Proliferatif vitreoretinopati tedavisinde doğal molekül hispidulin kullanımının in vitro modelde araştırılması
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Dosyalar
Tarih
2022
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Yayıncı
Trakya Üniversitesi Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Proliferatif vitreoretinopati fibroselüler membranlar ve intraretinal fibrozisin gelişmesi ile oluşan ve yırtıklı retina dekolmanı cerrahisi sonrasında gelişen nüks retina dekolmanı olgularının en önemli nedenidir. Regmatojen retina dekolmanı meydana geldiğinde proliferatif vitreoretinopati başlatıcı iki ana olay meydana gelir. Bunlar retina hipoksisi ve kan retina bariyerinin bozulmasıdır. Yapılan çalışmalarda proliferatif vitreoretinopatili gözlerde alınan vitreus örneğinde normal gözlere orana dönüştürücü büyüme faktör-beta seviyesi daha yüksek bulunmuştur. Son yıllarda proliferatif vitreoretinopatinin patofizyolojisini anlama yönündeki bilgiler artmıştır. Proliferatif vitreoretinopatinin medikal tedavisinde en fazla çalışılmış maddelerden birisi bir kemoterapi ajanı olan 5-fluorourasildir. Fluorourasilin hayvan modellerinde proliferatif vitreoretinopati oranını azaltmada etkili olduğu gösterilmiştir. Proliferatif vitreoretinopatinin hücre biyolojisi ve moleküler mediyatörleri halen incelenmeye devam etmektedir. Hispidulin çeşitli bitkilerde bulunan antioksidatif, antiinflamatuar, antimutajenik, antiproliferatif ve antineoplastik özellikleri ile anjiogenezi baskılayan bir flavonoiddir. Bu araştırmada, hispidulin molekülünün proliferatif vitreoretinopati etiyolojisinde rol oyanayan proliferasyon, migrasyon ve anjiogenez yolağı üzerinde olası etkisinin belirlenmesi amaçlandı. Bu amaçla insan retina pigment epitel hücre serilerinden insan retinal pigment epitel hücre hattı- 19 hücrelerinde 10 ng/ml dönüştürücü büyümef faktör-b2 kullanılarak in vitro bir proliferatif vitreoretinopati modeli oluşturularak 48 saat boyunca 3 uM hispidulin ve 1uM hispidulin ile 5fluorourasil ile kombinasyonun etkinliğine bakıldı. Araştırmada proliferatif vitreoretinopati gelişiminden sorumlu mekanizmada önemli yer tutan hücre proliferasyonu ve epitelyal- mezenkimal geçiş sonucu gelişen hücre migrasyonunu inhibe edici etkinliği, olası terapötik potansiyelinin belirlenmesi ve tabanında yatan moleküler mekanizmaların aydınlatılması hedeflendi. Çalışmada hücre proliferasyon, hücre göçü deneyleri ve gerçek zamanlı polimeraz zincir reaksiyon yöntemi ile bu olaylarda rol alan bazı transkripsiyon faktörleri ile hedef genlerin kantitatif mesajci ribonükleik asit transkripsiyon değişimleri analiz edildi. Çalışmanın istatiksel analizinde gruplar arasındaki farklılıklar için varyans analizi (ANOVA) ve ardından çoklu karşılaştırmalar için de Duncan'ın çoklu karşılaştırma testi kullanıldı.
Proliferative vitreoretinopathy is characterized by fibrocellular membranes and intraretinal fibrosis and the most important cause of recurrent retinal detachment after rhegmatogenous retinal detachment surgery. When rheumatogenic retinal detachment occurs, two main proliferative vitreoretinopathy initiating events occur. These are retinal hypoxia and disruption of the blood retinal barrier. In previous studies, Transforming growth factor-beta level was found to be higher in vitreous samples taken from eyes with proliferative vitreoretinopathy compared to normal eyes. In recent years, knowledge about the pathophysiology of proliferative vitreoretinopathy has increased. One of the most studied substances in the medical treatment of proliferative vitreoretinopathy is 5-fluorouracil, which is a chemotherapic agent. Hispidulin is a flavonoid, which found in various plants, that suppresses angiogenesis with its antioxidative, anti-inflammatory, antimutagenic, antiproliferative and antineoplastic properties. In this study, it was aimed to determine the possible effect of hispidulin molecule on proliferation, migration and angiogenesis pathway that play a role in the etiology of proliferative vitreoretinopathy. For this purpose, an in vitro proliferative vitreoretinopathy model will be created using 10 ng/ml Transforming growth factor-b2 in A human retinal pigment epithelium cell line-19 cells from human retinal pigment epithelium cell lines and efficacy of 3uM hispidulin and combination of 1 uM hispidulin with 5-fluorouracil for 48 hours examinated. The aim of the study was to determine the therapeutic potential and to elucidate the underlying molecular mechanisms, inhibiting cell proliferation and cell migration resulting from epithelial-mesenchymal transition, which play an important role in the mechanism responsible for proliferative vitreoretinopathy development. In this study, cell proliferation cell migration assays and messenger ribonucleic acid transcription changes will be analyzed by realtime polimeraz chain reaxion method and the expressions of some proteins involved in these events will be analyzed by Western blot technique. In the statistical analysis of the study, analysis of variance (ANOVA) was used for the differences between the groups, followed by Duncan's multiple comparison test for multiple comparisons.
Proliferative vitreoretinopathy is characterized by fibrocellular membranes and intraretinal fibrosis and the most important cause of recurrent retinal detachment after rhegmatogenous retinal detachment surgery. When rheumatogenic retinal detachment occurs, two main proliferative vitreoretinopathy initiating events occur. These are retinal hypoxia and disruption of the blood retinal barrier. In previous studies, Transforming growth factor-beta level was found to be higher in vitreous samples taken from eyes with proliferative vitreoretinopathy compared to normal eyes. In recent years, knowledge about the pathophysiology of proliferative vitreoretinopathy has increased. One of the most studied substances in the medical treatment of proliferative vitreoretinopathy is 5-fluorouracil, which is a chemotherapic agent. Hispidulin is a flavonoid, which found in various plants, that suppresses angiogenesis with its antioxidative, anti-inflammatory, antimutagenic, antiproliferative and antineoplastic properties. In this study, it was aimed to determine the possible effect of hispidulin molecule on proliferation, migration and angiogenesis pathway that play a role in the etiology of proliferative vitreoretinopathy. For this purpose, an in vitro proliferative vitreoretinopathy model will be created using 10 ng/ml Transforming growth factor-b2 in A human retinal pigment epithelium cell line-19 cells from human retinal pigment epithelium cell lines and efficacy of 3uM hispidulin and combination of 1 uM hispidulin with 5-fluorouracil for 48 hours examinated. The aim of the study was to determine the therapeutic potential and to elucidate the underlying molecular mechanisms, inhibiting cell proliferation and cell migration resulting from epithelial-mesenchymal transition, which play an important role in the mechanism responsible for proliferative vitreoretinopathy development. In this study, cell proliferation cell migration assays and messenger ribonucleic acid transcription changes will be analyzed by realtime polimeraz chain reaxion method and the expressions of some proteins involved in these events will be analyzed by Western blot technique. In the statistical analysis of the study, analysis of variance (ANOVA) was used for the differences between the groups, followed by Duncan's multiple comparison test for multiple comparisons.
Açıklama
Anahtar Kelimeler
Epitelyal-mezenkimal transformasyon, Hispidulin, Proliferatif vitreoretinopati, Retina pigment epitel hücresi, Epithelial–mesenchymal transition, Proliferative vitreoretinopathy, Retina pigment epihelial cells
