Sigara içen ve içmeyen sağlıklı erkek bireylerde agız içi mukoza epitel hücrelerinin mikronükleus (MN) ve nükleolus organizer bölgeleri (NOR'lar) üzerinde ışık mikroskobik çalışmalar
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2009
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Selçuk Üniversitesi Sağık Bilimleri Enstitüsü
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Abstract
Bu çalışmada, günde bir paket sigara içen (20 adet/gün) sağlıklı gönüllü erkek bireylerden, abeslangla alınan dil ve yanak mukoza kazıntıları materyal olarak kullanıldı. Her birinde 30'ar kişi bulunan 35?45, 46?55, 56 ve üzeri olmak üzere üç yaş grubu oluşturuldu. Havada, oda sıcaklığında (20ºC) kurutulan frotilerden AgNOR boyamasında kullanılacak olanlar metanol-asetik asit karışımında (3:1), MN boyaması uygulanacaklar ise glutaraldehid-aseton tespit sıvısında 5 dakika tespit edildi. MN boyaması, May Grünwald-Giemsa boyasının modifikasyonuyla geliştirilen Wolf ve Luepke'nin (1997) uyguladığı metoda göre gerçekleştirildi. AgNOR'lar Aydın (2004) tarafından bildirilen gümüşleme boyama yöntemiyle boyandı. Modifiye May Grünwald-Giemsayla boyanan preparatlarda toplam 1000 adet epitel hücresindeki MN sayıları belirlendi ve sonuçlar, 1000 hücre başına düşen MN olarak ifade edildi. AgNOR boyayanan preparatlardaysa, dijital görüntü analiz programıyla her hücrenin çekirdek alanı, AgNOR sayısı ve AgNOR alanı belirlendi. Elde edilen verilerden, AgNOR alanının çekirdek alanı içindeki yüzdesi (nisbi, % AgNOR alanı) hesaplandı. Bu çalışma sonuçları, sigara içiminin, dil ve yanak mukoza epitel hücrelerinde genotoksik etkiye sahip olduğu hakkında daha önce ileri sürülen bulguları desteklemektedir. Sigara içenlerde, hem dil ve hem de yanak mukoza epitel hücrelerinin MN frekansında önemli artış gözlenmektedir. Nükleus büyüklüğü ve AgNOR parametrelerinde de sigara içenlerde bazı değişiklikler meydana gelmekle birlikte, dil ve yanak mukoza epitel hücreleri arasında farklar vardır.
In this study, lingual and buccal mucosal scrapings of healthy non smoking and smoker (20 cigarettes/day) male volunteers from three age groups (35?45 years, 46?55 years, 56 and elder), each having 30 people, were used as materials. From each scrapings, smears were prepared, air dried at room temperature (20ºC) and fixed in methanol-acetic acid (3:1, v/v) for AgNOR staining, some of them fixed in glutaraldehyde-acetone for MN staining, for 5 minutes. MNs were stained by a modified May Grünwald-Giemsa stain (Wolf and Luepke 1997). AgNORs were stained by means of a silver impregnation method reported by Aydın (2004). In modified May Grünwald-Giemsa stained specimens, MNs were carefully counted under light microscope in 1000 epithelial cells for each sample, and the results were expressed as MN frequency, MN number/1000 cells. Digital images of AgNOR stained specimens were analysed by an image analysing software, and nuclear area, AgNOR numbers and AgNOR area of 200 cell for each sample were determined. Relative AgNOR area (% area of total AgNOR area in each nuclear area) were calculated from the data. The results were in accordance with previous findings showing that smoking had genotoxic effects on the epithelial cells of both lingual and buccal mucosae. Smokers had increased MN frequencies in lingual and buccal epitheial cells. Nucleus size and AgNOR parameters were also affected by cigarette smoking, although there were some differences between the lingual and buccal epithelial cells.
In this study, lingual and buccal mucosal scrapings of healthy non smoking and smoker (20 cigarettes/day) male volunteers from three age groups (35?45 years, 46?55 years, 56 and elder), each having 30 people, were used as materials. From each scrapings, smears were prepared, air dried at room temperature (20ºC) and fixed in methanol-acetic acid (3:1, v/v) for AgNOR staining, some of them fixed in glutaraldehyde-acetone for MN staining, for 5 minutes. MNs were stained by a modified May Grünwald-Giemsa stain (Wolf and Luepke 1997). AgNORs were stained by means of a silver impregnation method reported by Aydın (2004). In modified May Grünwald-Giemsa stained specimens, MNs were carefully counted under light microscope in 1000 epithelial cells for each sample, and the results were expressed as MN frequency, MN number/1000 cells. Digital images of AgNOR stained specimens were analysed by an image analysing software, and nuclear area, AgNOR numbers and AgNOR area of 200 cell for each sample were determined. Relative AgNOR area (% area of total AgNOR area in each nuclear area) were calculated from the data. The results were in accordance with previous findings showing that smoking had genotoxic effects on the epithelial cells of both lingual and buccal mucosae. Smokers had increased MN frequencies in lingual and buccal epitheial cells. Nucleus size and AgNOR parameters were also affected by cigarette smoking, although there were some differences between the lingual and buccal epithelial cells.
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Sigara içme, Smoking, Mikronükleus, Micronucleus
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Doğan, F. (2009). Sigara içen ve içmeyen sağlıklı erkek bireylerde agız içi mukoza epitel hücrelerinin mikronükleus (MN) ve nükleolus organizer bölgeleri (NOR'lar) üzerinde ışık mikroskobik çalışmalar. Selçuk Üniversitesi, Yayımlanmış yüksek lisans tezi, Konya.
