Zonulin, iron status, and anemia in kidney transplant recipients: are they related?

BACKGROUND: In patients after kidney transplantation, anemia is relatively common and is associated with impaired kidney function, subclinical inflammatory state, and immunosuppressive treatment. Zonulin-prehaptoglobin-2, a newly discovered protein, is necessary for integrity of intracellular tight junctions in the gut. Taking into consideration iron metabolism, including its absorption in the gut, we designed a cross-sectional study to look for the possible interactions among zonulin, iron status, and anemia in kidney transplant recipients. METHODS: The study was performed on 72 stable kidney transplant recipients and 22 healthy volunteers. Zonulin, iron status, and inflammatory markers were assessed with the use of commercially available kits. RESULTS: Zonulin was significantly lower in kidney allograft recipients than in healthy volunteers (P < .001). Zonulin correlated with systolic blood pressure (r = -0.33; P < .05), thyroid-binding globulin (r = 0.24; P < .05), hematocrit (r = 0.28; P < .005), hemoglobin (r = 0.32; P < .01), total protein (r = -0.33; P < .01), erythrocyte count (r = 0.26; P < .05), and fasting glucose (r = -0.25; P < .05). Zonulin was not affected by sex, type of immunosuppressive therapy, presence of diabetes, coronary artery disease, heart failure, hypertension, or cause of end-stage renal disease. Zonulin was not related to any of the iron parameters studied. In multiple regression analysis, predictors of zonulin were total protein and thyroglobulin-binding protein, explaining 46% of variation. CONCLUSIONS: Zonulin, with its poorly defined function, does not seem to play a role in the anemia in kidney allograft recipients; however, it seems to be related to the absorption process in the gut.

Hemojuvelin and iron metabolism in kidney and heart allograft recipients.

INTRODUCTION: Hemojuvelin plays an essential role in the regulation of hepcidin expression, specifically in the iron-sensing pathway. Dietary iron sensing and inflammatory pathways converge in the regulation of the key regulator hepcidin. Hepcidin is a small defensin-like peptide whose production by hepatocytes is modulated in response to anemia, hypoxia, or inflammation. We studied correlations of hemojuvelin with markers of iron status and of inflammation among 61 prevalent kidney allograft recipients and 136 prevalent heart transplant recipients. METHODS: Complete blood count, urea, serum lipids, fasting glucose, and creatinine were measured using standard laboratory methods in the central laboratory of the hospital. Hepcidin, soluble transferin receptor (sTFR), interleukin-6 (IL-6) and hemojuvelin were assayed by enzyme immunosorbent assay using commercially available kits. RESULTS: Among heart transplant recipients hemojuvelin correlated strongly with kidney function, transferrin saturation and white blood cell count; moderately with red blood cell count, hepcidin, IL-6, high-sensitivity C-reactive protein (hsCRP) and weakly with sTfR. Multiple regression analysis revealed the predictors of hemojuvelin to be kidney function and TSAT, explaining 79% of the variations among hemojuvelin values in heart transplant recipients. Among kidney transplant recipients hemojuvelin correlated with kidney function, TSAT, hepcidin and hsCRP, and tended to correlate with IL-6. Predictors of hemojuvelin on multiple regression analysis were TSAT and creatinine. CONCLUSIONS: Elevated hemojuvelin as well as hepcidin levels in kidney or heart transplant recipients may be due to the impaired kidney function and disturbed iron status frequently encountered among this population.

Is hemojuvelin a possible new player in iron metabolism in hemodialysis patients?

INTRODUCTION: Hemojuvelin (HJV) is highly expressed in the liver, skeletal muscles, and heart, seems to play a role in iron absorption and release from cells, and has anti-inflammatory properties. Moreover, HJV plays an essential role in the regulation of hepcidin expression, specifically in the iron-sensing pathway. Hepcidin has emerged as a key regulator of iron homeostasis. In this study we tested for the first time the hypothesis that HJV is related to iron metabolism in hemodialysis (HD) patients. METHODS: Iron status, complete blood count, and serum creatinine, albumin, and lipids were assessed, using standard laboratory methods. Serum levels of soluble transferrin receptor (sTFR), high-sensitivity CRP, IL-6, hepcidin, and HJV were measured using commercially available kits. RESULTS: Serum HJV, hepcidin, ferritin, IL-6, hsCRP, and serum creatinine were significantly higher (all P<0.001), whereas serum iron, sTFR, transferrin, hemoglobin, and erythrocyte count were significantly lower in HD patients, compared to healthy volunteers (all P<0.001). In univariate analysis, HJV was strongly correlated (P<0.001) with ferritin, transferrin saturation, and TIBC, as well as with hsCRP, hepcidin, Kt/V (P<0.01) and residual renal function, the presence of diabetes, APKD, and coronary heart disease. Predictors of HJV level in multiple regression analysis were ferritin (beta value was 0.50, P=0.00004) and transferrin saturation (beta value was 0.47, P=0.0002), explaining 81% of the HJV variations. CONCLUSIONS: Serum HJV is elevated in HD patients and related predominantly to kidney function and iron metabolism. However, HJV is probably not correlated to inflammation. HJV appears to be a new player in iron metabolism in these patients.

Iron metabolism in kidney allograft recipients: still a mystery?

INTRODUCTION: All living organisms have evolved sophisticated mechanisms to maintain appropriate iron levels in their cells and within their body. Recently our understanding of iron metabolism has dramatically increased. Overt labile plasma iron (LPI) represents a component of non-transferrin-bound iron (NTBI) that is both redox active and chelatable, capable of permeating into organs and inducing tissue iron overload. LPI measures the iron-specific capacity of a given sample to produce reactive oxygen species. We studied NTBI correlations with markers of iron status and inflammation in prevalent kidney allograft recipients. METHODS: Complete blood count, urea, creatinine, serum lipids, fasting glucose, ferritin, serum iron, and total iron-binding capacity (TIBC) were studied by standard laboratory method in the hospital central laboratory. NTBI was assessed by FeROS eLPI kit by Aferrix Ltd (Tel Aviv, Israel). A test result of 0.6 U of LPI or more indicated a potential for iron-mediated production of reactive oxygen species in the sample. RESULTS: In kidney transplant recipients NTBI was correlated with TIBC (r=.46, P<.001) and ferritin (r=.31, P<.05), with tendencies to correlate with C-reactive protein. Patients with LPI units≥0.6 showed higher serum iron (P<.05), TIBC (P<.05), ferritin (P<.001) and mean corpuscular volume. High ferritin values together with elevated NTBI content were observed among patients undergoing multiple transfusions before and/or after transplantation. CONCLUSIONS: Elevated NTBI as well as ferritin levels in kidney transplant patients may be due to disturbed iron metabolism, since the human body has no possibility to remove an iron excess. NTBI could be responsible for excessive synthesis of reactive oxygen species. Therefore, it may be linked to complications such as atherosclerosis, which is frequently encountered among this population.

Lack of non-transferrin-bound iron in heart transplant recipients.

BACKGROUND: Labile plasma iron (LPI) is a heterogeneous fraction thought to be composed of iron bound to serum albumin, citrate, and other undefined negatively charged ligands called non-transferrin-bound iron (NTBI). It is associated with formation of reactive oxygen species which are implicated in the pathogenesis of myocardial infarction and bacterial infection. Therefore, the measurement of NTBI could serve as an early marker for reactive oxygen species-induced tissue damage. In this study, we assessed the prevalence of NTBI in heart transplant recipients. METHODS: Complete blood counts, urea, serum lipids, fasting glucose, creatinine, and N-terminal pro-B-type natriuretic peptide were studied by standard laboratory methods in the central laboratory of the hospital. Soluble transferrin receptor was measured using kits from R&D (Abington, UK) and interleukin-6 with kits from Diaclone (Germany). NTBI was assessed in Israel by Aferrix Ltd; LPI≤0.4 units was considered to be negative. RESULTS: In all of the studied patients, NTBI was negative. In the 15 healthy volunteers, all the results were negative. CONCLUSIONS: In heart allograft recipients there is no evidence of reactive oxygen species-induced tissue damage due to either iron overload from oversupplementation or excessive blood transfusion. However, this particular adverse effect should be taken into account when considering treatment of anemia with iron and/or red blood cell transfusions.

Renal nitric oxide production during the early phase of experimental diabetes mellitus.

BACKGROUND: Diabetic nephropathy (DN) is characterized by hyperfiltration and hypertrophy in experimental models of diabetes mellitus (DM). Several studies have demonstrated that the pathophysiologic and morphologic changes in DN are mediated by either an increase or decrease in renal nitric oxide (NO) production and/or activity. The goal of the present study was to determine the effects that the early diabetic state has on NO production in the kidney of rats with streptozotocin-induced DM. METHODS: Experimental DM was induced in rats with streptozotocin. Urinary NO production was measured, and levels and activity of the different NOS isoforms were determined by a combination of techniques, including immunoblotting, immunohistochemistry, diaphorase staining, and reverse transcription-polymerase chain reaction. RESULTS: During the first week of DM, urinary NO metabolites (uNO2 + NO3) were reduced as compared with controls, which were unrelated to changes in serum levels of NO. Total NO synthase (NOS) activity was reduced in the renal cortex beginning at 30 hours after the induction of DM. NADPH diaphorase staining of renal cortical slices showed reduced NOS activity in the macula densa in diabetic animals. By immunohistochemical staining with antibodies to the different isoforms of NOS, it was found that protein levels of the neuroneal NOS (nNOS) isoform was diminished in the macula densa. No changes were found in the levels of endothelial NOS (eNOS) activity and protein in the renal cortex in the early diabetic state. CONCLUSIONS: This study provides strong evidence that renal production of NO is reduced in early DM and that this reduction is associated with decreased levels of nNOS activity and protein in the macula densa.