Formik asidin insan lenfositleri üzerindeki in vitro genotoksik ve sitotoksik etkisi

Öz

Amaç: Formik asit, bitkiler, arılar, toprak, araçlar, gübre yanması ve fotokimyasal reaksiyonlar gibi çeşitli kaynaklarca üretilen ve çevrede yaygın olarak bulunan kimyasal bir bileşimdir. Mevcut çalışma, insan lenfositlerinde sitokinez-blok mikronükleus tayini ve kromozomal aberasyon analizi gibi sitogenetik testler kullanarak formik asidin sitotoksik ve genotoksik etkilerinin in vitro analizine odaklanmıştır. Yöntem: Bu çalışma 18-22 yaş arası 10’u erkek, 6’sı kadın sağlıklı, sigara kullanmayan yetişkinlerinden kan örnekleri kullanılarak gerçekleştirilmiştir. Lenfosit kültürüne kromozomal aberasyon CA testi için formik asidin farklı konsantrasyonları 0,07, 0,1, 0,2, 0,3, 0,4, 0,5, 0,8 mM eklendi. Pozitif kontrol olarak mitomycin-C 0,3 µg/ml kullanıldı. Sitokinez-blok mikronükleus tayini CBMN için, insan periferik kan lenfositleri 20, 40, 60, 80 mM derişimde formik asitle 48 saat işleme tabi tutuldu. Lenfosit kültürüne, pozitif kontrol olarak mitomycin-C 0,50 µg/ml eklendi. Bu araştırma, sitokinezblok mikronükleus tayinini CBMN ve kromozomal aberasyon CA analizini kullanarak in vitro insan periferal lenfositlerinde formik asidin sitotoksik ve genotoksik etkilerini değerlendirmek için yapılmıştır. Bulgular: Pozitif kontrol olarak kullanılan mitomisin C MMC, 0,5 µg/ml ile negatif kontrol karşılaştırıldığında formik asitin tüm dozları mikronükleus frekansını belirgin bir düzeyde arttırdığı gözlenmiştir. Pozitif kontrol olarak kullanılan mitomisin C MMC, 0,3 µg/ml ’nın negatif kontolle karşılaştırıldığında formik asidin 24 saatte tüm uygulama derişimlerinde, kromozomal aberasyonların sıklığını belirgin bir şekilde arttırdığı gözlenmiştir. Mikronükleus sıklığı ve kromozomal aberasyonlar, doza bağımlı olarak artmıştır. Sonuçlar formik asit konsantrasyonu, mikronükleus frekansı r=0,92 , nekrotik hücrelerin sayısı r=0,95 ve apoptotik hücreler r=0,91 arasında anlamlı korelasyonların var olduğunu göstermektedir. Sonuç: Verilerimiz, in vitro olarak formik asit konsantrasyonu ve mikronükleus frekansı, apoptotik hücreler ile nekrotik hücreler gibi kromozomal aberasyonlar arasında anlamlı bir korelasyon olduğu kanıtını sağlamıştır

Anahtar Kelimeler

• Formik asit, insan lenfosit kültürü, kromozomal aberasyon, mikronükleus, sitotoksisite

Genotoxic and cytotoxic effects of formic acid on human lymphocytes in vitro

Abstract

Objective: Formic acid is an ubiquitous chemical constituent in the environment, being produced by sources as diverse as vegetation, ants, soil, vehicles, biomass burning, and photochemical reactions. The present work is focused on in vitro analysis of cytotoxic and genotoxic effects of formic acid, using cytogenetic tests such as the cytokinesis-block micronucleus assay CBMN and chromosomal aberration analysis, in human lymphocytes. Method: This study was carried out using blood samples from healthy, non-smoking adults aged 18–22 years, of whom 10 were male and 6 were famale. Different concentrations 0.07, 0.1, 0.2, 0.3, 0.4, 0.5, 0.8 mM of formic acid was added to the lymphocyte culture test for chromosomal aberration CA analysis. Mitomycin-C 0.3 mg/ml was used as the positive control. Human peripheral blood lymphocyte cells were treated with 20, 40, 60, 80 mM concentrations of formic acid for 48 h. for the CBMN test. Mitomycin-C 0.5 mg/ml was added to the Lymphocyte culture as a positive control. The present research was carried out to assess the cytotoxic and genotoxic effects of formic acid on human peripheral lymphocytes in vitro using the cytokinesis-block micronucleus assay CBMN , as well as chromosomal aberration CA analysis. Results: A significant increase was observed for induction of micronucleus frequency in all treatments of formic acid concentrations for 48 h. comparing with the negative control and mitomycin C MMC, 0.5 µg/ ml which was used as positive control. When compared with negative control and with mitomycin C MMC, 0.3 µg/ml which is used as positive control, it is observed that formic acid is rising the frequency of chromosomal aberration significantly at all appliance concantrations in 24 h. The frequency of micronuclei and chromosomal aberrations increased in a dose dependent manner. The results showed that there were significant correlations between formic acid concentration and micronuclei frequency r= 0.92 , numbers of necrotic cells r= 0.95 , and apoptotic cells r= 0.91 . Conclusion: Our data provided evidence that there is a significant correlation between the concentration of formic acid and the following chromosomal aberrations: frequency of micronuclei, apoptotic cells, and necrotic cells in vitro.

Keywords

• Formic acid, human lymphocyte culture, chromosomal aberration, micronucleus, cytotoxicity

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