Perfusate composition influences nitric oxide homeostasis in rat juxtamedullary afferent arterioles

Vascular diameters in isolated juxtamedullary nephron preparations perfused with cell-free solutions differ from those perfused with blood. In the present study, the effects of the albumin content of the perfusate on the afferent arteriolar diameter and endothelial nitric oxide were investigated in the isolated juxtamedullary nephron preparation perfused with Krebs-Ringer-bicarbonate buffer containing albumin in different concentrations.

The endothelium was loaded with DAF-FM DA, a nitric oxide-sensitive fluoroprobe. Perfusion was maintained either with 4% (control group), 10 or 20% albumin in the perfusate or with L-NAME (10-4 m) added to the perfusate. Fluorescent images were obtained and stored for evaluation of DAF-FM fluorescence and vascular diameters (in mid-afferent arterioles) immediately before perfusate change and every 15 min thereafter, for a 2-h period.

Increasing the albumin concentration resulted in a decrease in fluorescence. The most rapid decline of fluorescence was obtained following L-NAME administration (relative fluorescence after 2 h: 4% albumin 92.4 +/- 5.3%; 10% albumin 79.5 +/- 4.2%; 20% albumin 66.2 +/- 2.6%; L-NAME 55.4 +/- 3.0%; mean +/- SD, n = 5). A dose-dependent constriction of the afferent arterioles was observed (normalized diameter: 4% albumin 99.8 +/- 3.0%; 10% albumin 80.3 +/- 3.3%; 20% albumin 74.3 +/- 3.2%; L-NAME 70.6 +/- 3.5%).

We propose that albumin interferes with arteriolar nitric oxide homeostasis, probably by scavenging nitric oxide intra-luminally. In this respect, albumin acts similarly to red blood cells in the circulation. The magnitude of the scavenging determines the effectiveness of autoregulation in the perfused preglomerular vessels. The scavenging properties of the perfusing fluid are important in setting operating levels of endothelial nitric oxide.