Sıçan in vivo prematür ovaryan yetmezliği modelinde ovaryan mekanik uyarılmanın foliküler aktivasyona etkisi; intraovaryan iğne ve plateletten zengin plazma (PRP) enjeksiyonunun karşılaştırmalı analizi
Date
2024-07-17
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Publisher
Bursa Uludağ Üniversitesi
Abstract
Tez çalışmasının amacı, infertilite nedenlerinden biri olan prematür ovaryan yetmezlik (POI) olgularında, kontrollü ovaryan hiperstimülasyon (KOH) protokolü öncesi intraovaryan mekanik uyarma ve intraovaryan PRP enjeksiyonu uygulamasının hippo sinyal yolağı üzerinden ovaryan foliküllerin gelişimi üzerindeki etkisinin ve mekanizmasının karşılaştırmalı olarak değerlendirilmesidir. Deneysel POI modeli oluşturmak, intraovaryan enjeksiyonlar ve KOH protokolü uygulamak amacıyla 45 adet dişi Sprague-Dawley cinsi sıçan 7 farklı gruba ayrıldı. Denekler üzerinde 15 gün boyunca subkutan olarak tek doz 4-VinilsikloheksenDiepoksit (VCD, 160 mg/kg/gün) enjeksiyonu ile POI modeli oluşturuldu. VCD enjeksiyonları ardından intraovaryan olarak; cerrahi yolla açma kapama, boş iğne ile mekanik uyarma, PRP ve SF enjeksiyonları yapıldı ve sonrasında KOH protokolü uygulandı. Deneklerin sakrifikasyonu ile eksize edilen ovaryum dokularında, H&E boyaması ile histomorfolojik değerlendirmeler; ELISA yöntemi ile serum AMH ve TAZ, NAIP, CTGF konsantrasyon tayinleri gerçekleştirildi. VCD enjeksiyonları sonucunda deney gruplarında, ovaryan foliküllerden primordiyal folikül ve primer folikül sayılarında azalma, atretik folikül sayılarında artma, AMH konsantrasyonunda azalma, hippo sinyal yolağı proteinlerinin ekspresyonlarında istatistiksel olarak anlamlı farklar görüldü. Bu çalışmada, sıçan deneysel POI modelinde KOH protokolü öncesi uygulanan intraovaryan PRP enjeksiyonu hippo sinyal yolağının bozulmasında serum fizyolojik enjeksiyonuyla benzer etki gösterdi. Primordiyal foliküllerin aktivasyonunda dokuiçine mekanik uyarılma ile PRP enjeksiyonunun etkilerinin benzer olması ve hipposinyal yolağı proteinleriyle folikül aktivasyon düzeylerindeki uyumsuzluk mekanik uyarılmaya bağlı folikül artışında farklı mekanizmaların araştırılması gerekliliğini gösterdi.
The aim of the thesis study is to comparatively evaluate the effect and mechanism of intraovarian mechanical stimulation and intraovarian PRP injection before thecontrolled ovarian hyperstimulation (COH) protocol on the development of ovarianfollicles through the hippo signaling pathway in cases of premature ovarianinsufficiency (POI), which is one of the causes of infertility.In order to create an experimental POI model and apply intraovarian injectionsand COH protocol, 45 female Sprague-Dawley rats were divided into 7 differentgroups. A POI model was created on the subjects by subcutaneously injecting a singledose of 4-Vinylcyclohexene Diepoxide (VCD, 160 mg/kg/day) for 15 days.Intraovarian following VCD injections; Surgical opening and closing, mechanicalstimulation with an empty needle, PRP and SF injections were performed, and thenthe COH protocol was applied. Histomorphological evaluations with H&E staining inthe ovarian tissues excised after the sacrifice of the subjects; Serum AMH and TAZ, NAIP, CTGF concentration determinations were performed by ELISA method.As a result of VCD injections, a decrease in the number of primordial follicles andprimary follicles from ovarian follicles, an increase in the number of atretic follicles,a decrease in AMH concentration, and statistically significant differences in theexpression of hippo signaling pathway proteins were observed in the experimentalgroups.In this study, intraovarian PRP injection applied before the COH protocol in therat experimental POI model had a similar effect to physiological saline injection indisrupting the hippo signaling pathway. The similarity of the effects of mechanicalstimulation and PRP injection into the tissue on the activation of primordial folliclesand the incompatibility between the Hippo signaling pathway proteins and follicleactivation levels showed the need to investigate different mechanisms in the follicle increase due to mechanical stimulation.
The aim of the thesis study is to comparatively evaluate the effect and mechanism of intraovarian mechanical stimulation and intraovarian PRP injection before thecontrolled ovarian hyperstimulation (COH) protocol on the development of ovarianfollicles through the hippo signaling pathway in cases of premature ovarianinsufficiency (POI), which is one of the causes of infertility.In order to create an experimental POI model and apply intraovarian injectionsand COH protocol, 45 female Sprague-Dawley rats were divided into 7 differentgroups. A POI model was created on the subjects by subcutaneously injecting a singledose of 4-Vinylcyclohexene Diepoxide (VCD, 160 mg/kg/day) for 15 days.Intraovarian following VCD injections; Surgical opening and closing, mechanicalstimulation with an empty needle, PRP and SF injections were performed, and thenthe COH protocol was applied. Histomorphological evaluations with H&E staining inthe ovarian tissues excised after the sacrifice of the subjects; Serum AMH and TAZ, NAIP, CTGF concentration determinations were performed by ELISA method.As a result of VCD injections, a decrease in the number of primordial follicles andprimary follicles from ovarian follicles, an increase in the number of atretic follicles,a decrease in AMH concentration, and statistically significant differences in theexpression of hippo signaling pathway proteins were observed in the experimentalgroups.In this study, intraovarian PRP injection applied before the COH protocol in therat experimental POI model had a similar effect to physiological saline injection indisrupting the hippo signaling pathway. The similarity of the effects of mechanicalstimulation and PRP injection into the tissue on the activation of primordial folliclesand the incompatibility between the Hippo signaling pathway proteins and follicleactivation levels showed the need to investigate different mechanisms in the follicle increase due to mechanical stimulation.
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Keywords
POI, VCD, Mekanik uyarım, PRP, KOH, Hippo sinyal yolağı, Mechanic stimulation, COH, Hippo signaling pathway