Melih AKYOL, Mustafa TOSUN, Sedat ÖZÇELİK, Sibel BERKSOY HAYTA

Changes emerging with calcinerin inhibition in Caenorhabditis elegans individuals as an aging model

Amaç: Kalsinörin inhibisyonunun Caenorhabditis elegans (C. elegans) bireylerinde yaşam süresini uzattığı bilinmektedir. C. elegans genlerinin önemli bir kısmı insan genlerine büyük benzerlik göstermektedir. Bu çalışmada kalsinörin inhibisyonunun C. elegans bireylerindeki yaşamsal fonksiyonlar üzerine olan etkilerinin değerlendirilmesi amaçlandı. Yöntem: Çalışma için bir kalsinörin inhibitörü olan siklosporin A'nın 1 ?M (1.doz), 0.1 ?M (2.doz), 0.01 ?M (3.doz) ve 0.001 ?M (4.doz) dozunda çözeltileri hazırlanarak her bir çözeltiden 1'er ml, içlerinde 10' ar ml Nematod Growth Media (NGM) bulunduran petrilere eklenmiştir. Kontrol grubu siklosporin eklenmemiş besiyerinde beslenmiştir. Senkronizasyonu yapılmış C. elegans'lardan siklosporinin belirtilen dozları ile hazırlanmış her bir petriye 20' şer adet aktarılmıştır. 6X-50X arasında büyütme sağlayan alttan ışıklandırmalı stereo mikroskop ile C. elegans'lar izlenmiştir. Bütün petrilerdeki C. elegans'lar ölene kadar her gün aynı saatte canlı hayvanlar sayılmış ve kontrol grubuyla karşılaştırılmıştır. Bulgular: Çalışma sonucunda siklosporine maruz bırakılan C. elegans'ların kontrol grubuna göre yaşam sürelerinde doz artışına bağlı istatistiksel olarak anlamlı bir artış olduğu tespit edilmiştir (p< 0,05). Siklosporine maruz bırakılan C. elegans'ların fiziksel gelişimlerinin doz artışına bağlı olarak anlamlı derecede azaldığı saptanmıştır (p< 0,05). Siklosporin dozundaki artış farinks pompalama sayısında istatistiksel olarak anlamlı bir değişikliğe neden olmamıştır (p> 0,05). Sonuç: Kalsinörin inhibisyonu, C. elegans'ların gelişimsel sürecini yavaşlatır ve yaşam süresini uzatır. Kalsinörin inhibitörleri, benzer mekanizmalarla insanlarda deri yaşlanması üzerine olumlu etki yapabilir. Bu nedenle kalsinörin inhibisyonunun, deri yaşlanması üzerine etkinliğini ve etki mekanizmasını açıklayacak ileri çalışmalar gereklidir.

Bir yaşlanma modeli olarak Caenorhabditis elegans bireylerinde kalsinörin inhibisyonu ile ortaya çıkan değişiklikler

Objective: It is known that calcineurin inhibition prolongs life in individuals with Caenorhabditis elegans (C. elegans). An important part of C. elegans genes shows great similarity to human genes. This study aims to evaluate the effects of a calcineurin inhibitor on vital functions in C. elegans individuals. Method: In this study, solutions of cyclosporine A which is a calcineurin inhibitör 1 ?M (1.dose), 0.1 ?M (2.dose), 0.01 ?M (3.dose) and 0.001 ?M (4.dose) were prepared and 1 ml from each solution was taken and added to petri dishes, containing 10 ml Nematode Growth Media (NGM). The control group was fed the medium which was not added cyclosporine. The indicated doses of cyclosporine whose synchronization has been completed were transferred to petri dishes in order that each would have 20 pieces. C. elegans were observed using bottom light stereo microscop that provided 6X-50X magnification. Live animals at the same time every day were counted and compared with the control group until all C. elegans in all petri dishes died. Results: The results have showed that there was a statistically significant increase in life span periods of C. elegans exposed to cyclosporin depending on dose increase (p< 0,05). It has been found out that physical development of C. elegans exposed to increased doses of cyclosporin decreased significantly (p< 0,05). The increase in doses of cyclosporin have not caused statitically significant change in the number of pharynx pumping (p> 0,05). Conclusions: Calcineurin inhibition slows down the developmental process in C. elegans and prolongs survival, and may produce positive effects on aging.

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