[Recombinant adenovirus-mediated human cytosolic glutathione peroxidase gene transfection protects vascular endothelial cells from oxidative damage].
OBJECTIVE
To study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage.


METHODS
pGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2).


RESULTS
The expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage.


CONCLUSIONS
hCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.
Xiang
He-li
HL
Department of Kidney Transplantation, First Affiliated Hospital, Medical College of 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
Feng
Xin-shun
XS
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003600:Cytosol / Q000201:enzymology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005434:Flow Cytometry; D005822:Genetic Vectors; D005979:Glutathione Peroxidase / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D018384:Oxidative Stress; D010957:Plasmids / Q000235:genetics; D013997:Time Factors; D014162:Transfection
D006861:Hydrogen Peroxide; D005979:Glutathione Peroxidase