Pulmoner hipertansiyonlu hastalarda comet analizi ile değerlendirilen artmış DNA hasarı

Year 2019, , 415 - 421, 30.09.2019

Emine Kılıç-toprak Yalın Tolga Yaylalı Yasin Özdemir Vural Küçükatay , Hande Şenol , Melek Bor-küçükatay

https://doi.org/10.31362/patd.557153

Abstract

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.

Keywords

Pulmoner hipertansiyon, DNA hasarı

Increased DNA damage with comet assay in patients with pulmonary hypertension

Abstract

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 in

patients
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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