Herpes Simplex Virus tip 1 inokule edilen Vero hücre kültüründe antioksidan enzim aktiviteleri

Year 2015, Volume: 31 Issue: 2, 122 - 126, 01.06.2015

Sibel Yavru Oguzhan Avci İrmak Dik Kamil Atli

Abstract

Amaç: Bu çalışma Herpes Simplex Virus tip 1 (HSV-1) inokule edilen Vero hücre kültürlerinde antioksidan kapasite/oksidatif stres biomarkırlarını belirlemek amacı ile yapıldı.Gereç ve Yöntem: Vero hücre kültürüne HSV-1 inokule edilerek 21 saat boyunca (1. saate kadar 15 dk’da bir, 6. saate kadar saatte bir, 6-10. saat arası 30 dk’da bir, 10-21. saat arası saatte bir olmak üzere) hücre süpernatantları toplandı. Hücre süpernatantlarındaki süperoksit dismütaz (SOD), katalaz (CAT), glutasyon peroksidaz (GPX) enzimleri, glutasyon (GSH) ve malondialdehit (MDA) miktarı ticari ELISA kitleri ile ölçüldü. HSV-1’in meydana getirdiği sitopatojenik etki (CPE) ise invert mikroskop yardımı ile periyodik olarak değerlendirildi. Bulgular: Araştırmada HSV-1 inokulasyonunu takiben 6. saatte CPE gözlendi. SOD’un en yüksek ve en düşük düzeyi sırasıyla 17. ve 1. saatte belirlenirken, 18. saatte CAT ve 7. saatte GPX seviyeleri en düşük düzeyde tespit edildi. GSH miktarı ilk 30 dk içinde minimum seviyeye ulaştıktan sonra 10. saatte miktarında artış meydana geldi. MDA miktarında 3. saatte ani yükseliş olurken 18. saatte maksimum seviyeye ulaştığı belirlendi. Oksidatif stresin önemli belirteçleri olan SOD ve GSH düzeylerinin HSV-1’in neden olduğu CPE oluşumundan önce azaldığı tespit edildi. Öneri: Lipid peroksidasyonunun HSV-1’in patogenezinde rol alabileceği ve HSV-1 enfeksiyonu tedavisinde antioksidan uygulamalarının faydalı olabileceği ifade edilebilir.

Keywords

HSV-1, SOD, CAT, GPX, GSH

Antioxidant enzyme activities in Vero cell line inoculated with Herpes Simplex Virus type 1

Abstract

Aim: The aim of the present study was to determine of antioxidant capacity/oxidative stress biomarkers in Vero cell line inoculated with Herpes Simplex Virus type 1 (HSV-1).Materials and Methods: Cell supernatants were collected during 21 hours (15 min interval at first hour, 1 hour interval by 6th hour, 30 min interval at 6th -10th hours, 1 hour interval at 6th-21th hour) after HSV-1 inoculations. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) enzymes, glutathione (GSH) and malondialdehyde (MDA) values were analyzed from the test media by commercially available ELISA kits. In addition to this, cytopathogenic effects (CPE) of HSV-1 in cell culture were evaluated periodically by invert microscope.Results: In this research, CPE of HSV-1 was observed at 6th hour post-inoculation. Maximum and minimum levels of SOD were determined at 17th and 1st hours, respectively. Minimum levels of CAT and GPX were determined at 18th and 7th hour, respectively. Minimum level of GSH was determined at first 30 minutes whereas maximum level of GSH was measured at 10th hours. MDA levels suddenly increased at 3rd hours and MDA reached maximum level at 18th hour. The SOD and GSH levels, important markers of oxidative stress, were reduced on HSV-1 before the formation CPE.Conclusion: It is concluded that lipid peroxidation may occur in the pathogenesis of HSV-1 and antioxidant treatment may be useful in the therapy of HSV-1 infection.

Keywords

HSV-1, SOD, CAT, GPX, GSH

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