İnflamatuar bağırsak hastalığı modellerinde TNF-alfa inhibitörlerine tamamlayıcı tedavi yaklaşımlarının geliştirilmesi
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Date
2023-08-10
Authors
Çamlıbel, Mine
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
İnflamatuar bağırsak hastalığı (IBD), ülseratif kolit (UC) ve Crohn hastalığı (CD) olarak iki ana formu içeren, kronik bir bozukluktur. Tümör Nekroz Faktörü’nün (TNF) fonksiyonunun baskılanmasına yönelik inhibitör ajan olan İnfliximab (IFX), IBD tedavisinde yaygın olarak kullanılmaktadır. IBD tedavilerindeki son dönemdeki gelişmelere rağmen, hastalarda tedaviye yanıt vermeme veya sonradan yanıtsızlık durumu görülebilmektedir. Ayrıca bu ilaçlar ciddi yan etkilere sahip olmakla birlikte neoplazi gelişim riskini de arttırabilmektedir. Bu nedenle, anti-inflamatuar aktivite gösteren doğal bileşiklerin, IBD için etkili terapötik stratejilerin geliştirilmesinde yeni bir yaklaşım oluşturabileceği düşünülmektedir. Mevcut tez çalışmasında doğal bir bileşik olan mirisetinin hem in-vitro IBD modeli hem de in-vivo UC modeli ve in-vivo CD modeli oluşturarak tek başına ve IFX ile kombin tedavisinin etkinliğinin araştırılması amaçlanmıştır. Bu doğrultuda hücre kültürü çalışmalarında LPS ve TNF-α ile indüklenen in-vitro IBD modelinde hücre proliferasyon testi gerçekleştirildi. Kolon kanseri hücre hattında kanserleşme süreci ile ilişkili yara iyileşmesi, koloni oluşumu gibi fonksiyonel analizler ve RT-PCR ile gen ekspresyon analizi gerçekleştirildi. Sıçanlar DSS ile indüklenerek in-vivo UC, TNBS ile indüklenerek in-vivo CD modeli oluşturularak klinik ve histopatolojik olarak değerlendirildi. Deney gruplarına ait kan ve kolon dokularında moleküler değişimler ELISA ve RT-PCR yöntemleriyle araştırıldı. Sonuç olarak mirisetinin in-vivo modellerde klinik parametreler üzerinde koruyucu etki gösterdiği, TNF-α salınımını ve inflamasyonla ilişkili NF-κB ve COX-2 genlerinin ekspresyonunu baskıladığı, Okludin ekspresyon seviyesi üzerinde koruyucu özelliği gösterilmiştir. Ayrıca kolon kanseri hücrelerinde kanserleşme sürecinde tümör agresifliğini de baskıladığı bulunmuştur. Mevcut bulgular mirisetinin IBD tedavisinde IFX ile birlikte yeni tedavi yaklaşımları oluşturma potansiyeline sahip olduğunu göstermektedir.
Inflammatory bowel disease (IBD) is a chronic disorder that includes two main forms, ulcerative colitis (UC) and Crohn's disease (CD). Infliximab (IFX), an inhibitory agent for the suppression of the Tumor Necrosis Factor (TNF) function, is widely used in the treatment of IBD. Despite recent advances in IBD treatments, patients may experience unresponsiveness or subsequent unresponsiveness to treatment. In addition, these drugs have serious side effects and may increase the risk of neoplasia development. Therefore, it is thought that natural compounds with anti-inflammatory activity may constitute a new approach in the development of effective therapeutic strategies for IBD. In the current thesis, it was aimed to investigate the efficacy of myricetin, a natural compound, both alone and in combination with IFX by creating both an in-vitro IBD model, an in-vivo UC model and an in-vivo CD model. Accordingly, cell proliferation test was performed in the in-vitro model of IBD induced by LPS and TNF-α in cell culture studies. Functional analyzes such as wound healing and colony formation related to cancerization process in colon cancer cell line and gene expression analysis by RT-PCR were performed. Rats were evaluated clinically and histopathologically by in-vivo UC induced with DSS and in-vivo CD model induced with TNBS. Molecular changes in blood and colon tissues of the experimental groups were investigated by ELISA and RT-PCR methods. As a result, it had been shown that myricetin had a protective effect on clinical parameters in in-vivo models, suppressed the expression of TNF-α and inflammation-related NF-kB and COX-2 genes, and had a protective effect on the expression level of Occludin. It had also been found to suppress tumor aggressiveness during the cancerization process in colon cancer cells. Current findings indicate that myricetin has the potential to create new therapeutic approaches in combination with IFX for the treatment of IBD.
Inflammatory bowel disease (IBD) is a chronic disorder that includes two main forms, ulcerative colitis (UC) and Crohn's disease (CD). Infliximab (IFX), an inhibitory agent for the suppression of the Tumor Necrosis Factor (TNF) function, is widely used in the treatment of IBD. Despite recent advances in IBD treatments, patients may experience unresponsiveness or subsequent unresponsiveness to treatment. In addition, these drugs have serious side effects and may increase the risk of neoplasia development. Therefore, it is thought that natural compounds with anti-inflammatory activity may constitute a new approach in the development of effective therapeutic strategies for IBD. In the current thesis, it was aimed to investigate the efficacy of myricetin, a natural compound, both alone and in combination with IFX by creating both an in-vitro IBD model, an in-vivo UC model and an in-vivo CD model. Accordingly, cell proliferation test was performed in the in-vitro model of IBD induced by LPS and TNF-α in cell culture studies. Functional analyzes such as wound healing and colony formation related to cancerization process in colon cancer cell line and gene expression analysis by RT-PCR were performed. Rats were evaluated clinically and histopathologically by in-vivo UC induced with DSS and in-vivo CD model induced with TNBS. Molecular changes in blood and colon tissues of the experimental groups were investigated by ELISA and RT-PCR methods. As a result, it had been shown that myricetin had a protective effect on clinical parameters in in-vivo models, suppressed the expression of TNF-α and inflammation-related NF-kB and COX-2 genes, and had a protective effect on the expression level of Occludin. It had also been found to suppress tumor aggressiveness during the cancerization process in colon cancer cells. Current findings indicate that myricetin has the potential to create new therapeutic approaches in combination with IFX for the treatment of IBD.
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Keywords
İnflamatuar bağırsak hastalığı, Mirisetin, TNF-α inhibitörü, İn-vivo IBD modelleri, Inflammatory bowel disease, Myricetin, TNF-α inhibitör, In-vivo IBD models
Citation
Çamlıbel, M. (2023). İnflamatuar bağırsak hastalığı modellerinde TNF-alfa inhibitörlerine tamamlayıcı tedavi yaklaşımlarının geliştirilmesi. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.