Determination of the expression profile of exenatide on 3D 3T3-L1 adipocytes by microarray analysis

Abstract

Obesity is a complex, multifactorial and important health problem worldwide. Current strategies for the treatment of obesity are quite limited. Therefore, the development of new drugs is very important. The aim of the study was to investigate the cytotoxicity of the GLP-1 agonist exenatide on 3 dimensional (3D) 3T3-L1 adipocytes through the whole genome profile. In this study, the apoptotic and lipolytic effect levels of exenatide on 3D adipocytes were also investigated. The cytotoxic activity of exenatide on 3T3-L1 adipocytes was determined by MTT method. Mouse gene microarray was used to screen for mRNAs. Apoptosis studies were performed by ELISA method. Lipolysis analysis was performed with the Lipolysis Colorimetric Assay Kit. In 3D adipose tissue modeling, exenatide application increased apoptosis level (p<0.01). Experiments with 3D 3T3-L1 adipocytes showed a high lipolytic effect in exenatide administration (p=0.000). In transcriptome analyzes performed to show the expression profiles of mRNAs in 3D adipocyte cells, a total of 3472 mRNAs were differentially expressed in the exenatide treated group compared to control adipocyte cells (p<0.05). The results obtained from this study may show that exenatide is effective in the treatment of obesity by regulating expression of various genes, adipocyte apoptosis and lipolysis activity.

Keywords

• Obesity
• microarray
• exenatide
• 3D cell culture
• 3T3-L1 cell

Project Number

220S827

Eksenatidin 3D 3T3-L1 Adipositleri Üzerindeki Ekspresyon Profilinin Mikrodizin Analizi ile Belirlenmesi

Abstract

Obezite karmaşık, çok faktörlü ve dünya genelinde önemli bir sağlık sorunudur. Obezite tedavisi için mevcut stratejiler oldukça kısıtlıdır. Bu nedenle yeni ilaç geliştirilmesi oldukça önemlidir. Çalışmanın amacı, GLP-1 agonisti olan eksenatidin 3 boyutlu (3D) 3T3-L1 adipositleri üzerindeki sitotoksisitesinin, tüm genom profili üzerinden araştırmaktır. Bu çalışmada, eksenatidin 3D adipositler üzerindeki apoptotik ve lipolitik etki düzeyleri de araştırıldı. Eksenatidin 3T3-L1 adipositleri üzerindeki sitotoksik aktivitesi MTT yöntemi ile belirlendi. mRNA' ları taramak için fare geni mikrodizisi kullanıldı. Apoptoz çalışmaları ELISA yöntemi ile yapıldı. Lipoliz analizi, Lipolysis Colorimetric Assay Kit ile gerçekleştirildi. 3D yağ doku modellemesinde eksenatid uygulaması apoptoz seviyesini arttırdı (p<0.01). 3D 3T3-L1 adipositleri ile yapılan deneyler, eksenatid uygulamasında lipolitik etkinin yüksek olduğunu gösterdi (p=0.000). 3D adiposit hücrelerinde mRNA'ların ekspresyon profillerini göstermek için yapılan transkriptom analizlerinde, eksenatid uygulanan grupta kontrol adiposit hücreleriyle kıyaslandığında toplam 3472 mRNA farklı şekilde ifade edildi (p<0.05). Bu çalışmadan elde edilen sonuçlar, eksenatidin, çeşitli genlerin anlatımını, adiposit apoptozunu ve lipoliz aktivitesini düzenleyerek obezite tedavisinde etkili olduğunu gösterebilir.

Keywords

• Obezite
• mikrodizin
• eksenatid
• 3D hücre kültürü
• 3T3-L1 hücresi

Supporting Institution

TÜBİTAK

Project Number

220S827

Thanks

Bu çalışma TÜBİTAK tarafından “220S827” kodlu proje ile desteklenmiştir. Bioink üretimi ile ilgili geri bildirimleri ve tavsiyeleri için Bugamed Teknoloji’ye (Türkiye) teşekkür ederiz.

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