terminal kinase - JNK), član familije mitogenom-aktiviranih protein kinaza (eng. Mitogen-activated protein kinases - MAPK) igra ključnu ulogu u citokinskoj ekspresiji i ćelijskom urođenom i stečenom inflamatornom odgovoru izazvanom različitim virusnim infekcijama.
Cilj rada: Cilj ovog istraživanja je bio da se ispita uloga JNK u ekspresiji gena za proinflamatorne citokine IL-1β i IL-8 na humanoj ćelijskoj liniji nesitnoćelijskog karcinoma pluća H460 koja eksprimira SARS-CoV-2 ORF3a.
Materijal i metode: H460 ćelijska linija transfektovana je DNK plazmidom koji kodira SARS-CoV-2 ORF3a. Aktivnost JNK u H460 ćelijama farmakološki je inhibirana tretmanom SP600215. Nivo proteina JNK određen je imunoblot metodom, a ekspresija proinflamatornih citokina IL-1β i IL-8 analizirana je na nivou iRNK primenom metode RT-qPCR. Statistička analiza je urađena Studentovim t-testom za nezavisne uzorke.
Rezultati: U H460 ćelijama transfektovanim plazmidom koji kodira SARS-CoV-2 ORF3a je utvrđen značajan porast nivoa fosforilacije JNK primenom imunoblot metode (p <0,05). RT-qPCR analiza je pokazala povećane nivoa ekspresije iRNK za proinflamatorne citokine IL-1β i IL-8 u transfektovanim ćelijama (p <0,05). Naknadna inhibicija JNK je značajno smanjila ekspresiju iRNK za navedene citokina (p <0,05), potvrđujući ulogu JNK u proinflamatornom dejstvu SARS-CoV-2 ORF3a.
Zaključak: Rezultati ove studije su pokazali da JNK igra značajnu ulogu u ekspresiji proinflamatornih citokina IL-1β i IL-8, indukovanih SARS-CoV-2 proteinom ORF3a. Selektivnom inhibicijom JNK bi se moglo smanjiti potencijalno štetno dejstvo prekomerne inflamacije izazvane proinflamatornim citokinima IL-1β i IL-8 kod teških slučajeva COVID-19.
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