Changes in Polarity and Regeneration-Related Gene Expression in In Vitro Bone Marrow Mesenchymal Stem Cells in a Rheumatoid Arthritis Injury Model and Pharmacological Modulation

Abstract

Purpose: The aim of this study was to investigate the changes in polarity and regeneration at the gene-protein level with pharmacological modulation of rheumatoid arthritis (RA) disease, which occurs as a result of chronic, inflammation-induced tissue erosion whose pathogenesis steps are well understood. Materials and Methods: To investigate the damage caused by the pathogenetic steps of the disease to intracellular polarity and regeneration, bone marrow mesenchymal stem cells subjected to injury mimicry using IL-1β and IL-6 were treated with a combination of the IL-1β antagonist antibody canakinumab and the IL-6 antagonist antibody tocilizumab. The effects on cell viability, cytotoxicity, and the expression of genes related to intracellular polarity and regeneration were examined. Results: As a result of the analyses, as expected, IL-1β and IL-6 caused a reduction in viability and an acceleration in cytotoxicity in human bone marrow stem cells imitating RA damage. In addition, in gene expression analyses, low significant changes were observed in the expression of genes consisted in polarity pathways, but significant changes were found in the expression of genes take place in regeneration mechanisms. In addition, the application of antagonist agents reversed this situation and limited normalization and even changes that increased the viability of the cells in viability tests were observed. Conclusion: In this case, in the RA-like pathogenesis model obtained after injury mimicry, it is thought that the inflammation occurring during the disease's developmental stages causes stem cells to lose their properties such as adhesion and navigation, thereby affecting the efficiency of the regenerative properties they maintain.

Keywords

• arthritis
• bone marrow mesenchymal stem cell
• ınterleukin
• monoclonal antibody

Romatoid Artrit Yaralanma Modelinde In Vitro Kemik İliği Mezenkimal Kök Hücrelerinde Polarite ve Rejenerasyonla İlişkili Gen İfadesindeki Değişiklikler ve Farmakolojik Modülasyonu

Abstract

Amaç: Bu çalışmanın hedefi patogenez basamakları iyi anlaşılmış kronik, inflamasyon sebebiyle dokusal erozyonun sonucunda oluşan Romatoid Artirit (RA) hastalığının farmakolojik modülasyon ile gen-protein düzeyinde polarite ve rejenerasyondaki değişimleri incelenmiştir. Araçlar ve Yöntem: Hastalığın patogenez basamaklarının hücre içi polarite ve rejenerasyona verdiği zararı araştırabilmek için Il-1β ve IL-6 ile hasar taklidi yapılmış kemik iliği mezenkimal kök hücrelerinde IL-1β antagonist antikoru canakinumab ve IL-6 antagonist antikoru tocilizumab kombinasyon halinde uygulanmasının hücre canlılığı, sitotoksisite ve hücre içi polarite ve rejenerasyon ile ilgili genlerin ifadesi incelenmiştir. Bulgular: Analizler sonucunda IL-1β ve IL-6 beklendiği üzere RA hasar taklidi yapılan insan kemik iliği kök hücrelerinde canlılığının azalmasına sitotoksisitenin aynı oranda artmasına sebep olmuştur. Bunun yanında gen ifade analizlerinde polarite yolaklarında görevli genlerin ifadelenmesinde düşük oranda anlamlı değişiklik görülmüş ancak rejenerasyon ile ilgili genlerin ifadelenmelerinde anlamlı değişikliklere rastlanmıştır. Bunun yanında antagonist ajanların uygulanması bu durumu tersine çevirmiş sınırlı seviyede normalleşme hatta hücrelerin canlılık testlerinde canlılığı artıran değişimler gözlemlenmiştir. Sonuç: Bu durumda hasar taklidi sonrası elde edilen RA benzeri patogenez modelinde hastalığın gelişim basamaklarında gerçekleşen inflamasyonun etkisiyle kök hücrelerin adherasyon, yön bulma, gibi özelliklerini kaybetmeleri, korudukları rejenerasyon özellik verimliliğini etkilemekte olduğu düşünülmüştür.

Keywords

• artrit
• ınterlökin
• kemik iliği mezenkimal kök hücresi
• monoklonal antikor

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