INSL3, N9 mikroglia hücrelerinde LPS ile indüklenen enflamasyonu baskılar

Yıl 2024, Cilt: 49 Sayı: 2, 489 - 496, 30.06.2024

Dilek Şaker , Gülfidan Coşkun , Sait Polat

https://doi.org/10.17826/cumj.1455491

Öz

Amaç: İnsülin Benzeri Peptid 3 (INSL3) reseptörünü de içeren G-protein kaplı reseptör (GPCR) ailesi, inflamasyonda Nükleer Faktör kappa B (NF-κB) aracılı yolakta yer almaktadır. Bu bakımdan INSL3'ün NF-κB yolu üzerinden inflamasyonda rol oynadığı düşünülebilir. Bu çalışmada INSL3'ün, lipopolisakkarit (LPS) ile indüklenen N9 mikroglia hücre hattında inflamasyon ve hücre canlılığı üzerindeki etkisini araştırdık.
Gereç ve Yöntem: N9 mikroglial hücreleri, 2 saat boyunca INSL3 ile ön işleme tabi tutuldu, ardından 6 saat boyunca LPS ile işleme tabi tutuldu. Hücre canlılığı WST-8 analizi ile belirlendi. İnterlökin-1β (IL-1β), Tümör nekroz faktörü (TNF)-α ve NF-κB düzeyleri de immünhistokimyasal yöntemlerle değerlendirildi.
Bulgular: LPS grubundaki hücrelerde morfolojide dejeneratif değişiklikler ve hücre canlılığında azalma görüldü. INSL3+LPS (1.21±0.06) grubunda hücrelerin genel görünümü ve canlılığı LPS (0.61±0.05) grubuna göre kontrol grubuna (1.92±0.04) daha benzerdi. INSL3'ün LPS kaynaklı IL-1β, TNF-α ve NF-κB seviyelerindeki artışı önlediği ve hücre ölümünü azalttığı belirlendi.
Sonuç: INSL3'ün inflamasyonu baskılayarak hücresel iyileşmeyi desteklediği ve inflamasyonu azaltan terapötik bir ajan olarak değerlendirilebileceği sonucuna varıldı.

Anahtar Kelimeler

Inflamasyon, insülin benzeri peptid 3, lipopolisakkarit, mikroglia

INSL3 suppresses LPS-induced inflammation in N9 microglia cells

Öz

Purpose: The G-protein coated receptor (GPCR) family, including the Insulin-Like Peptide 3 (INSL3) receptor, is involved in the Nuclear Factor kappa B (NF-κB)-mediated pathway in inflammation. In this regard, it can be thought that INSL3 plays a role in inflammation via the NF-κB pathway. In this study, we investigated the effect of INSL3 on inflammation and cell viability in the lipopolysaccharide (LPS)-induced N9 microglia cell line.
Materials and Methods: N9 microglial cells were pretreated with INSL3 for 2 hours, and then treated with LPS for 6 hours. Cell viability was identified by WST-8 assay. Immunostaining was performed to evaluate the levels of Interleukin-1β (IL-1β), Tumor necrosis factor (TNF)-α, and NF-κB.
Results: The cells in the LPS group showed degenerative changes in morphology and decreased cell viability. In the INSL3+LPS group (1.21±0.06), the general appearance and viability of the cells were more similar to the control group (1.92±0.04) compared to the LPS group (0.61±0.05). It was determined that INSL3 prevented the LPS-induced increase in IL-1β, TNF-α, and NF-κB levels and decreased cell death.
Conclusion: INSL3 suppresses inflammation and thus promotes cellular healing and can be considered a therapeutic agent that reduces inflammation.

Anahtar Kelimeler

inflammation, insulin-like peptide 3, lipopolysaccharide, microglia

Kaynakça

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