Datasheet1_Experimental renal transplantation in rats improves cardiac dysfunction caused by chronic kidney disease while LVH persists.docx

BackgroundChronic kidney disease (CKD) causes congestive heart failure (CHF) with systolic dysfunction and left ventricular hypertrophy (LVH), which is a major contributor to increased mortality in CKD patients. It remains unclear whether cardiovascular changes that occur during the course of CKD can be reversed when renal function is restored by transplantation.MethodsTo investigate this, chronic kidney disease was established in F344 rats by subtotal nephrectomy (SNx) for 8 weeks, followed by transplantation of a functional kidney from an isogenic F344 donor. SNx rats without transplantation and sham-operated animals served as controls. Renal function was assessed before and throughout the experiment. In addition, cardiac ultrasound was performed at weeks 0, 8, 12 and 16. At the end of the experiment, intra-arterial blood pressure was measured and kidneys and hearts were histologically and molecularly examined.ResultsEight weeks after SNx, rats developed marked renal dysfunction associated with significant glomerulosclerosis and tubulointerstitial fibrosis, but also an increase in left ventricular mass. After transplantation, renal function normalized but relative heart weight and ventricular mass as assessed by ultrasound scans showed no reduction compared with SNx controls. However, left ventricular wall thickness, fractional shortening and ejection fraction was normalized by renal transplantation. At 8 weeks after kidney transplantation, cardiac expression of BNP and FGF23 was also at levels comparable to healthy controls, whereas these factors were significantly increased in SNx rats. Cardiac fibrosis, as measured by fibronectin mRNA expression, was completely normalized, whereas cardiac fibronectin protein was still slightly but not significantly increased in transplanted animals compared to controls. In addition, the myofibroblast marker collagen 1, as assessed by immunohistochemistry, was significantly increased in SNx rats and also normalized by renal transplantation. Interestingly, CD68+ macrophages were significantly reduced in the hearts of SNx rats and in transplanted animals at slightly higher levels compared to controls.ConclusionRestoration of renal function by kidney transplantation normalized early cardiac changes at most functional and molecular levels, but did not completely reverse LVH. However, further studies are needed to determine whether restoration of renal function can also reverse LVH at a later time point.</p

Table1_Experimental renal transplantation in rats improves cardiac dysfunction caused by chronic kidney disease while LVH persists.xlsx

BackgroundChronic kidney disease (CKD) causes congestive heart failure (CHF) with systolic dysfunction and left ventricular hypertrophy (LVH), which is a major contributor to increased mortality in CKD patients. It remains unclear whether cardiovascular changes that occur during the course of CKD can be reversed when renal function is restored by transplantation.MethodsTo investigate this, chronic kidney disease was established in F344 rats by subtotal nephrectomy (SNx) for 8 weeks, followed by transplantation of a functional kidney from an isogenic F344 donor. SNx rats without transplantation and sham-operated animals served as controls. Renal function was assessed before and throughout the experiment. In addition, cardiac ultrasound was performed at weeks 0, 8, 12 and 16. At the end of the experiment, intra-arterial blood pressure was measured and kidneys and hearts were histologically and molecularly examined.ResultsEight weeks after SNx, rats developed marked renal dysfunction associated with significant glomerulosclerosis and tubulointerstitial fibrosis, but also an increase in left ventricular mass. After transplantation, renal function normalized but relative heart weight and ventricular mass as assessed by ultrasound scans showed no reduction compared with SNx controls. However, left ventricular wall thickness, fractional shortening and ejection fraction was normalized by renal transplantation. At 8 weeks after kidney transplantation, cardiac expression of BNP and FGF23 was also at levels comparable to healthy controls, whereas these factors were significantly increased in SNx rats. Cardiac fibrosis, as measured by fibronectin mRNA expression, was completely normalized, whereas cardiac fibronectin protein was still slightly but not significantly increased in transplanted animals compared to controls. In addition, the myofibroblast marker collagen 1, as assessed by immunohistochemistry, was significantly increased in SNx rats and also normalized by renal transplantation. Interestingly, CD68+ macrophages were significantly reduced in the hearts of SNx rats and in transplanted animals at slightly higher levels compared to controls.ConclusionRestoration of renal function by kidney transplantation normalized early cardiac changes at most functional and molecular levels, but did not completely reverse LVH. However, further studies are needed to determine whether restoration of renal function can also reverse LVH at a later time point.</p

Oxidative DNA Damage in Kidneys and Heart of Hypertensive Mice Is Prevented by Blocking Angiotensin II and Aldosterone Receptors

<div><p>Introduction</p><p>Recently, we could show that angiotensin II, the reactive peptide of the blood pressure-regulating renin-angiotensin-aldosterone-system, causes the formation of reactive oxygen species and DNA damage in kidneys and hearts of hypertensive mice. To further investigate on the one hand the mechanism of DNA damage caused by angiotensin II, and on the other hand possible intervention strategies against end-organ damage, the effects of substances interfering with the renin-angiotensin-aldosterone-system on angiotensin II-induced genomic damage were studied.</p><p>Methods</p><p>In C57BL/6-mice, hypertension was induced by infusion of 600 ng/kg • min angiotensin II. The animals were additionally treated with the angiotensin II type 1 receptor blocker candesartan, the mineralocorticoid receptor blocker eplerenone and the antioxidant tempol. DNA damage and the activation of transcription factors were studied by immunohistochemistry and protein expression analysis.</p><p>Results</p><p>Administration of angiotensin II led to a significant increase of blood pressure, decreased only by candesartan. In kidneys and hearts of angiotensin II-treated animals, significant oxidative stress could be detected (1.5-fold over control). The redox-sensitive transcription factors Nrf2 and NF-κB were activated in the kidney by angiotensin II-treatment (4- and 3-fold over control, respectively) and reduced by all interventions. In kidneys and hearts an increase of DNA damage (3- and 2-fold over control, respectively) and of DNA repair (3-fold over control) was found. These effects were ameliorated by all interventions in both organs. Consistently, candesartan and tempol were more effective than eplerenone.</p><p>Conclusion</p><p>Angiotensin II-induced DNA damage is caused by angiotensin II type 1 receptor-mediated formation of oxidative stress <i>in vivo</i>. The angiotensin II-mediated physiological increase of aldosterone adds to the DNA-damaging effects. Blocking angiotensin II and mineralocorticoid receptors therefore has beneficial effects on end-organ damage independent of blood pressure normalization.</p></div

Additional file 1 of Macrophage subpopulations in pediatric patients with lupus nephritis and other inflammatory diseases affecting the kidney

Additional file 1: Supplemental Table 1. Characteristics of adult LN cohort. Supplemental Table 2. Primary antibodies used for immunofluorescence microscopy. Supplemental Table 3. Secondary antibodies used for immunofluorescence microscopy (IF)

Clinical parameters, parameters of kidney function as well as aldosterone levels of mice after 27 days of angiotensin II-infusion.

<p>* p≤0.05, ** p<0.01, *** p<0.001 vs. Control, ° p≤0.05, °° p<0.01, °°° p<0.001 vs. Ang II treatment.</p><p>Clinical parameters, parameters of kidney function as well as aldosterone levels of mice after 27 days of angiotensin II-infusion.</p

Induction of a marker of renal damage.

<p>Western blot-analysis of the amount of the kidney injury marker KIM-1 (kidney injury molecule) in kidneys of control animals and animals treated with angiotensin II (AngII) with or without co-treatment with candesartan (+Cand), eplerenone (+Eple), or tempol (+Tem). Shown is a representative blot and the quantification of band densities of proteins of all animals.* p≤0.05 vs. Control.</p

Multiple reaction monitoring parameters for the analysis of oxidized bases and internal standard and optimized conditions of the mass spectrometry measurement with the Q-Trap 2000.

<p>Multiple reaction monitoring parameters for the analysis of oxidized bases and internal standard and optimized conditions of the mass spectrometry measurement with the Q-Trap 2000.</p

Histopathological parameters.

<p>GSI: glomerular sclerosis index, MSI: mesangiolysis index, TSI: tubulointerstitial sclerosis index, VSI: vascular sclerosis index, each normalized to the Control values.</p><p>* p≤0.05, ** p<0.01, *** p<0.001 vs. Control, ° p≤0.05, °° p<0.01, °°° p<0.001 vs. Ang II treatment.</p><p>Histopathological parameters.</p

Histopathological changes of kidneys of control animals and animals treated with angiotensin II (AngII) with or without co-treatment with candesartan (+Cand), eplerenone (+Eple), or tempol (+Tem).

<p>A: Representative pictures of PAS-stained tissue, visualizing glomerular damage. B: Representative pictures of HE-stained tissue, visualizing regions of inflammation. C: Representative pictures of Sirius red-stained tissue visualizing regions of fibrosis (red). D: Representative pictures of HE-stained tissue, focussing on changes of the vasculature. Blue filled arrows: examples of mesangiolysis, orange filled arrows: examples of glomerulosclerosis, white filled arrows: examples of infiltrated leukocytes as a marker of inflammation, I–I: device illustrating the thickness of the vessel walls.</p

Primer sequences used for real-time PCR.

<p>NADPH oxidase isoform 1,</p><p>Nox2  =  NADPH oxidase isoform 2, Nox4  =  NADPH oxidase isoform 4.</p><p>Primer sequences used for real-time PCR.</p