Esra ARISU NAGHAVI, Bakiye GOKER BAGCA, Senem TEKELİ, Gürkan YİĞİTTÜRK, Burak GÖKÇE, Canberk TOMRUK, Türker ÇAVUŞOĞLU, Çığır AVCI, Cumhur GÜNDÜZ, Yiğit UYANIKGİL

URSODEOKSİKOLİK ASİTİN SİROLİMUS UYGULANAN YAĞ DOKUSU KÖKENLİ MEZENKİMAL KÖK HÜCRELER ÜZERİNE ETKİLERİNİN İNCELENMESİ

Amaç Organ nakli sonrası mezenkimal kök hücrelerin (MKH) immünosupresif ilaçlarla birlikte kullanımı klinik uygulamalarda dikkat çekici hale gelmektedir. Bununla birlikte, ilaçlar MKH'leri olumsuz yönde etkilemektedir. Antioksidan bir molekül olan ursodeoksikolik asit (UDKA) bu etkileri tersine çevirebilecektir. Bu çalışmanın amacı, sirolimus ve UDKA'nın bireysel ve kombinasyon olarak uygulanmasının insan yağ dokusu kaynaklı MKH'ler (YDKMKH) üzerindeki etkilerinin incelenmesidir. Gereç ve Yöntem Etken maddelerin sitotoksik etkileri zamana ve doza bağlı WST-1 testi ile değerlendirildi. Kombinasyon etkileri, izobologram analizi kullanılarak belirlendi. Apoptoz ve hücre döngüsünün değerlendirilmesi için Muse hücre analizörü kullanıldı. Oksidatif stress belirteçlerinin değişimi biyokimyasal yöntemle ölçüldü. Bulgular Sirolimusun IC50 dozu 48. saatte 18.58μM olarak belirlendi. UDKA uygulanan doz aralığında sitotoksik etki belirlenmediği için apoptoz, hücre döngüsü ve oksidatif stres indikatör analizlerine 100 μM güvenli doz ile devam edildi. Sirolimusun, apoptozu teşvik ettiği ve hücre proliferasyonunu inhibe ettiği belirlendi. UDKA'nın antioksidan özelliği ile sirolimusun YDKMKH'ler üzerindeki apoptotik ve antiproliferatif etkilerini azalttığı belirlendi. Sonuç Organ ve doku transplantasyonu sonrası immünosupresif tedavi ile kombinasyon halinde UDKA tedavisinin YDKMKH'ler üzerinde olumlu etkileri olabilecektir.

Anahtar Kelimeler:

sirolimus, ursodeoksikolik asit, oksidatif stres, insan yağ dokusu kaynaklı mezenkimal kök hücreler

INVESTIGATION OF URSODEOXYCHOLIC ACID EFFECTS ON SIROLIMUS TREATED ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS

Objective The usage of mesenchymal stem cells (MSC) with immunosuppressive drugs after organ transplantation is becoming remarkable in clinical applications. However, the drugs negatively affect MSCs. Ursodeoxycholic acid (UDCA), which is an antioxidant molecule, may reverse these effects. The study aims that to determine the effects of sirolimus and UDCA on human adipose tissue-derived MSCs (ADMSCs) individually and in combination. Material and Method The cytotoxicity of the agents was evaluated by WST-1 test in time and dose-dependent manner. The combinational effects were determined using isobologram analysis. Muse cell analyzer was used for the evaluation of apoptosis and cell cycle. Oxidative stress markers were measured by biochemical methods. Results IC50 dose of sirolimus was determined as 18.58μM in the 48th hour. Because no cytotoxic effect was observed at the studied doses of UDCA, the apoptosis, cell cycle, and oxidative stress indicator analyses were continued with a safe dose of 100 μM. Sirolimus promoted apoptosis and inhibited cell proliferation. It was determined that UDCA reduced the apoptotic and anti-proliferative effects of sirolimus on ADMSCs with its anti-oxidant property. Conclusion The UDCA treatment in combination with immunosuppressive therapy after organ and tissue transplantation may have positive effects on ADMSCs.

Keywords:

Sirolimus, Ursodeoxycholic acid, Oxidative stress, Human adipose tissue-derived mesenchymal stem cells,

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