Effects of Metformin on TNF- Release in Lipopolysaccharide-Induced Monocytes in Rats

Emre Gedikli; Mesut Parlak; Ahmet Serdar Soydan

doi:10.7197/cmj.1413260

TR EN

Sıçanlarda Lipopolisakkarid ile İndüklenen Monositlerde Metforminin TNF- Salınımı Üzerine Etkileri

Öz

Amaç: İmmün sistem yanıtının bir sonucu olan inflamasyon, pek çok hastalıkta ortaya çıkan bir tablodur (ateroskleroz, hipertansiyon, diyabet, kanser, romatoid artrit gibi). Diyabet tedavisinde kullanılan metforminin, kan şekerini düzenleyici etkisinden bağımsız olarak, antienflamatuvar etkisininde olduğu bulunmuştur. Ancak etki mekanizması tam olarak açıklanamamıştır. Bu çalışmada, metforminin, lipopolisakkarit (LPS) ile inkübe edilmiş sıçan monositlerinden proinflamatuar sitokinler olan TNF-α ve IL-6 sentez ve salınımı üzerindeki etkisi araştırıldı. Yöntem: Erkek 8-12 haftalık sağlıklı sıçanlardan (n=5, 200-250 g), genel anestezi (Xylazin 3 mg/kg + Ketamin 90 mg/kg) altında kardiyak puncture yöntemiyle 5 ml kan alındı ve steril EDTA’lı tüplere konuldu. Santrifuj (ficoll 3ml, 400xg, 30dk) yardımıyla mononükleer hücreler ayrıştırıldı. Hücreler RPMI 1640 media ile sulandırıldı (3.3±0.2x105 /ml). Hücreler önce metformin (2.5 μM, 25 μM, 250 μM) ile 2,5 saat inkübe edildi. Bu sürenin sonunda LPS (100 ng/ml, 1μg/ml) eklenerek 5 saat daha inkübe edildi. Daha sonra santrifuj (400xg, 20dk.) yapıldı, süpernatantlar -80oC’de, ELISA yöntemi ile TNF-α ve IL-6 düzeyleri ölçülene kadar saklandı. Bulgular: Hücrelerin deneyler sonunda canlılık oranları %98’in üzerindeydi. TNF-α miktarları arasındaki fark, LPS 100 ng/ml ile, LPS + metformin (2.5μM, 25μM, 250μM) grupları karşılaştırıldığında (2179±359 ve 1613±437, 2915±572, 6059±948 pg/ml), istatistiksel olarak anlamlı bulunmadı (p>0.05). LPS 1μgr/ml ile, LPS+metformin (2,5μM, 25μM, 250μM) grupları karşılaştırıldığında (1752±553 ve 3023±745, 2344±598, 6238±841 pg/ml), 2.5μM (p0.05). IL-6 miktarları ELISA yöntemi ile ölçülecek miktarda bulunmadı. Bu konuda yapılan diğer çalışmalardan elde edilen sonuçlara göre, metforminin proinflamatuar sitokinlerin sentezini inhibe ettiği bulunmuştur. Sonuç: Bu çalışmada, metformin, LPS ile indüklenen sıçan mononükleer hücrelerinden, TNF-α sentez ve salınımını, azaltmaktan ziyade artırmaya yönelik bir etki gösterdi.

Anahtar Kelimeler

Effects of Metformin on TNF- Release in Lipopolysaccharide-Induced Monocytes in Rats

Abstract

Objective: Inflammation which is a response of immune system was demonstrated in many disorders such as atherosclerosis, hypertension, diabetes, cancer and rheumatoid arthritis. Metformin, an oral antidiabetic drug, has anti-inflammatory effect apart from blood glucose regulatory effect. However, the mechanism of its anti-inflammatory effect is not clearly understood. In this study, the effect of metformin on the release of cytokines (TNF-α and IL-6) from LPS stimulated rat mononuclear blood cells was investigated. Methods: Blood samples (5ml) were taken from healthy, male, 8-12 weeks old rats (n=5, 200-250g) through cardiac puncture under general anesthesia into sterile EDTA containing tubes. Monocytes were separated by centrifugation and were resuspended in RPMI 1640 media (3.3±0.2x105 /ml). Cells were then incubated with metformin (2.5 μM, 25 μM, 250 μM) for 2,5 hours followed by addition of LPS (100 ng/ml, 1μg/ml) for further 5 hours. After centrifugation, the supernatant was taken and TNF-α level was measured by ELISA. Results: There was no statistically significant change in the amounts of TNF-α in the LPS + metformin groups compared to the 100 ng/ml LPS group (p>0.05). In LPS+metformin groups, compared to 1 μg/ml LPS, 2.5 μM and 250 μM metformin significantly increased TNF-α levels (p<0.05), while 25 μM metformin did not make a significant difference (p>0.05). The amount of IL-6 was not within measurable range in this study. Conclusion: In summary, metformin increased the amount of released TNF-α rather than decreased in our study.

Keywords

Kaynakça

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Ayrıntılar

Birincil Dil

İngilizce

Konular

Tıp Eğitimi

Bölüm

Araştırma Makalesi

Yazarlar

Emre Gedikli ^*
0000-0001-5593-2073
Türkiye

Mesut Parlak
0000-0001-9692-8396
Türkiye

Ahmet Serdar Soydan
0009-0006-1121-2967
Türkiye

Yayımlanma Tarihi

29 Mart 2024

Gönderilme Tarihi

2 Ocak 2024

Kabul Tarihi

22 Ocak 2024

Yayımlandığı Sayı

Yıl 2024 Cilt: 46 Sayı: 1

DOI

https://doi.org/10.7197/cmj.1413260

IZ

https://izlik.org/JA55SC23SD

Kaynak Göster

RIS / Bibtex

AMA

1.Gedikli E, Parlak M, Soydan AS. Effects of Metformin on TNF- Release in Lipopolysaccharide-Induced Monocytes in Rats. CMJ. 2024;46(1):29-34. doi:10.7197/cmj.1413260

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