Urinary MCP-1 Research Articles

MCP-1 in urine as biomarker of renal lupus in absence of cytokines, interferon-γ and growth factors

ObjectiveTo characterize 17 immunological markers in the Urine of patients with SLE. IntroductionLupus nephritis is an inflammatory disease affecting the renal parenchyma. Cytokines and chemokines are key immune mediators that have been related with the pathogenesis of the disease. Obtaining non invasive prognosis markers is a highly desirable objective in order to improve the clinical management of these patients. Patients and methodsIn this study we profiled 17 immune mediators (Th1, Th2, Th17 cytokines, chemokines and growth factors) in the urine of 25 patients with systemic lupus erythematosus with active renal disease by using a Biorad© 17-plex kit on a Luminex© platform. A group of healthy volunteers of similar age and comparable sex distribution was recruited as control (n=10). ResultsResults evidenced that the only detectable mediators in urine were IL-8, MCP-1 and MIP-1b. When levels of these mediators were compared between patients and controls, significantly higher levels of MCP-1 were observed in the urine of the patients. MCP-1 levels in urine correlated positively with the SLEDAI score in a significant way and negatively with plasma levels of complement C4. ConclusionsOur results reinforce the role of MCP-1 in urine as biomarker of disease activity in renal lupus, excluding the detection of other soluble immune mediators such as Th1, Th2, Th17 cytokines and growth factors as suitable markers in this non invasive sample.

Clinical significance of urinary Monocyte Chemoattractant Protein-1 (uMCP-1) in Indian type 2 diabetic patients at different stages of diabetic nephropathy

ObjectiveMonocyte Chemoattractant Protein-1 (MCP-1) is the strongest known chemotactic factor for monocytes and is upregulated in diabetic nephropathy. So measuring urinary MCP-1 is of great significance in the diagnosis and intervention of early diabetic nephropathy. This study aims at determining the levels of urinary MCP-1 (uMCP-1) at different stages of diabetic nephropathy and to see its correlation with other parameters in Indian type2 diabetic subjects. Materials and methodsA total of 64 (M:F; 40:24) type 2 diabetic subjects were divided into three groups based on their renal function and were compared with non-diabetic controls (Group 1) n=20 (M:F; 13:7). The study groups were Group 2 (normoalbuminuria) n=16, Group 3 (microalbuminuria) n=23 and Group 4 (macroalbuminuria) n=25. Demographic, anthropometric and biochemical details were recorded for all the subjects. Urinary MCP-1 levels were measured by using solid phase ELISA method. ResultsMean levels of uMCP-1 in subjects with type 2 diabetes were significantly higher than in controls (p<0.05). The levels of uMCP-1 in type 2 diabetic subjects increased gradually with deteriorating renal function (p=0.006). There was a significant difference in urinary MCP-1 levels between Group 2 and Group 1 (p<0.001). Levels of uMCP-1 were significantly higher in subjects with eGFR <60ml/min compared to eGFR >60ml/min (p=0.008). uMCP-1 levels correlated positively with uACR or uPCR (r=0.551, p<0.0001), urea (r=0.43, p<0.0001) and creatinine (r=0.478, p<0.0001). A negative correlation between uMCP-1 and eGFR (r=−0.338, p=0.006) was noted. ConclusionOur study demonstrated that urinary MCP-1 levels increased gradually in type 2 diabetic subjects with deteriorating renal function. It is significantly associated with the other risk factors of diabetic nephropathy.

Soluble epoxide hydrolase gene deletion attenuates renal injury and inflammation with DOCA-salt hypertension

Inhibition of soluble epoxide hydrolase (sEH) has been shown to be renal protective in rat models of salt-sensitive hypertension. Here, we hypothesize that targeted disruption of the sEH gene (Ephx2) prevents both renal inflammation and injury in deoxycorticosterone acetate plus high salt (DOCA-salt) hypertensive mice. Mean arterial blood pressure (MAP) increased significantly in the DOCA-salt groups, and MAP was lower in Ephx2-/- DOCA-salt (129 +/- 3 mmHg) compared with wild-type (WT) DOCA-salt (145 +/- 2 mmHg) mice. Following 21 days of treatment, WT DOCA-salt urinary MCP-1 excretion increased from control and was attenuated in the Ephx2-/- DOCA-salt group. Macrophage infiltration was reduced in Ephx2-/- DOCA-salt compared with WT DOCA-salt mice. Albuminuria increased in WT DOCA-salt (278 +/- 55 microg/day) compared with control (17 +/- 1 microg/day) and was blunted in the Ephx2-/- DOCA-salt mice (97 +/- 23 microg/day). Glomerular nephrin expression demonstrated an inverse relationship with albuminuria. Nephrin immunofluorescence was greater in the Ephx2-/- DOCA-salt group (3.4 +/- 0.3 RFU) compared with WT DOCA-salt group (1.1 +/- 0.07 RFU). Reduction in renal inflammation and injury was also seen in WT DOCA-salt mice treated with a sEH inhibitor {trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid; tAUCB}, demonstrating that the C-terminal hydrolase domain of the sEH enzyme is responsible for renal protection with DOCA-salt hypertension. These data demonstrate that Ephx2 gene deletion decreases blood pressure, attenuates renal inflammation, and ameliorates glomerular injury in DOCA-salt hypertension.

Contrasting roles of ETA and ETB receptors in angiotensin II‐high salt diet‐induced hypertension

Endothelin (ET) receptor blockade reduces blood pressure and renal injury in angiotensin II(AngII)‐induced hypertension when treatment is initiated at the commencement of AngII infusion. This study tested if ET receptor blockade is also beneficial if treatment is initiated after the onset of hypertension. Male rats received either normal salt diet throughout the experiment (Control), 14‐day AngII infusion (65ng/min s.c.) plus 8% NaCl diet (AngII‐HS) or AngII‐HS plus the ETA receptor antagonist ABT‐627 commencing on day 8 of AngII‐HS treatment (AngII‐HS+ABT). Robust hypertension was observed in AngII‐HS and AngII‐HS+ABT groups by day 7 (mean arterial pressure, MAP 170±5 and 165±5mmHg). Further increase in hypertension was significantly attenuated by treatment with ABT‐627 from day 8 (MAP 168±5 and 191±3mmHg at day 13 in AngII‐HS+ABT and AngII‐HS, P&lt;0.001). Urine was collected for 24 h at day 14. Treatment with ABT‐627 did not blunt the significant increases in urinary protein (19‐fold vs control), albumin (25‐fold) or MCP‐1 excretion (6 to 8‐fold) induced by AngII‐HS treatment. To determine the effect of ETB receptor blockade during AngII‐HS treatment, separate groups of rats were treated with AngII‐HS alone or with the ETB receptor antagonist A‐192621 commencing on day 8 of AngII‐HS treatment. A‐192621 significantly (P&lt;0.05) enhanced AngII‐HS‐induced hypertension by ~14 mmHg over the final 4 days of AngII‐HS treatment but did not affect urinary protein or albumin excretion at day 14. Thus, our data indicate a role for the ETA receptor in the progression of hypertension in this model. However, ETA receptor blockade after hypertension had developed was insufficient to attenuate renal injury, at least over the timeframe studied. ETB receptor blockade exacerbates AngII‐HS hypertension, suggesting that ETB receptor blockade should be avoided in treating hypertension.

Induction of hemeoxygenase‐1 slows the progression of hypertension and proteinuria in spontaneously hypertensive rats

The renoprotective mechanisms of hemeoxygenase‐1 (HO‐1) induction remain unclear. We hypothesize that induction of HO‐1 will decrease blood pressure and proteinuria in spontaneously hypertensive rats (SHR) via antioxidant and anti‐inflammatory mechanisms. Male WKY and SHR were injected with the HO‐1 inducer cobalt protoporphrin (CoPP, 1.5 mg/kg s.c. bi‐weekly) for two weeks. CoPP reduced mean arterial pressure (MAP) (133±2 vs. 144±4 mmHg, p&lt;0.05) and proteinuria (24±2 vs. 14±1mg/day, p&lt;0.05) in SHR rats compared to baseline values, with no effect in WKY. Renal cortical superoxide (O2−) production and urinary 8‐isoprostane excretion were higher in SHR compared to WKY (O2−: 10.5±1 vs.6±1 CPM/mg protein, p&lt;.05; 8‐iso: 7±1.3 vs. 3.4±0.8 ng/day, p&lt;0.05) and CoPP attenuated oxidative stress levels in SHR only (O2−: 5.1±1 CPM/mg, p&lt;0.05; 8‐iso: 4.4±0.7 ng/day). SHR showed a marked elevation in urinary MCP‐1 excretion compared with WKY (19±2 vs. 7.5±2.5 ng/day, p&lt;0.05) and this was attenuated by induction of HO‐1 in SHR (10.5±2.7 ng/day). Plasma nitrite was reduced in SHR compared to WKY (7.9±1.1 vs. 12.7±1.5 μM, p&lt;0.05) and HO‐1 induction increased plasma nitrite levels in SHR (17.5±2.9 μM). These data demonstrate that induction of HO‐1 slows the progression of hypertension and proteinuria in SHR. These changes were associated with reduced renal oxidative stress and inflammation and increased indices of nitric oxide.

Angiotensin II Type 1 Receptor Blockers Reduce Urinary Angiotensinogen Excretion and the Levels of Urinary Markers of Oxidative Stress and Inflammation in Patients with Type 2 Diabetic Nephropathy

To demonstrate that the administration of an angiotensin (Ang) II type 1 receptor (AT1R) blocker (ARB) inhibits the vicious cycle of high glucose (HG)-reactive oxygen species (ROS)-angiotensinogen (AGT)-Ang II-AT1R-ROS by suppressing ROSs and inflammation, thus ameliorating diabetic nephropathy (DN). Thirteen hypertensive DN patients were administered ARBs, and the following parameters were evaluated before and 16 weeks after the treatment: urinary AGT (UAGT), albumin (albumin-creatinine ratio: ACR), 8-hydroxyde-oxyguanosine (8-OHdG), 8-epi-prostaglandin F2alpha(8-epi-PGF2alpha), monocyte chemoattractant protein (MCP)-1, interleukin (IL)-6, and IL-10. ARB treatment reduced the blood pressure and urinary levels of AGT, ACR, 8-OHdG, 8-epi-PGF2alpha, MCP-1, and IL-6 but increased the urinary levels of IL-10. The reduction rate of UAGT correlated with the reduction rate of blood pressure; the reduction rates of the urinary ACR, 8-OHdG, 8-epi-PGF2alpha, MCP-1, and IL-6 levels; and the increase rate of the urinary IL-10 levels. Moreover, subjects who had high UAGT values at baseline exhibited higher reduction rates of urinary albumin excretion. ARB-induced blockade of the abovementioned vicious cycle contributes to the renoprotective effects of ARBs in DN. The urinary levels of AGT could represent a predictive factor for reduced ACR in patients receiving ARB treatment.

Reduction of the vitamin D hormonal system in kidney disease is associated with increased renal inflammation

To examine any potential role for 1,25-dihydroxyvitamin D (1,25(OH)2D) in inflammation associated with chronic kidney disease we measured vitamin D metabolites, markers of inflammation and gene expression in 174 patients with a variety of kidney diseases. Urinary MCP-1 protein and renal macrophage infiltration were each significantly but inversely correlated with serum 1,25(OH)2D levels. Logistic regression analysis with urinary MCP-1 as binary outcome showed that a 10-unit increase in serum 1,25(OH)2D or 25OHD resulted in lower renal inflammation. Analysis of 111 renal biopsies found that renal injury was not associated with a compensatory increase in mRNA for the vitamin D-activating enzyme 25-hydroxyvitamin D-1alpha-hydroxylase (CYP27B1), its catabolic counterpart 24-hydroxylase, or the vitamin D receptor. There was, however, a significant association between tissue MCP-1 and CYP27B1. Patients with acute renal inflammation had a significant increase in urinary and tissue MCP-1, macrophage infiltration, and macrophage and renal epithelial CYP27B1 expression but significantly lower levels of serum 1,25(OH)2D in comparison to patients with chronic ischemic disease despite similar levels of renal damage. In vitro, 1,25(OH)2D attenuated TNFalpha-induced MCP-1 expression by human proximal tubule cells. Our study indicates that renal inflammation is associated with decreased serum vitamin D metabolites and involves activation of the paracrine/autocrine vitamin D system.

The role of capillary density, macrophage infiltration and interstitial scarring in the pathogenesis of human chronic kidney disease

To assess the relationship between interstitial capillary density and interstitial macrophages we prospectively measured these factors in situ in 110 patients with chronic kidney disease. Macrophage numbers and urinary MCP-1/CCL2 levels significantly correlated inversely with capillary density which itself significantly correlated inversely with chronic damage and predicted disease progression. In 54 patients with less than 20% chronic damage, there was a significant correlation between the urinary albumin to creatinine ratio and MCP-1/CCL2, and MCP-1/CCL2 and macrophages but not between MCP-1/CCL2 and capillary density. Conversely, in 56 patients with over 20% chronic damage there was no correlation between MCP-1/CCL2 and macrophages but there were significant inverse correlations between capillary density and both macrophages and chronic damage. The expression of VEGF mRNA significantly correlated with macrophage infiltration, capillary density and chronic scarring. In an ischemic-hypertensive subgroup there was upregulation of the hypoxia marker carbonic anhydrase IX and with over 20% chronic damage an increased macrophage to CCR2 ratio. Our study shows that proteinuria and MCP-1/CCL2 are important for macrophage recruitment in early disease. As renal scarring evolves, alternative pathways relating to progressive tissue ischemia secondary to obliteration of the interstitial capillary bed predominate.

Pioglitazone attenuates diabetic nephropathy through an anti-inflammatory mechanism in type 2 diabetic rats

Peroxisome proliferator-activated receptors (PPARs) are nuclear transcription factors that play a role in insulin sensitivity, lipid metabolism and inflammation. However, the effects of PPARgamma agonist on renal inflammation have not been fully examined in type 2 diabetic nephropathy. In the present study, we investigated the effect and molecular mechanism of the PPARgamma agonist, pioglitazone, on the progression of diabetic nephropathy in type 2 diabetic rats. Inflammatory markers including NF-kappaB, MCP-1 and pro-fibrotic cytokines were determined by RT-PCR, western blot, immunohistochemical staining and EMSA. In addition, to evaluate the direct anti-inflammatory effect of PPARgamma agonist, we performed an in vitro study using mesangial cells. Treatment of OLETF rats with pioglitazone improved insulin sensitivity and kidney/body weight, but had a little effect on blood pressure. Pioglitazone treatment markedly reduced urinary albumin and MCP-1 excretion, and ameliorated glomerulosclerosis. In cDNA microarray analysis using renal cortical tissues, several inflammatory and profibrotic genes were significantly down-regulated by pioglitazone including NF-kappaB, CCL2, TGFbeta1, PAI-1 and VEGF. In renal tissues, pioglitazone treatment significantly reduced macrophage infiltration and NF-kappaB activation in association with a decrease in type IV collagen, PAI-1, and TGFbeta1 expression. In cultured mesangial cells, pioglitazone-activated endogenous PPARgamma transcriptional activity and abolished high glucose-induced collagen production. In addition, pioglitazone treatment also markedly suppressed high glucose-induced MCP-1 synthesis and NF-kappaB activation. These data suggest that pioglitazone not only improves insulin resistance, glycaemic control and lipid profile, but also ameliorates renal injury through an anti-inflammatory mechanism in type 2 diabetic rats.

Lipocalin‐2, TWEAK, and Other Cytokines as Urinary Biomarkers for Lupus Nephritis

Lupus nephritis (LN) is a major cause of morbidity and mortality in systemic lupus erythematosus (SLE). As the course of LN is often unpredictable, it is important to identify reliable, noninvasive methods to repeatedly assess the condition of the kidneys in these patients. Urinary biomarkers are easily obtained and probably are best at reflecting the current renal status, as they specifically represent local inflammatory activity. In this article, we review a number of potential urinary biomarkers, including Lipocalin-2, MCP-1, IP-10, IL-6, and IL-8. Additionally, we provide evidence for the possible utility of the novel cytokine TWEAK as a urinary biomarker for LN.

Urinary levels of RANTES and M-CSF are predictors of lupus nephritis flare

To explore a new predictor of renal flares after successful inductive treatment for diffuse proliferative glomerulonephritis(DPGN) in patients with lupus nephritis. A cohort of patents with SLE DPGN who were treated initially with prednisone and cyclophosphamide were studied. Those who responded to inductive treatment were followed up for the occurrence of renal flares. Urinary levels of RANTES, MCP-1 and M-CSF were measured by ELISA. Other clinical and laboratory data were collected. The predictors and outcome of renal flare were analyzed. Seventy-three qualified patients with SLE DPGN were investigated. After a mean follow-up of 24.5 +/- 6.4 months, 22 patients experienced renal flares. The median time to relapse was 14.1 +/- 4.1 months. The patients experiencing renal flare showed higher urinary RANTES, MCP-1 and M-CSF. However, independent predictors of renal flares were increased urinary RANTES and M-CSF. Eight patents developed doubling of the serum creatinine (CRX2) level. The occurrence of renal flares was the only predictor of CRX2. Persistently increased urinary levels of RANTES and M-CSF after initial remission are predictors of renal flare in patients with SLE DPGN. Our results indicate monitoring urinary pro-inflammatory factors may direct us in managing those patients.

Endotoxin and cisplatin synergistically induce renal dysfunction and cytokine production in mice

A major toxicity of the cancer chemotherapeutic agent cisplatin is acute renal failure. Sepsis is a common cause of acute renal failure in humans and patients who receive cisplatin are at increased risk for sepsis. Accordingly, this study examined the interactions between cisplatin and endotoxin in vivo with respect to renal function and cytokine production. Mice were treated with either a single dose of cisplatin or two doses of LPS administered 24 h apart, or both agents in combination. Administration of 10 mg/kg cisplatin had no effect on blood urea nitrogen or creatinine levels throughout the course of the study. LPS resulted in a modest rise in blood urea nitrogen at 24 and 48 h, which returned to normal by 72 h. In contrast, mice treated with both cisplatin and LPS developed severe renal failure and an increase in mortality. Urine, but not serum, TNF-alpha levels showed a synergistic increase by cisplatin and LPS. Urinary IL-6, MCP-1, KC, and GM-CSF also showed a synergistic increase with cisplatin+LPS treatment. The renal dysfunction induced by cisplatin+LPS was completely dependent on TLR4 signaling and partially dependent on TNF-alpha production. Increased cytokine production was associated with a moderate increase in infiltrating leukocytes which was not different between cisplatin+LPS and LPS alone. These results indicate that cisplatin and LPS act synergistically to produce nephrotoxicity which may involve proinflammatory cytokine production.

The relationship between albuminuria, MCP-1/CCL2, and interstitial macrophages in chronic kidney disease

Glomerular-derived proteins may activate tubular cells to express the macrophage-directed chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2). Macrophages at interstitial sites have a central role in directing renal scarring. We have prospectively assessed the relationship between albuminuria, urinary MCP-1/CCL2, interstitial macrophage infiltration, in situ damage, and clinical outcomes in a large group of patients with chronic kidney disease. We studied 215 patients and quantified albumin-creatinine ratio (ACR), urinary MCP-1/CCL2, interstitial macrophage numbers, and in situ damage. ACR correlated with urinary MCP-1/CCL2 (correlation 0.499; P<0.001), interstitial macrophage numbers (correlation 0.481; P<0.001), and index of chronic damage (correlation 0.363; P<0.001). Macrophage numbers closely correlated with in situ damage (correlation 0.755; P<0.001). By multivariate analysis ACR, urinary MCP-1/CCL2, and interstitial macrophage numbers were interdependent. By Kaplan-Meier survival analysis albuminuria, urinary MCP-1/CCL2, interstitial macrophages, and chronic damage predict the outcome. ACR, macrophage numbers, chronic damage, and creatinine independently predicted renal survival. The association of ACR with other variables was strongest in patients with less advanced disease states. There is a close association between albuminuria, urinary MCP-1/CCL2, and interstitial macrophage infiltration with in situ damage and clinical outcomes. These findings support the hypothesis that albuminuria triggers tubular MCP-1/CCL2 expression with subsequent macrophage infiltration. These processes may represent the dominant pathway for the progression of renal injury before the establishment of advanced renal scarring.

Pentoxifylline ameliorates proteinuria through suppression of renal monocyte chemoattractant protein-1 in patients with proteinuric primary glomerular diseases

Proteinuria (albuminuria) reflects dysfunction of the glomerular permeability barrier in which inflammatory cytokines play a key role. Pentoxifylline (PTX) is a phosphodiesterase inhibitor that possesses potent anti-inflammatory and immunomudulatory effects. This study evaluated the effectiveness of PTX to reduce proteinuria and inflammatory mediators in patients with proteinuric primary glomerular diseases. Seventeen patients with primary glomerular diseases, a persistent spot proteinuria exceeding 1.5 g/g creatinine (Cr) and a glomerular filtration rate between 24 and 115 ml/min/1.73 m(2) were treated with PTX 400 mg twice daily for 6 months. Before and after the treatment, serum Cr, plasma renin activity and aldosterone concentrations, plasma and urinary tumor necrosis factor (TNF)-alpha, interleukin-1beta and monocyte chemoattractant protein (MCP)-1, as well as urinary protein and Cr were measured. PTX significantly reduced urinary protein excretion, along with an increase of serum albumin. A significant correlation existed between the basal urinary protein/Cr and the basal urinary MCP-1/Cr ratios. PTX lowered the urinary MCP-1/Cr ratio, and the percent reduction of urinary protein/Cr ratio correlated directly with the precent decrease of urinary MCP-1/Cr ratio after PTX treatment. There was no significant change in blood pressure, renal function, biochemical parameters, plasma renin activity and aldosterone concentrations, or plasma TNF-alpha and MCP-1 levels during the study. In conclusion, administration of PTX 800 mg per day is safe and effective for reducing proteinuria in patients with proteinuric primary glomerular diseases. This beneficial effect occurs in close association with a reduction of urinary MCP-1 excretion.

Aldosterone Blockade Attenuates Urinary Monocyte Chemoattractant Protein-1 and Oxidative Stress in Patients with Type 2 Diabetes Complicated by Diabetic Nephropathy

Aldosterone causes organic impairment by enhancement of oxidative stress and subsequent induction of proinflammatory cytokines and chemokines. This study was designed to investigate the effect of spironolactone, an aldosterone blocker, on oxidative stress and the level of urinary monocyte chemoattractant protein (MCP)-1, a cysteine-cysteine chemokine that may contribute to progression of various nephropathies in type 2 diabetic patients with diabetic nephropathy. DESIGN, SETTING, PATIENTS AND OTHER PARTICIPANTS, AND INTERVENTION: The patients were randomly assigned to two groups in which they received either spironolactone (50 mg/d; n = 23) or amlodipine (2.5 mg/d; n = 14). Urinary 8-iso-prostaglandin (PG) F2alpha (a marker of oxidative stress), urinary MCP-1, and urinary albumin excretion (UAE) were measured at the start of administration (0 months) and after 3 months in each group. Baseline levels of these variables were also measured in 25 age-matched healthy subjects. There were significant positive correlations between log(10)-transformed (log) 8-iso-PGF2alpha and log MCP-1 levels in control and diabetic subjects and all subjects combined, but no correlations between log UAE and log 8-iso-PGF2alpha or log MCP-1 were found in any group. Significant decreases in 8-iso-PGF2alpha, MCP-1, and UAE were observed with spironolactone (P = 0.0001, P = 0.0041, and P = 0.0037, respectively), and systolic blood pressure significantly decreased after both spironolactone and amlodipine therapy (P = 0.00011 and P = 0.0051, respectively). Our data suggest that urinary MCP-1 is correlated with oxidative stress as measured by urinary 8-iso-PGF2alpha and that spironolactone can decrease urinary MCP-1 and oxidative stress.

Role of aldosterone in diabetic nephropathy

In the last 10 years, many studies have focused on the non-classical action of aldosterone. One of the most important new aspects of aldosterone is its pathogenic role as proinflammatory and profibrotic molecules. It has been reported that aldosterone induces myocardial fibrosis and vascular inflammation through up-regulation of various proinflammatory and profibrotic cytokines. We investigated the effect of aldosterone and spironolactone, which is a non-selective mineralocorticoid receptor antagonist, on monocyte chemoattractant peptide (MCP-1) and collagen synthesis in cultured mesangial and tubular epithelial cells. In addition, to evaluate the effect of spironolactone on diabetic nephropathy, we used Otsuka Long-Evans Tokushima Fatty (OLETF) rats which are known type 2 diabetic animal models. Spironolactone treatment did not induce any significant change in blood glucose levels and blood pressure. However, spironolactone therapy significantly inhibited urinary albumin and MCP-1 excretion. Spironolactone treatment also suppressed renal mRNA expression for MCP-1, macrophage migration inhibitory factor (MIF) as well as intrarenal protein synthesis for ED-1 and MIF. Morphologically, spironolactone treatment significantly prevented glomerulosclerosis, collagen deposition and connective tissue growth factor (CTGF) expression in diabetic rats. In cultured cell experiments, aldosterone directly increased the MCP-1, collagen secretion and spironolactone treatment abolished aldosterone-induced MCP-1 and collagen synthesis. Surprisingly, aldosterone treatment did not induce any significant change in TGFbeta1 gene transcription. Finally, we found that NF-kB activity was increased after stimulation with aldosterone and spironolactone therapy inhibited their activation. In addition, prior treatment with pyrrolidine dithiocarbamate (PDTC), which is a NF-KB inhibitor, inhibited aldosterone-induced MCP-1 protein secretion. These results suggest that aldosterone blockade could play a role in preventing the progression of diabetic nephropathy via anti-inflammatory and antifibrotic mechanisms.

Ureteropelvic Junction Obstruction: Some new Acquisitions about Etiology, Pathophysiology and Diagnostics

The etiopathogenesis of uretero-pelvic junction obstruction (UPJO) has been the subject of many speculations and it remains, in some ways, a debatable matter. Some recently reported thorough research refer to neuro-mediated pathogenetic mechanisms rather than (or together with) myogenic ones. Advances in US, radioisotopic functional imaging, CT and RM contribute to differentiate the obstructive conditions from the non-obstructive ones and to afford today a better assessment of renal functional damage. Particularly, diuretic renography is a non-invasive test for characterization of the renal functional abnormalities resulting from UPJO. Helical CT with angiography is a useful technique for identification of crossing vessels (pyelo-vascular tangle) which can be used for the pre-surgical planning of endopyelotomy. Endoluminal ultrasonography can be used to guide the position of the incision for endopyelotomy (US-guided endopyelotomy). Laboratory examinations are important to determine the overall renal function (serum creatinine, acid-base balance, serum electrolytes, etc.), urinary MCP-1 and NAG (markers of tubular damage), and to rule out urinary tract infections. The management of UPJO (watchful waiting; either open or laparoscopic dismembered pyeloplasty; endoluminal procedures) is greatly influenced by the diagnostic evaluation. The paper aims to outline the advances in both physiopathology and diagnostics of UPJO on the basis of a review of the literature.

MIP-1α and MCP-1 contribute to crescents and interstitial lesions in human crescentic glomerulonephritis

The precise molecular mechanisms of macrophage (Mphi) recruitment and activation in crescentic glomerulonephritis remain to be investigated. We hypothesized that locally produced macrophage inflammatory protein (MIP)-1alpha and monocyte chemoattractant protein (MCP)-1 via the chemokine receptors participate in the pathophysiology of human crescentic glomerulonephritis by recruiting and activating Mphi. We investigated the levels of MIP-1alpha and MCP-1 by enzyme-linked immunosorbent assay (ELISA) in 20 healthy subjects, 20 patients with crescentic glomerulonephritis, and 41 control patients with various other renal diseases. The presence of MIP-1alpha, MCP-1, and the cognate chemokine receptor for MIP-1alpha, CCR5, in the diseased kidneys was evaluated by immunohistochemical and in situ hybridization analyses. MIP-1alpha-positive cells were mainly detected in crescentic lesions, whereas MCP-1 was mainly in the interstitium. In addition, we detected CCR5-positive cells in diseased glomeruli and interstitium. Urinary MIP-1alpha was detected in crescentic glomerulonephritis, even though it was below detectable levels in healthy subjects and in patients with other renal diseases without crescents. Urinary MIP-1alpha levels in the patients with crescentic glomerulonephritis were well correlated with the percentage of cellular crescents and the number of CD68-positive infiltrating cells and CCR5-positive cells in the glomeruli. However, urinary MCP-1 levels were well correlated with the percentage of both total crescents and fibrocellular/fibrous crescents and the number of CD68-positive infiltrating cells in the interstitium. Moreover, elevated urinary levels of both MIP-1alpha and MCP-1 dramatically decreased during glucocorticoid therapy-induced convalescence. These observations suggest that locally produced MIP-1alpha may be involved in the development of cellular crescents in the acute phase via CCR5 and that MCP-1 may be involved mainly in the development of interstitial lesions in the chronic phase when fibrocellular/fibrous crescents are present, possibly through Mphi recruitment and activation.