Proximal tubular fluid angiotensin II levels in angiotensin II-induced hypertensive rats

BACKGROUND: It has been shown that infusions of low-dose angiotensin II (Ang II) for 2 weeks lead to impaired pressure natriuresis and autoregulatory capability. Although intrarenal renin content and renin mRNA levels are markedly reduced, whole-kidney Ang II content has been shown to be increased. However, the intrarenal distribution of the increased intrarenal Ang II has not been established. OBJECTIVE: To determine the concentrations of Ang II in the proximal tubule fluid achieved in hypertensive rats (n = 16) infused with Ang II, previously prepared by infusion with Ang II at 60 ng/min via osmotic minipump for 13 days. METHODS: Rats were anesthetized with pentobarbital sodium and prepared for micropuncture, and then several free-flow proximal tubular fluid collections were obtained and pooled for each rat. At the end of each experiment, a blood sample was collected and the micropunctured kidney was excised and homogenized in chilled methanol. All samples were extracted immediately after collection and stored at 20 degrees C until the day of Ang II radioimmunoassay. RESULTS: Mean arterial blood pressure averaged 179 +/- 3 mmHg, renal plasma flow was 1.89 +/- 0.15 ml/min per g, and glomerular filtration rate averaged 0.58 +/- 0.04 ml/min per g. The Ang II concentration in proximal tubular fluid averaged 4.5 +/- 1.1 pmol/ml, a value substantially greater than the Ang II concentrations in plasma (0.17 +/- 0.03 pmol/ml), urine (0.06 +/- 0.01 pmol/ml), or total kidney tissue (0.40 +/- 0.10 pmol/g). Plasma renin activity (1.0 +/- 0.21 ng Ang I/ml per h) was markedly suppressed, as observed previously. CONCLUSIONS These findings indicate that Ang II concentrations in proximal tubular fluid collected from kidneys of anesthetized hypertensive rats infused with Ang II are in the nanomolar range, similar to those observed in normotensive rats. The inappropriate maintenance of nanomolar concentrations of Ang II in proximal tubular fluid of Ang II-infused hypertensive rats, even at markedly increased arterial pressures, may contribute to the impaired pressure natriuresis capability previously reported and, thereby, to the development and maintenance of hypertension in this model.