Neutrophil-mediated post-ischemic tubular leakage in the rat kidney

Neutrophil-mediated post-ischemic tubular leakage in the rat kidney. Neutropenia was induced in male Sprague-Dawley rats by administration of antineutrophil serum (ANS). A control group received an equal volume of inactive serum. After 45 minutes of unilateral complete renal ischemia the renal blood flow (RBF) was measured by an electromagnetic flow meter. The net filtration force (NFF) in glomerular capillaries, single nephron filtration rate (SNGFR) and frequency of tubular obstructions were estimated by a micropuncture technique. Tubular leakage was measured from the fractional recovery in the normal contralateral kidney of 3H- or 14C-inulin injected into surface proximal and distal tubules of the post-ischemic kidney. Neither ANS nor inactive serum had any influence on inulin clearance (CIn) in the normal kidney. In the post-ischemic kidney, CIn was four times higher in ANS-treated than in control animals. There was no difference in RBF, NFF, SNGFR or the frequency of tubular obstructions between neutrophil-depleted and control animals. The transtubular leakage of inulin injected into proximal tubules was substantially less in the ANS-treated than in the control group (11.3 ± 1.5% vs. 35.1 ± 6.5%; P < 0.01). But distal tubular leakage was equal in the two groups. The control group showed isosthenuria (350 ± 29mOsm · kg-1), while ANS-treated animals produced hyperosmolar urine (555 ± 60mOsm · kg-1; P < 0.05). It is concluded that neutrophil granulocytes mediate post-ischemic tubular leakage, which contributes to the depression in renal clearance parameters and the inability to produce hyperosmolar urine

Comparative tissue transcriptomics reveal prompt inter-organ communication in response to local bacterial kidney infection

<p>Abstract</p> <p>Background</p> <p>Mucosal infections elicit inflammatory responses via regulated signaling pathways. Infection outcome depends strongly on early events occurring immediately when bacteria start interacting with cells in the mucosal membrane. Hitherto reported transcription profiles on host-pathogen interactions are strongly biased towards <it>in vitro </it>studies. To detail the local <it>in vivo </it>genetic response to infection, we here profiled host gene expression in a recent experimental model that assures high spatial and temporal control of uropathogenic <it>Escherichia coli </it>(UPEC) infection within the kidney of a live rat.</p> <p>Results</p> <p>Transcriptional profiling of tissue biopsies from UPEC-infected kidney tissue revealed 59 differentially expressed genes 8 h post-infection. Their relevance for the infection process was supported by a Gene Ontology (GO) analysis. Early differential expression at 3 h and 5 h post-infection was of low statistical significance, which correlated to the low degree of infection. Comparative transcriptomics analysis of the 8 h data set and online available studies of early local infection and inflammation defined a core of 80 genes constituting a "General tissue response to early local bacterial infections". Among these, 25% were annotated as interferon-γ (IFN-γ) regulated. Subsequent experimental analyses confirmed a systemic increase of IFN-γ in rats with an ongoing local kidney infection, correlating to splenic, rather than renal <it>Ifng </it>induction and suggested this inter-organ communication to be mediated by interleukin (IL)-23. The use of comparative transcriptomics allowed expansion of the statistical data handling, whereby relevant data could also be extracted from the 5 h data set. Out of the 31 differentially expressed core genes, some represented specific 5 h responses, illustrating the value of comparative transcriptomics when studying the dynamic nature of gene regulation in response to infections.</p> <p>Conclusion</p> <p>Our hypothesis-free approach identified components of infection-associated multi-cellular tissue responses and demonstrated how a comparative analysis allows retrieval of relevant information from lower-quality data sets. The data further define marked representation of IFN-γ responsive genes and a prompt inter-organ communication as a hallmark of an early local tissue response to infection.</p

Effect of nitric oxide on renal autoregulation during hypothermia in the rat.

Hypothermia-induced reduction of metabolic rate is accompanied by depression of both glomerular perfusion and filtration. The present study investigated whether these changes are linked to changes in renal autoregulation and nitric oxide (NO) signalling. During hypothermia, renal blood flow (RBF) and glomerular filtration rate (GFR) were reduced and urine production was increased, and this was linked with reduced plasma cGMP levels and increased renal vascular resistance. Although stimulation of NO production decreased vascular resistance, blood pressure and urine flow, intravenous infusion of the NO precursor L-arginine or the NO donor sodium nitroprusside did not alter RBF or GFR. In contrast, inhibition of NO synthesis by N(w)-nitro-L-arginine led to a further decline in both parameters. Functional renal autoregulation was apparent at both temperatures. Below the autoregulatory range, RBF in both cases increased in proportion to the perfusion ±pressure, although, the slope of the first ascending limb of the pressure-flow relationship was lower during hypothermia. The main difference was rather that the curves obtained during hypothermia levelled off already at a RBF of 3.9 ± 0.3 mL/min then remained stable throughout the autoregulatory pressure range, compared to 7.6 ± 0.3 mL/min during normothermia. This was found to be due to a threefold increase in, primarily, the afferent arteriolar resistance from 2.6 to 7.5 mmHg min mL(-1). Infusion of sodium nitroprusside did not significantly affect RBF during hypothermia, although a small increase at pressures below the autoregulatory range was observed. In conclusion, cold-induced rise in renal vascular resistance results from afferent arteriolar vasoconstriction by the autoregulatory mechanism, setting RBF and GFR in proportion to the metabolic rate, which cannot be explained by reduced NO production alone

Ischemia causes rapidly progressive nephropathy in the diabetic rat

Ischemia causes rapidly progressive nephropathy in the diabetic rat.We examined the influence of renal ischemia in rats with diabetes mellitus (DM). Male Wistar rats were rendered diabetic by streptozotocin treatment. Two weeks later, 30 minutes of complete ischemia was induced in the left kidney of DM and non-DM animals. Both groups were evaluated functionally and morphologically four or eight weeks post-ischemia. In non-DM animals renal function and morphology showed almost complete recovery. In the DM animals, however, this comparatively short period of ischemia caused a substantial loss of renal function. Four weeks post-ischemia inulin clearance in the DM kidneys rendered ischemic was only 20% of that in the corresponding non-DM kidneys, and after eight weeks the DM kidneys were completely anuric. Extensive inflammation and tubulointerstitial fibrosis were evident in DM kidneys four weeks after ischemia and seemed to increase over time. After eight weeks, tubular atrophy was found in the ischemic DM kidneys, resulting in a substantial loss of kidney mass. We conclude that in diabetic rats renal ischemia causes rapidly progressive kidney damage that in several respects resembles diabetic nephropathy in humans. Since advanced renal lesions similar to those seen in human diabetic nephropathy never develop in the rat solely as a result of DM, the present study may provide an experimental model for further studies on renal failure in diabetes mellitus

Nitric oxide originating from NOS1 controls oxygen utilization and electrolyte transport efficiency in the diabetic kidney

Nitric oxide (NO) is a potent regulator of both vascular tone and cellular oxygen consumption (Qo2). Diabetic kidneys have reduced NO availability and increased Qo2. However, the exact nitric oxide synthase (NOS) isoform regulating Qo2, hemodynamics, and excretory function in the diabetic kidney remains unclear. We therefore investigated the effects of both selective neuronal NOS (NOS1) inhibition and nonselective NOS inhibition. Oxygen utilization, electrolyte transport efficiency [tubular Na+ transport (TNa)/Qo2], renal blood flow (RBF), glomerular filtration rate (GFR), and mean arterial pressure (MAP) were measured in vivo in control and streptozotocin-diabetic rats before and after administration of the selective NOS1 inhibitor S-methyl-l-thiocitrulline (SMTC) or the nonselective NOS inhibitor NG-nitro-l-arginine methyl ester (l-NAME). Diabetic rats had higher baseline Qo2 and GFR than control rats, although RBF was similar in the groups. SMTC and l-NAME increased Qo2 and reduced TNa/Qo2 only in the diabetic animals, whereas both inhibitors increased MAP and reduced RBF in both groups. GFR was reduced by l-NAME, but SMTC had no effect in either group. Carbachol increased RBF and decreased MAP in SMTC-treated rats, whereas it had no effect in l-NAME-treated rats, indicating that SMTC selectively inhibited NOS1. In conclusion, NO regulates RBF and GFR similarly in both control and diabetic rats. However, selective NOS1 inhibition increased Qo2 and reduced TNa/Qo2 in the diabetic rat kidney, indicating a pivotal role of NO produced by NOS1 in maintaining control of Qo2 and tissue oxygenation in these kidneys

Norepinephrine increases calcium sensitivity of mouse afferent arteriole, thereby enhancing angiotensin II–mediated vasoconstriction

Many agents constrict isolated afferent arterioles only at concentrations higher than their physiological levels. Here we determined if norepinephrine, as released by sympathetic nerve activity, could influence the angiotensin II responsiveness of isolated mouse afferent arterioles. Pretreatment of the arterioles for short periods with norepinephrine significantly increased the ability of 10 picomolar angiotensin II to constrict the vessels, an effect inhibited by the alpha receptor blockers prazosin (α-1) or yohimbine (α-2). Although the intracellular calcium transients induced by angiotensin were not different, phosphorylation of the 20kDa myosin light chain was significantly increased in the presence of norepinephrine. Phosphorylation of the p38 mitogen-activated protein kinase was not changed. Phosphorylation of the myosin phosphatase targeting subunit at Thr696, but not at Thr850, was significantly enhanced by, norepinephrine pretreatment, thus increasing the calcium sensitivity of the arteriolar smooth muscle. Our results show that norepinephrine increases afferent arteriolar sensitivity to angiotensin II by means of alpha receptor activation, causing increased calcium sensitivity through phosphorylation of the myosin phosphatase targeting subunit