Hande ŞENOL, Vural KÜÇÜKATAY, Emine Kılıç-TOPRAK, Yasin ÖZDEMİR, Melek Bor-KÜÇÜKATAY, Yalın Tolga YAYLALI

Pulmoner hipertansiyonlu hastalarda comet analizi ile değerlendirilen artmış DNA hasarı

Amaç: Pulmoner hipertansiyonda (PH) görülen patolojik lezyonlar, oksidatif stres ve inflamatuar süreçlerle karakterizedir. Çalışmamızın amacı, PH hastalarında DNA hasarının potansiyel katkısını araştırmaktır. Gereç ve Yöntem: Çalışmamıza 28 tedavi almamış PH hastası (59.93 ± 11.19 yıl) ve 28 yaş-cinsiyet uyumlu (59.86 ± 11.92 yıl) sağlıklı kontrol dahil edilmiştir. Tüm hastalara tanıyı doğrulamak ve hemodinamiyi değerlendirmek için sağ kalp kateterizasyonu uygulanmıştır. Bireylerden alınan venöz kan örneklerinde, DNA hasarı belirlenmesinde tek hücre jel elektroforezi ve flüoresan mikroskopisine dayanan, onarımdan önce DNA hasarını tespit eden kuyruklu yıldız (Comet) analizi kullanılmıştır. Sürekli veriler ortalama ± standart sapma (SD) olarak verilmiş, normallik testi için Shapiro-Wilk testleri kullanılmıştır. Bağımsız gruplar için Mann Whitney U testi, kategorik değişkenler için ki-kare testi kullanılmıştır. İstatistiksel analizlerde SPSS, 24.0 programı kullanılmış ve p≤0.05 değerleri istatistiksel olarak anlamlı kabul edilmiştir. Bulgular: Ortalama pulmoner vasküler direnç 5.64 ± 2.99 Wood Unitesi, ortalama sağ atriyal basınç 9.70 ± 5.38 mmHg, kardiyak indeks 3.12 ± 1.18 l / dak / m2, mixed venöz O2 saturasyonu % 64.77 ± 13.33 olarak saptanmıştır. DNA hasar parametrelerinden, kuyruk uzunluğu (24.02 ± 11.34 ve 16.88 ± 3.55 μm, p = 0.0001), kuyruk momenti (1.93 ± 2.36 ve 0.87 ± 1.03 μm, p = 0.013), kuyruk göçü (10.3 ± 12.24 ve 3.62 ± 2.75, p = 0.03) hastalarda istatistiksel olarak anlamlı düzeyde yüksek saptanmıştır. Sonuç: Bulgularımız, PH hastalarında DNA tamir mekanizmaları devreye girmeden önce DNA hasarının artmış olabileceğini göstermektedir. DNA hasarı PH patofizyolojisine katkıda bulunabilir ve/veya PH'da yeni bir farmakolojik hedef olarak ele alınabilir.

Anahtar Kelimeler:

Pulmoner hipertansiyon, DNA hasarı

Increased DNA damage with comet assay in patients with pulmonary hypertension

Purpose: In pulmonary hypertension (PH), pathologic lesions are characterized by oxidative stress and inflammatory processes. The purpose of our study was to investigate the potential contribution of DNA damage in PH. Materials and methods: The study comprised 28 treatment-naive PH patients (59.93±11.19 years) and 28 age- and sex-matched (59.86±11.92 years) healthy controls. All participants underwent a right-heart catheterization to confirm the diagnosis and to asses hemodynamics. Venous blood was obtained from participants. DNA damage were evaluated using the comet assay which is based on single cell gel electrophoresis and fluorescent microscopy and detects DNA damage prior to repair. Continuous data were reported as mean ± standard deviation (SD). Shapiro–Wilk tests were used for testing normality. Mann Whitney U test was used for independent groups. For categorical variables, Chi-square test was used. SPSS, 24.0 was used for statistical analyses and p value less than 0.05 was considered statistically significant. Results: The average pulmonary vascular resistance was 5.64±2.99 WU, mean right atrial pressure was 9.70±5.38 mmHg, cardiac index was 3.12±1.18 l/min/m2, mixed venous O2 saturation was 64.77±13.33 %. DNA damage parameters such as tail length (24.02±11.34 vs. 16.88±3.55 μm, p=0.0001), tail moment (1.93±2.36 vs. 0.87±1.03 μm, p=0.013), tail migration (10.3±12.24 vs. 3.62±2.75, p=0.03) were significantly higher inpatients with PH. Conslusion: These results suggest that patients with PH could have increased DNA damage before DNA repair mechanisms are at play. DNA damage could be contributing to the pathophysiology and may represent a novel pharmacological target in PH.

Keywords:

pulmonary hypertension, DNA damage,

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