Kafkas Üniversitesi Veteriner Fakültesi Dergisi 2020 , Vol 26 , Issue 5
Protective Effects of Adenovirus-Mediated Overexpression of Heat Shock Protein 70 (HSP70) in Rat Liver Cells Against Oxidative Stress
Jingru GUO1, Huijie HU1, Hongrui LIU1, Jianbin YUAN1, Xu ZHANG1, Hong JI1
1College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, CHINA DOI : 10.9775/kvfd.2020.24079 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 (P<0.01), CAT activity (P<0.01), GSH content (P>0.05), SOD activity (P>0.05), and protein carbonyl content (P<0.01), a lower LDH leakage rate (P<0.01) and GSH-Px activity (P≤0.05), and higher HSP70 mRNA abundance (P<0.01) and HSP70 protein expression level (P<0.01). In summary, Adenovirus-mediated overexpression of HSP70 has certain protective effects on rat liver cells against H2O2-induced oxidative stress. Keywords : HSP70, BRL-3A, Oxidative stress, Adenovirus vector, Cold stress