OKSİRESVERATROLÜN L929 FİBROBLAST HÜCRELERİNDEKİ OKSİDATİF STRES HASARI ÜZERİNE OLUMLU ETKİLERİ

Year 2023, , 543 - 556, 20.05.2023

Fatih Mehmet Ateş , Yasin Bayır

https://doi.org/10.33483/jfpau.1246570

Abstract

Amaç: L929 fibroblast hücrelerinde H2O2 etkisiyle oluşturulan hücre hasarının bertaraf edilmesinde oksiresveratrolün rejeneratif etkisinin değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntem: Oksiresveratrolün total antioksidan seviyesi (TAS), total oksidan seviyesi (TOS), oksidatif stres indeksi (OSİ), toplam fenolik madde seviyeleri farklı derişimlerde ölçülmüştür. Oksiresveratrol fibroblast hücrelerinde IC50 değeri MTT yöntemiyle belirlenmesinin ardından 12.5 - 400 µM konsantrasyonlarında H2O2 ile meydana getirilen oksidatif stres hasarındaki rejeneratif etkisi in-vitro ortamdaki hücre proliferasyonunun ölçümü xCELLigence cihazıyla gerçekleştirilmiştir. Ayrıca hücre hasarındaki yara iyileşme düzeylerini belirlemek amacıyla 3.125 - 25 µM derişimlerinde scratch analizi yapılmıştır.
Sonuç ve Tartışma: Oksiresveratrolün 0.5 mg/ml’de TAS değeri 3 ± 0.3 ve TOS değeri 0.77 ± 0.52; (OSİ) değeri 0.02 ± 0.09 olarak bulunmuştur. Oksiresveratrolün 0.625 mg/ml’den 10 mg/ml’ye farklı dozlarındaki total fenolik madde (TFM) derişimleri düşük konsantrasyondan yüksek konsantrasyona daha yüksek TFM’ye sahip olduğu gözlemlenmiştir. Oksiresveratrolün L929 fibroblast hücreleri IC50 değeri 214.2 μM; H2O2’e maruz bırakılan L929 fibroblast hücrelerindeki hücre hattında oksiresveratrol IC50 değeri 109.7 μM olduğu belirlenmiştir. Oksiresveratrolün 12.5 μM konsantrasyonunun xCELLigence cihazıyla ölçülmesinde en fazla hücre proliferasyonu gözlenmiştir. Scratch analizinde H2O2 ile hasarlanmış hücrelerdeki 12.5 μM oksiresveratrol konsantrasyonunda 24. Saat için %62, 48. Saat için %88’lik bir hücre proliferasyonuyla diğer dozlardan daha fazla iyileşme gözlenmiştir. 12.5 μM konsantrasyonu hem proliferasyon, hem de scratch analizinde en etkili konsantrasyon olduğu belirlenmiştir. Oksiresveratrol; düşük konsantrasyonlarda gösterdiği antioksidan kapasiteyle H2O2 aracılığıyla meydana getirilen hücresel hasarda, L929 fibroblast hücrelerini oksidatif stresten korur, fibroblast canlılığına ve migrasyona bir güç çarpanı etkisi oluşturur. Bu sonuçlar oksiresveratrolün; serbest oksijen radikallerinin oluşturduğu akut ya da kronik hastalıkların engellenmesinde hücre düzeyinde etkili olabileceğini göstermiştir. Hücre düzeyinde gerçekleştirilecek daha kapsamlı çalışmalar, in-vivo çalışmalarla oksidatif stres kaynaklı hastalıkların tedavisinde oksiresveratrolün potansiyel bir molekül olabileceği düşünülebilir.

Keywords

Oksiresveratrol, OSİ, TAS, TFM, TOS

POSITIVE EFFECTS OF OXYRESVERATROL ON OXIDATIVE STRESS DAMAGE IN L929 FIBROBLAST CELLS

Abstract

Objective: It was aimed to determine the regenerative effect of oxyresveratrol in eliminating the cell damage caused by the effect of H2O2 in L929 fibroblast cells.
Material and Method: Total antioxidant capacity (TAC), total oxidant capacity (TOC), oxidative stress index (OSI), total phenolic substance levels of oxyresveratrol were measured at different concentrations. After the IC50 value of oxyresveratrol fibroblast cells was determined by the MTT method, the regenerative effect on oxidative stress damage induced by H2O2 at 12.5 - 400 µM concentrations was performed with the xCELLigence device to measure cell proliferation in-vitro. In addition, scratch analysis was performed at concentrations of 3.125 - 25 µM in order to determine the wound healing levels in cell damage.
Result and Discussion: The TAC value of oxyresveratrol at 0.5 mg/ml was 3 ± 0.3 and the TOC value was 0.77 ± 0.52; (OSI) value was found to be 0.02 ± 0.09. It has been observed that the total phenolic substance (TPS) concentrations of oxiresveratrol at different doses from 0.625 mg/ml to 10 mg/ml have higher TPS from low concentration to high concentration. Oxyresveratrol L929 fibroblast cells IC50 value 214.2 μM; The IC50 value of oxyresveratrol was determined to be 109.7 μM in the cell line of L929 fibroblast cells exposed to H2O2. The highest cell proliferation was observed when measuring the 12.5 μM concentration of oxiresveratrol with the xCELLigence device. Scratch analysis showed a greater improvement than other doses, with a cell proliferation of 62% at the 24th hour and 88% at the 48th hour at a concentration of 12.5 μM oxyresveratrol in H2O2-damaged cells. The 12.5 μM concentration was determined to be the most effective concentration in both proliferation and scratch analysis. Oxyresveratrol; with its antioxidant capacity at low concentrations, L929 protects fibroblast cells from oxidative stress in cellular damage caused by H2O2, and creates a power multiplier effect on fibroblast viability and migration. These results indicate that oxyresveratrol; showed that it can be effective at the cellular level preventing of acute or chronic diseases caused by free oxygen radicals. It can be thought that oxyresveratrol may be a potential molecule in the treatment of oxidative stress-induced diseases with more comprehensive studies to be carried out at the cellular level and in-vivo studies.

Keywords

OSI, oxyresveratrol, TAC, TPS, TOC

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