Protective Effects of Adenovirus-Mediated Overexpression of Heat Shock Protein 70 (HSP70) in Rat Liver Cells Against Oxidative Stress

This study was aimed to study the protective effects of Adenovirus-mediated overexpression of heat shock protein 70 (HSP70) in rat liver cells against oxidative stress. Cultured Buffalo Rat Liver cells (BRL-3A) cells were divided into six groups. Groups SH and CH were transfected with Ad-CMV-HSP70, groups SN and CN were transfected with Ad-CMV-Null, and groups SC and CC were treated under the same conditions without adenovirus for 48 h. Groups SH, SN and SC were treated with 500 μM H2O2 for 3 h, and groups CH, CN and CC were treated under the same conditions without H2O2. The following parameters were determined for cells of each group: proliferation rate, lactic dehydrogenase (LDH) leakage rate, catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) enzyme activities, and glutathione (GSH) and protein carbonyl contents. In addition, the HSP70 mRNA abundance and HSP70 protein expression levels were examined using quantitative real-time PCR (qRT-PCR) and western blotting, respectively. The result shows that compared with cells of group SC, cells of group SH had a higher proliferation rate (P0.05), and protein carbonyl content (P

Sıçan Karaciğer Hücrelerinde Isı Şok Protein 70 (HSP70)’in Adenovirüs Aracılı Aşırı Ekspresyonunun Oksidatif Strese Karşı Koruyucu Etkileri

Bu çalışmada, sıçan karaciğer hücrelerinde ısı şok protein 70 (HSP70)’in Adenovirüs aracılı aşırı ekspresyonunun oksidatif strese karşı koruyucu etkilerinin araştırılması amaçlamıştır. Kültürlenmiş Buffalo Rat Karaciğer hücreleri (BRL-3A) altı gruba ayrıldı. SH ve CH grupları Ad-CMV-HSP70 ile, SN ve CN grupları Ad-CMV-Null ile transfekte edildi. SC ve CC grupları ise 48 saat boyunca aynı koşullar altında adenovirüs olmadan tedavi edildi. SH, SN ve SC gruplarına 3 saat boyunca 500 μM H2O2 uygulandı ve CH, CN ve CC grupları H2O2 olmadan aynı koşullara maruz bırakıldı. Her bir gruptaki hücreler için aşağıdaki parametreler belirlendi: proliferasyon hızı, laktik dehidrogenaz (LDH) sızıntı oranı, katalaz (CAT), glutatyon peroksidaz (GSH-Px) ve süperoksit dismutaz (SOD) enzim aktiviteleri, glutatyon (GSH) ve protein karbonil içeriği. Ek olarak, HSP70 mRNA yoğunluğu ve HSP70 protein ekspresyon seviyeleri, sırasıyla kantitatif gerçek zamanlı PCR (qRT-PCR) ve western blot yöntemi kullanılarak incelendi. Sonuçlar, SC grubundaki hücrelerle karşılaştırıldığında, SH grubundaki hücrelerin daha yüksek proliferasyon oranına (P0.05), ve protein karbonil içeriğine (P

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