[Recombinant heat shock protein 70 induced by adenovirus protects neurons and glial cells against hypoxia/reoxygenation injury].
OBJECTIVE
To investigate the protective effect on neurons and glial cells against hypoxia/ reoxygenation injury with recombinant heat shock protein 70 (HSP70) induced by adenovirus.


METHODS
Neurons and glial cells in culture were divided into four groups: three groups were treated with recombinant adenovirus (vAd-HSP70) transfected human HSP70 gene at 24, 48 and 72 hours respectively, and vAd-GFP transfected cell served as control. Cells in different groups were subjected to hypoxia/reoxygenation, then the cell viability was analyzed by methyl thiazolyl tetrazolium (MTT) method, lactate dehydrogenase (LDH) viability was evaluated with LDH staining kit, and cytochrome C (Cyt C) in mitochondria and cytoplasm were assessed by Western blotting.


RESULTS
The expression of human HSP70 gene was detected in the vAd-HSP70 transfection group. After hypoxia/reoxygenation treatment, the cell viability in transfected groups was higher than that of control group (all P<0.05), the LDH viability of vAd-HSP70 transfected groups at different time points was 1,480+/-121, 1,023+/-106, and (1,132+/-197) U/L respectively, and they were significantly lower than control group [(1,976+/-190) U/L, all P<0.01]. In transfected groups, the content of Cyt C in mitochondria (0.986+/-0.012, 1.028+/-0.007, 1.014+/-0.008) was significantly higher than control group (0.970+/-0.003, P<0.05 or P<0.01). In contrast, the content of Cyt C in cytoplasm (0.987+/-0.008, 0.960+/-0.005, 0.964+/-0.003) was lower than that of control group (1.011+/-0.005, all P<0.01). The protective effect was especially obvious when the cells were transfected by vAd-HSP70 at 48 hours (all P<0.01).


CONCLUSIONS
The expression of human HSP70 mediated by recombinant adenovirus may protect neurons and glial cells against hypoxia/reoxygenation in vitro.
Yang
Fang-fang
FF
Intensive Care Unit, The Affiliated Qingdao Municipal Hospital of Qingdao University Medical College, Qingdao 266011, Shandong, China. amy820211@yahoo.cn
Xu
Xiao-na
XN
Hu
Dan
D
Shi
Fei
F
Qu
Yan
Y
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015687:Cell Hypoxia; D002470:Cell Survival; D002478:Cells, Cultured; D005822:Genetic Vectors; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000627:therapeutic use*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000627:therapeutic use; D014162:Transfection
D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins; D007770:L-Lactate Dehydrogenase