In Vitro Genotoxicity of Rotenone in Human Lymphocytes

Abstract

Objective: Rotenone is a lipophilic, broad-spectrum, insecticide and piscicide class pesticide that readily crosses cellular membranes. In this study, the effect of Rotenone on DNA of human peripheral blood lymphocytes was evaluated by the comet assay.

Material-Method: In the study, peripheral blood lymphocytes taken from 8 volunteers, 4 male and 4 female, were treated with Rotenone at three different doses as 10, 50 or 100 µM, and for three different time periods of 1, 2 or 4 hours for each dose. Comet assay was applied and tail DNA percentage parameter was chosen as measure of DNA damage and statistically compared with negative and positive control groups.

Results: Rotenone applications showed different results depending on incubation time and dose. Groups incubated with 10 or 50 µM Rotenone for 1 and 2 h caused an increase in DNA damage compared to the negative controls, but this increase was not statistically significant (p>0,05). There was a significant increase in DNA damage in the groups that were incubated with 10, 50 or 100 µM Rotenone for 4 h compared to the negative control group.

Conclusion: When Rotenone exposure is short-term and at low doses, there is an increase in DNA damage, but this increase is not statistically significant. As the dose increases, statistically significant DNA damage also occurs in short-term exposures. Long-term exposure to Rotenone, on the other hand, shows significant DNA damage regardless of dose.

Keywords

• Comet assay
• DNA damage
• Pesticide
• Rotenone

Rotenon’un İnsan Lenfositlerinde İn Vitro Genotoksisitesi

Abstract

Giriş: Rotenon, bir taşıyıcıdan bağımsız olarak hücresel membranları kolayca geçen, lipofilik, geniş spektrumlu insektisit ve pisisit sınıfı bir pestisittir. Bu çalışmada Rotenon’un insan periferik kan lenfositlerinde DNA üzerine olan etkisi comet metodu ile değerlendirilmiştir.

Materyal-Metot: Çalışmada 4 erkek 4 kadın toplam 8 gönüllüden alınan periferik kan lenfositleri Rotenon ile 10, 50 veya 100 µM olmak üzere üç farklı dozda ve her bir doz için 1, 2 veya 4 saat olmak üzere üç farklı sürede muamele edilmiştir. Comet metodu uygulanmış ve kuyruk DNA yüzdesi parametresi DNA hasarının göstergesi olarak negatif ve pozitif kontrol grupları ile istatiksel olarak karşılaştırılmıştır.

Bulgular: Rotenon uygulamaları inkübasyon saatine ve doza bağlı olarak farklı sonuçlar ortaya koymuştur. 10 veya 50 µM Rotenon ile 1 s ve 2 s inkübasyon uygulanan gruplar negatif kontrol gruplarına kıyasla DNA hasarında artışa sebep olmuş ancak bu artış istatistiksel olarak anlamlı bulunmamıştır (p>0,05). 100 µM doz ile 1 ve 2 s inkübasyon uygulanan gruplar, kontrol gruplarına kıyasla DNA hasarında anlamlı artışa sebep olmuştur (p<0,05). 10, 50 veya 100 µM Rotenon ile 4 s inkübasyon uygulanan gruplarda negatif kontrol grubuna kıyasla DNA hasarında anlamlı seviyede artış tespit edilmiştir (p<0,05).

Sonuç: Rotenon maruziyeti kısa süreli ve düşük dozlarda olduğunda DNA hasarında artış olmakla birlikte bu artış anlamlı değildir. Doz yükseldikçe, kısa maruziyet sürelerinde de anlamlı seviyede DNA hasarı oluşmaktadır. Uzun süreli Rotenon maruziyetinde ise doz bağımsız şekilde anlamlı seviyede DNA hasarı görülmektedir.

Keywords

• Comet metodu
• DNA hasarı
• Pestisit
• Rotenon

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