Objective    To establish a novel stain of renal cell (HK-2) overexpressing Bcl-2 protein and investigate whether the overexpression of Bcl-2 protein has the anti-apoptosis effect on renal tubule epithelial cells (RTEC) in rats with acute renal failure (ARF).     Methods    Bcl-2 retrovirus

in the supernatants of packaging cell PA317 was used to infect the human kidney epithelial cell line HK-2. The G418 resistant colonies were selected

and PCR-verified positive colony was named as HK-2

Bcl-2

. Hybridization in situ was employed to determine the Bcl-2 mRNA levels in this cell strain. ARF rats were

induced by intramuscular injection of glycerol at a large dose of 1.5 ml per 100 g body weight

or a small dose of 0.75 ml. Then 1.8×10

5

CFU Bcl-2 retrovirus in the supernatant of packaging cell PA317 was infused to the rats of virus intervention group via bilateral renal arteries. Thus

there were 5 groups

group A (simple glycerol injection)

group B (large dose of glycerol and virus intervention)

group C (small dose of glycerol and virus intervention)

group D (simple virus intervention)

and group E (normal saline injection). Their kidney samples were taken 24 and 96 h

7

10

14

and 21 d after intervention. Immunohistochemical assay was used to detect the expressions of Bcl-2 and PCNA proteins

and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was employed for apoptotic cells.     Results    Hybridization in situ showed that the cells in G418 resistant HK-2 colonies overexpressed Bcl-2

indicating the establishment of a new stain of HK-2 overexpressing Bcl-2 protein. In ARF rats

Bcl-2 retrovirus infected renal tubular epithelium

up-regulated the expression of Bcl-2 protein

and had anti-apoptosis effect on renal tubular epithelial cells.     Conclusion    A stable renal epithelial cell line HK-2

Bcl-2

that overexpresses Bcl-2 mRNA is established successfully. The reconstructed Bcl-2 retrovirus can in vivo obviously inhibit the apoptosis of renal tubular epithelial cells.