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in
renal medullary interstitial cells
Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226
The present
study was designed to test the hypothesis that hypoxia-inducible
factor-1
(HIF-1)-mediated transcriptional activation contributes
to increased expression of heme oxygenase (HO) genes in renal medullary
interstitial cells (RMICs). By Northern blot analysis, HO-1 mRNA
expression was found to significantly increase in response to reduction
of PO2 in culture medium. However, HO-2 mRNA
was not altered by hypoxia. This hypoxia-induced upregulation of HO-1
mRNA was significantly blocked by HIF-1 inhibition with ferrous
ammonium sulfate. To further determine the role of HIF-1 in the
activation of HO-1, the inducers of HIF-1 were used to address
whether induction of HIF-1 stimulates HO-1 mRNA expression. Both
desferrioxamine and CoCl2 markedly increased HIF-1 mRNA and protein levels and resulted in the upregulation of HO-1 mRNA but
not HO-2. Furthermore, inhibition of HIF-1 degradation by CBZ-LLL,
an inhibitor of ubiquitin-proteasome, significantly increased HIF-1
protein and HO-1 mRNA but not HO-2 in these cells. Using cis-element oligodeoxynucleotide transfection to
specifically decoy HIF-1 and block HIF-1 binding, increased mRNA
expression of HO-1 in response to hypoxia and CoCl2 was
attenuated. In vitro nuclear run-on assays further confirmed that
hypoxia and alterations of HIF-1 mRNA or protein levels
significantly affected the formation of HO-1 mRNA. Taken together, our
results indicate that HO-1, but not HO-2, is transcriptionally
activated by hypoxia through HIF-1-mediated mechanism in RMICs. This
hypoxia-induced transcriptional activation may be one of the important
mechanisms mediating increased expression of HO-1 in the renal medulla.
hypoxia-inducible factor-1; gene transcription; anoxia
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