Damla SETENCİ, Merve Nur Sultan YILDIRIM, Ahmet CUMAOĞLU

LİPOPOLİSAKKARİT/NİGERİSİN İLE MAKROFAJ (RAW 264.7) HÜCRELERİNDE NLRP3 İNFLAMAZOM AKTİVASYONU: PSORALİDİN’İN REGÜLATÖR ETKİSİ

Prenile edilmiş bir komestan olan psoralidin'in anti-inflamatuvar etkisi bildirilmiştir, ancak inflamazom aktivasyonu ve piroptozis ile ilişkili sitokinler üzerindeki düzenleyici etkileri açık değildir. Bu çalışmanın amacı, psoralidinin Lipopolisakkarit (LPS)/Nigerisinile uyarılan RAW 264.7 makrofajlarında inflamazom aktivasyonu ve piroptozis ile ilişkili sitokinlerin ifadelenmesi üzerindeki düzenleyici rolünü araştırmaktı. Psoralidinin inflamazom aktivasyonu üzerindeki düzenleyici mekanizması, NOD (Nükleotid bağlama ve oligomerizasyon alanı) benzeri reseptör proteini 3 (NLRP3), adaptör protein apoptozla ilişkili benek benzeri protein (ASC) ve Kaspaz-1 için Western blot kullanılarak değerlendirildi. Piroptozis ile ilişkili sitokinler, interlökin-1β(IL-1β)ve interlökin-18 (IL-18) için RT-PCR kullanılarak değerlendirildi. Psoralidin'in sitotoksik olmayan konsantrasyonu (5.0μM) LPS-Nigerisin ile uyarılan inflamazom aktivasyonunu önemli ölçüde inhibe etti. Ek olarak, psoralidin ile ön tedavi, LPS/Nigerisin kaynaklı IL-1β ve IL18 üretimini bastırdı. Sonuçlarımız, Psoralidin'in RAW 264.7 makrofajlarında inflamazom aktivasyonu ve piroptozis ile ilişkili sitokin üretimi üzerindeki düzenleyici etkilerinin, NLRP3 inflamazom aktivasyonunun baskılanması ve piroptoz ile ilgili sitokinlerin salınımının inhibisyonu ile ilişkili olduğunu göstermektedir. Bu sonuçlara göre, Psoralidin, inflamatuvar hastalıkların önlenmesi veya inflamasyon tedavisinin desteklenmesi için potansiyel bir terapötik aday olarak düşünülebilir.

Anahtar Kelimeler:

Inflamazomlar, Lipopolisakkaritler, Piroptozis, Sitokinler

NLRP3 INFLAMMASOME ACTIVATION IN MACROPHAGE (RAW 264.7) CELLS BY LIPOPOLISACCARIDE/NIGERICIN: REGULATORY EFFECT OF PSORALIDIN

Psoralidin, a prenylated coumestan, has been reported its anti-inflammatory effect, but the regulatory effects on inflammasome activation and pyroptosis-related cytokines are not clear. The aim of this study was to investigate the regulatory role of psoralidin on inflammasome activation and expression of pyroptosis-related cytokines in Lipopolysaccharide (LPS)/Nigericin-stimulated RAW 264.7 macrophages. Regulatory mechanism of psoralidin on inflammasome activation was evaluated using Western blotting for NLRP3, adaptor protein apoptosis-associated speckle-like protein (ASC)and Caspase-1. Pyroptosis-related cytokines were evaluated using RT-PCR for interleukin-1β (IL-1β)and interleukin-18 (IL18).Non-cytotoxic concentration of Psoralidin (5.0µM) significantly inhibited LPS/Nigericin induced inflamasome activation. In addition, pretreatment with psoralidin suppressed the LPS/Nigericin-induced production of IL-1β and IL18. Our results indicate that the regulatory effects of psoralidin on inflammasome activation and pyroptosis related cytokine production in RAW 264.7 macrophages are associated with suppression of NLRP3inflammasome activation and inhibition of the release of pyroptosis related cytokines.According to these results, Psoralidine may be considered as a potential therapeutic candidate for the prevention of inflammatory diseases or to support the treatment of inflammation.

Keywords:

Inflammasomes, Lipopolysaccharides, Pyroptosis, Cytokines,

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