[Protection of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by recombinant adenovirus containing hCGPx gene].
OBJECTIVE
To study the protective effect of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by transfection of recombinant adenovirus-mediated human cytosolic glutathione peroxidase(hCGPx) gene.


METHODS
Recombinant pGEM-T vector containing hCGPx cDNA and pACCMV-pLpA adenovirus shuttle plasmid was constructed. Then the shuttle plasmid pACCMV-hCGPx and pJM17 were co-transfected into 293 cells and recombinant adenovirus AdCMV-hCGPx was obtained. Cultured HK-2 cells were transfected with AdCMV-hCGPx or vacant recombinant adenovirus (control). The expression ratio of transfected hCGPx gene were studied. Cell viability, the percentage of apoptosis and death were evaluated after hypoxia-reoxygenation damage.


RESULTS
The expression ratio of hCGPx gene was higher in the AdCMV-hCGPx transfected cells than that in the control group (P<0.01). After hypoxia-reoxygenation damage, the viability of hCGPx gene transfected cells was significantly higher than that of control and the percentage of apoptosis and death of hCGPx transfected cells was significantly lower than that of control.


CONCLUSIONS
The transfection of hCGPx mediated by recombinant adenovirus could protect renal tubular epithelial cells from hypoxia-reoxygenation damage in vitro.
Xiang
He-li
HL
Department of Kidney Transplantation, First Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710061, China. hlxiang@163.com
Xue
Wu-jun
WJ
Hou
Jun
J
Tian
Pu-xun
PX
Teng
Yan
Y
Pan
Xiao-ming
XM
Ding
Xiao-ming
XM
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis; D003600:Cytosol / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D000860:Hypoxia / Q000378:metabolism*; D007684:Kidney Tubules / Q000473:pathology*; D010100:Oxygen / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D004274:DNA, Recombinant; D005979:Glutathione Peroxidase; D004262:DNA Restriction Enzymes; D010100:Oxygen