Effect of endothelial cell-based iNOS gene transfer on cavernosal eNOS expression and mouse erectile responses.
Inducible nitric oxide synthase (iNOS) gene transfer is reported to augment erectile responses in rats, although it is also shown to impair vasorelaxation in cerebral arteries. We investigated the effect of endothelial cell-based iNOS gene transfer on endothelial NOS (eNOS) expression and mouse erectile responses. Human coronary artery endothelial cells (EC) transduced with empty vector (control) or iNOS were grown in culture and transplanted into the corpus cavernosum of severe combined immunodeficient mice. Endothelial NOS expression was compared in control and iNOS-transduced cells grown in the presence or absence of a selective iNOS inhibitor, L-N6- (1-iminoethyl) lysine hydrochloride (L-NIL). At 3-5 days after cell transplantation, we recorded intracorporal pressure (ICP) responses to cavernosal nerve stimulation and measured cavernosal total NO and eNOS protein expression. In this study, EC transduced with iNOS produced significantly more NO than controls but exhibited a twofold downregulation of eNOS protein and mRNA. This effect was reversed by L-NIL. In vivo, the cell-based gene transfer of iNOS led to significantly increased ICP responses, compared to mice transplanted with control ECs. Consistent with the in vitro data, cavernosal lysates had significantly reduced eNOS expression. In conclusion, EC gene transfer of iNOS downregulates EC expression of eNOS by an NOS-dependent mechanism. In the cavernosum of mice transplanted with Inos-transduced EC, nerve-stimulated erectile responses were augmented by the short-term gene transfer. However, our findings suggest that iNOS gene transfer may have deleterious effects on endothelial function if used as a treatment for erectile dysfunction.
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Department of Urology, University of Washington School of Medicine and Harborview Medical Center, Seattle, WA, USA. wessells@u.washington.edu
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Luttrell
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D000818:Animals; D002460:Cell Line; D017690:Cell Transplantation; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D051379:Mice; D016513:Mice, SCID; D052247:Nitric Oxide Synthase Type II / Q000235:genetics* / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics* / Q000378:metabolism*; D009573:Nitrites / Q000378:metabolism; D010410:Penile Erection / Q000502:physiology*; D012190:Retroviridae / Q000235:genetics
C090663:N(6)-(1-iminoethyl)lysine; D009573:Nitrites; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D008239:Lysine