UPREGULATION OF NEUROGLOBIN PROMOTES TM3 LEYDIG CELL VIABILITY

Abstract

Neuroglobin (NGB) is a globin family member protein expressed in the nerve system and prevents oxidative stress and apoptosis induced death in neurons. The anti-oxidant role of NGB against oxidative stress and reactive oxygen species (ROS) brings up its promising protective role in other tissues. The overexpression of NGB in testosterone producing Leydig cells might be a solution to hypoxia related male infertility. In the current study, NGB was overexpressed in Leydig cells by using viral transduction methods and the cell proliferation, gene expression and anti-oxidant enzyme levels were analyzed. Upregulation of NGB increased anti-apoptotic Bcl-2, cell proliferation and anti-oxidant enzyme levels and decreased the expression of apoptotic genes BAX, p53 and caspase 3. In addition, NGB transduced cells proliferated and expressed less apoptotic genes after H₂O₂ exposure. In conclusion, NGB might be a target for androgen deficiency related male infertility and could be used in clinics in the future.

Keywords

• Neuroglobin,Leydig cell,oxidative stress,anti-oxidant enzymes,male fertility

UPREGULATION OF NEUROGLOBIN PROMOTES TM3 LEYDIG CELL VIABILITY

Öz

Globin ailesi proteinlerinden bir tanesi olan Neuroglobin (NGB) sinir sisteminde ifade edilmektedir ve oksidatif stres ve apoptoz kaynaklı nöron ölümünü engellemektedir. Oksidatif stres ve reaktif oksijen türlerine karşı gözlemlenen anti-oksidan aktivite, NGB’nin diğer dokulardaki potansiyel koruyucu rolünü gündeme getirmektedir. Testosteron üreten Leydig hücrelerinde NGB’nin fazla ifade edilmesi hipoksi ile ilişkili erkek infertilitesi için bir çözüm olabilecektir. Bu çalışmada, NGB proteinin viral aktarım metodu ile Leydig hücrelerinde aşırı ifade ettirilmiş ve hücre çoğalması, gen anlatımı ve anti-oksidan enzim seviyeleri analiz edilmiştir. Artan NGB, anti-apoptotik Bcl-2, hücre çoğalması ve anti-oksidan enzim seviyelerini arttırmış, BAX, p53 ve kaspaz 3 gibi apoptoz genlerinin ifadesini azaltmıştır. Ek olarak, H₂O₂ uygulaması sonrasında NGB aktarılan hücreler çoğalmış ve apoptotik genlerin ifadesi azalmıştır. Sonuç olarak, NGB androjen eksikliğine bağlı erkek infertilitesi için hedef olabilir ve gelecekte klinikte kullanılabilir.

Anahtar Kelimeler

• Neuroglobin,Leydig cell,oxidative stress,anti-oxidant enzymes,male fertility

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