İnsan retinal pigment epitel (Arpe-19) hücrelerinde paenol'ün miR-155 ve miR-210 aracılı hipoksi sinyalindeki rolü
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Dosyalar
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Hipoksi, oksijen homeostazının azalması durumu olarak tanımlanmaktadır. Hipoksi fizyolojik olarak özellikle embriyonik gelişim evresinde gerekli iken patolojik olarak meydana geldiğinde hücresel ölüme sebep olmaktadır. Retinal hipoksi; diyabetik retinopati, yaşa bağlı makula dejenerasyonu, prematüre retinopatisi, glokom, retinal vasküler hastalıklar gibi körlüğe neden olabilecek patolojilerin temel sebeplerindendir. Geleneksel Çin tıbbında kullanılmakta olan paeonol, Paeonia suffruticosa'nın kök kabuğundan izole edilen ana bileşen olup anti-enflamatuar, nöroprotektif, anti-tümör, antioksidan ve anti-anjiyogenik etkilere sahiptir. Bu çalışmada, DFX (Deferoxamine Mesylate Salt) ile hipoksi uygulaması yapılan insan retinal pigment epitel hücrelerinde (ARPE-19) paeonolün hipoksi ve anjiyogenez sinyali üzerine etkilerinin belirlenmesi amaçlanmıştır. Paeonolün hipoksi ile indüklenen ARPE-19 hücrelerinde hücre canlılığına olan etkileri MTT testi ile belirlenmiştir. Hipoksi ve anjiyogenez yolaklarının gen ve miRNA ifadeleri kantitatif gerçek zamanlı PCR, protein ifadeleri ise western blot analizi ile belirlenmiştir. Çalışma sonuçlarına göre paeonolün hipoksiye maruz kalan ARPE-19 hücrelerinde canlılığın artmasına neden olduğu belirlenmiştir. Buna ilaveten paeonol, hipoksi ile artan HIF-1?, HIF-1?, VEGF, VEGFR, MMP-3 ve MMP-9 geninde ve miR155 ve miR-210 ekspresyonlarında azalmalara neden olmuştur. Ayrıca HIF-1?, MMP-3 ve MMP-9 gen ekspresyonlarının kontrol seviyesine dönmesine de neden olmuştur. Çalışma sonuçlarımıza göre görmeden sorumlu fotoreseptörleri besleyen, koruyucu bariyer işlevi gören ve oksijen değişimlerine yüksek hassasiyeti olan retinal pigment epitel hücrelerinde meydana gelen hipoksinin paeonol aracılığıyla azaltılabileceği ve paeonolün terapötik bir rolünün olabileceği düşünülmektedir.
Hypoxia is defined as a state of reduced oxygen homeostasis. While hypoxia is physiologically essential especially in the embryonic developmental stage, when it occurs pathologically, it causes cellular death. Retinal hypoxia; it is one of the main causes of pathologies that may cause blindness such as diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, glaucoma, retinal vascular diseases. Paeonol which is used in traditional Chinese medicine, is the main component isolated from the root bark of Paeonia suffruticosa and has anti-inflammatory, neuroprotective, anti-tumor, antioxidant and anti-angiogenic effects. In this study, it was aimed to determine the effects of paeonol on hypoxia and angiogenesis signal in human retinal pigment epithelial cells (ARPE-19) treated with DFX (Deferoxamine Mesylate Salt). The effects of paeonol on cell viability in hypoxia-induced ARPE-19 cells were determined by MTT test. The gene and miRNA expressions of hypoxia and angiogenesis pathways were determined by quantitative real-time PCR, and protein expressions were determined by western blot analysis. According to the results of the study, it was determined that paeonol caused an increase in viability in ARPE-19 cells exposed to hypoxia. In addition, paeonol caused decreases in the HIF-1?, HIF-1?, VEGF, VEGFR, MMP-3 and MMP-9 gene and miR155 and miR-210 expressions, which is increased with hypoxia. Furthermore it was also caused the HIF-1?, MMP-3 and MMP-9 gene expressions return to the control level. According to results of the our study, it is thought that hypoxia that occurs in retinal pigment epithelial cells, which nourish the photoreceptors responsible for vision, function as a protective barrier and are highly sensitive to oxygen changes, can be reduced by paeonol. In addition, paeonol may have a therapeutic role.
Hypoxia is defined as a state of reduced oxygen homeostasis. While hypoxia is physiologically essential especially in the embryonic developmental stage, when it occurs pathologically, it causes cellular death. Retinal hypoxia; it is one of the main causes of pathologies that may cause blindness such as diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, glaucoma, retinal vascular diseases. Paeonol which is used in traditional Chinese medicine, is the main component isolated from the root bark of Paeonia suffruticosa and has anti-inflammatory, neuroprotective, anti-tumor, antioxidant and anti-angiogenic effects. In this study, it was aimed to determine the effects of paeonol on hypoxia and angiogenesis signal in human retinal pigment epithelial cells (ARPE-19) treated with DFX (Deferoxamine Mesylate Salt). The effects of paeonol on cell viability in hypoxia-induced ARPE-19 cells were determined by MTT test. The gene and miRNA expressions of hypoxia and angiogenesis pathways were determined by quantitative real-time PCR, and protein expressions were determined by western blot analysis. According to the results of the study, it was determined that paeonol caused an increase in viability in ARPE-19 cells exposed to hypoxia. In addition, paeonol caused decreases in the HIF-1?, HIF-1?, VEGF, VEGFR, MMP-3 and MMP-9 gene and miR155 and miR-210 expressions, which is increased with hypoxia. Furthermore it was also caused the HIF-1?, MMP-3 and MMP-9 gene expressions return to the control level. According to results of the our study, it is thought that hypoxia that occurs in retinal pigment epithelial cells, which nourish the photoreceptors responsible for vision, function as a protective barrier and are highly sensitive to oxygen changes, can be reduced by paeonol. In addition, paeonol may have a therapeutic role.
Açıklama
Anahtar Kelimeler
ARPE-19, DFX, miR-155, miR-210, miR-221, Hipoksi, Anjiyogenez, Paeonol, Hypoxia, Angiogenesis
