Furosemide and the Symptom Burden: The Potential Mediating Role of Uremic Toxins in Patients with CKD

Symptoms and Their Correlates in Chronic Kidney Disease

The New Nordic Renal Diet Induces a Pronounced Reduction of Urine Acid Excretion and Uremic Toxins in Chronic Kidney Disease Patients (Stage 3 and 4)

Studies have demonstrated that a low-protein diet supplemented with ketoanalogs (KAs) could significantly retard progression of renal function in patients with chronic kidney disease (CKD) stages 3-5. However, its effects on endothelial function and serum levels of protein-bound uremic toxins remain elusive. Therefore, this study explored whether a low-protein diet (LPD) supplemented with KAs affects kidney function, endothelial function, and serum uremic toxin levels in a CKD-based cohort. In this retrospective cohort, we enrolled 22 stable CKD stage 3b-4 patients on LPD (0.6-0.8 g/day). Patients were categorized into control (LPD only) and study groups (LPD + KAs 6 tab/day). Serum biochemistry, total/free indoxyl sulfate (TIS/FIS), total/free p-cresyl sulfate (TPCS/FPCS), and flow-mediated dilation (FMD) were measured before and after 6 months of KA supplementation. Before the trial, there were no significant differences in kidney function, FMD, or uremic toxin levels between the control and study groups. When compared with the control group, the paired t-test showed a significant decrease in TIS and FIS (all p < 0.05) and a significant increase in FMD, eGFR, and bicarbonate (all p < 0.05). In multivariate regression analysis, an increase in FMD (p < 0.001) and a decrease in FPCS (p = 0.012) and TIS (p < 0.001) remained persistent findings when adjusted for age, systolic blood pressure (SBP), sodium, albumin, and diastolic blood pressure (DBP). LPD supplemented with KAs significantly preserves kidney function and provides additional benefits on endothelial function and protein-bound uremic toxins in patients with CKD.

An accumulation of protein-bound uremic toxins (PBUTs) is one of major reasons for development of uremia-related complications. We examined the PBUT removal ability of a hexadecyl-immobilized cellulose bead (HICB)-containing column for patients undergoing hemodialysis. Adsorption of indoxyl sulfate (IS), a representative PBUT, to HICBs was examined in vitro. The HICB column was used in patients undergoing hemodialysis for direct hemoperfusion with a regular hemodialyzer. The serum IS, indole acetic acid (IAA), phenyl sulfate (PhS), and p-cresyl sulfate (PCS) levels were measured before and after passing the column. HICBs adsorbed protein-free (free) IS in a dose- and time-dependent manner in vitro (55.4 ± 1.4% adsorption of 1 millimolar, 251 µg/mL, IS for 1 h). In clinical studies, passing the HICB-containing column decreased the serum level of free IS, IAA, PhS, and PCS levels significantly (by 34.4 ± 30.0%, 34.8 ± 25.4%, 28.4 ± 18.0%, and 34.9 ± 22.1%, respectively), but not protein-bound toxins in maintenance hemodialysis patients. HICBs absorbed some amount of free PBUTs, but the clinical trial to use HICB column did not show effect to reduce serum PBUTs level in hemodialysis patients. Adsorption treatment by means of direct hemoperfusion with regular hemodialysis may become an attractive blood purification treatment to increase PBUT removal when more effective materials to adsorb PBUTs selectively will be developed.

Aim: To explore the associations between protein-bound uraemic toxins (PBTs), fibre intake and patient-focused outcomes in patients on kidney replacement therapy. Background: Despite removal of small water-soluble uraemic toxins, dialysis patients continue to experience high morbidity and mortality. Recent evidence suggests strong associations between PBTs and poorer patient outcomes and symptom burden. Reducing the generation of PBTs by increasing dietary fibre may be an alternate approach to better patient outcomes. Method: This was a cross-sectional study of haemodialysis (HD), peritoneal dialysis (PD) and kidney transplant patients to determine the associations between uraemic toxins [p-cresyl sulfate (PCS) and indoxyl sulfate (IS)], fibre intake and patient-focused outcomes, incorporating the Integrated Palliative Outcome Scale-Renal (IPOS-renal) and EQ-5D-5L to determine symptom burden and quality of life, while physical capacity was determined using the timed up and go(TUG) test and handgrip strength (HGS). Results: Ninety participants completed the study (n = 30 in each group). There was a correlation between PBTs and the IPOS-renal score, where higher toxin levels were associated with a higher symptom burden. This was the strongest for PCS, where the significance remained after accounting for age and co-morbidities (p < 0.05). Higher PBT levels were also associated with lower HGS (p < 0.05). There was a negative correlation between fibre intake and PBTs, serum PCS (r = −0.36, p < 0.05) and serum IS (r = −0.27, p < 0.05). Lower fibre intake was also associated with a higher symptom burden measured by the IPOS-renal (p < 0.05). Transplant patients consistently performed better, with a reduced symptom burden and improved physical ability compared to dialysis patients. Conclusion: PBTs were associated with symptom burden, and lower physical ability was associated with both PBTs and patient-focused outcomes, and this needs to be further investigated in larger studies.

Accumulation of protein-bound uremic toxins, including indoxyl sulfate and p-cresyl sulfate, are associated with increased cardiovascular disease and mortality in chronic kidney disease (CKD). We performed a systematic review and meta-analysis to synthesize the available strategies for lowering protein-bound uremic toxin levels in CKD patients. We conducted a meta-analysis by searching the databases of MEDLINE, Scopus, and the Cochrane Central Register of Controlled Trials for observational studies and randomized controlled trials (RCTs) that examined the effect of dietary protein restrictions, biotic supplements (including prebiotics, probiotics, and synbiotics), AST-120, dialysis techniques, and the outcome of preservation of residual renal function (RRF) on indoxyl sulfate and p-cresyl sulfate levels. Random-effect model meta-analyses were used to compute changes in the outcomes of interest. A total of 38 articles (2,492 patients), comprising 28 RCTs, 8 single-arm or prospective cohort studies, and 2 cross-sectional studies were included in this meta-analysis. When compared with placebo, prebiotics, synbiotics, and AST-120 provided significantly lower levels of both serum indoxyl sulfate and p-cresyl sulfate. There were no significant reductions in serum indoxyl sulfate and p-cresyl sulfate levels in patients receiving probiotics. Preservation of RRF in dialysis patients resulted in lower levels of both of the protein-bound uremic toxins. When compared with conventional hemodialysis, hemodiafiltration significantly decreased serum p-cresyl sulfate alone, whereas a significant change in serum indoxyl sulfate levels was observed only in studies with long-term observation periods. Very low protein diet (VLPD) and other oral medications yielded insignificant differences in protein-bound uremic toxins. The present meta-analysis demonstrated that prebiotics, synbiotics, and AST-120 can effectively reduce both serum indoxyl sulfate and p-cresyl sulfate in CKD patients when compared with placebo. Preservation of RRF was associated with lower serum indoxyl sulfate and p-cresyl sulfate levels. The effect of biotic supplements was detected only in dialysis patients. For non-dialysis CKD patients, the results were limited due to the small number of studies. Further studies are needed to determine the efficacy in these populations.

Background and Aims In chronic kidney disease (CKD), impaired kidney function results in the accumulation of uremic toxins, which exert deleterious biological effects, contributing to cardiovascular morbidity and mortality. Protein-bound uremic toxins (PBUTs), such as p-cresyl sulfate, indoxyl sulfate and indole-3-acetic acid (IAA), originate from phenolic and indolic compounds, which are end products of the gut bacterial metabolization of aromatic amino acids (AAA). This study investigated the microbial composition in different stages of CKD by isolating, identifying and quantifying PBUT precursor-generating bacteria from fecal samples. Method Using fecal samples from patients in CKD stage 1 (n=6) and stage 5 (n=6), bacteria were cultured in a yeast casitone fatty acid glucose broth medium supplemented with AAA under aerobic (2d at 37°C) and anaerobic conditions (7d at 37°C), and confirmed as PBUT precursor-generating bacteria based on their generation capacity of phenolic and indolic compounds, measured with (U)HPLC. Next, fecal DNA from 14 controls, 111 non-dialyzed and 27 dialyzed CKD patients was used to quantify the total bacterial number but also of 11 of the identified PBUT precursor-generating bacteria with qPCR. Using a Kruskal-Wallis test, bacterial loads were compared between the different CKD stages and control. Correlations between disease stages (control and CKD 1-5) and the abundance of bacterial species were assessed with the Spearman’s rank test. Results In total, 150 different bacterial species were isolated from the CKD fecal samples, of which 101 were identified and 92 classified as PBUT precursor-generating bacteria. In general, p-cresol and phenol were mainly generated under anaerobic conditions, while indole and IAA were generated under both aerobic and anaerobic conditions. Phenolic compounds and IAA were predominantly generated by bacterial species belonging to the Bacteroidaceae, Clostridiaceae, Enterococcaceae and Tannerellaceae, while indolic compounds were mainly generated by Bifidobacteriaceae and Enterobacteriaceae. Quantitative analysis of 11 confirmed PBUT precursor-generating bacteria revealed a higher abundance of Streptococcus spp. and Enterobacteriaceae in fecal samples from HD patients compared to controls and early CKD stages, and for Roseburia spp. compared to CKD 5. Moreover, in HD, the abundance of Clostridioides difficile and Lactobacillus spp. was increased compared to CKD 1-5, and of Escherichia coli compared to control (all p&amp;gt;0.05). The abundance of Bacteroides spp., Faecalibacterium prausnitzii, Akkermansia muciniphila and Bifidobacterium spp. as well as the total number of bacteria was comparable among the different CKD stages and controls. Finally, decrease in kidney function (ranging from control to CKD 5) positively correlated with the abundance of Enterobacteriaceae (rs=0.210), and E. coli (rs=0.286), while an inverse correlation was found with Streptococcus spp. (rs=-0.255), Butyricoccus spp. (rs=-0.326), F. prausnitzii (rs=-0.250), Roseburia spp. (rs=-0.342) and Bifidobacterium spp. (rs=-0.303) (all p&amp;gt;0.05). Conclusion The identified PBUT precursor-generating bacteria are potential targets to reduce the plasma PBUT levels in CKD. In addition, in this CKD cohort, based on qPCR, an altered gut microbial composition with the progression of CKD could be established/confirmed.

Protein-bound uremic toxins (PBUTs) are predominantly excreted by renal tubular secretion and hardly removed by traditional hemodialysis (HD). Accumulation of PBUTs is proposed to contribute to the increased morbidity and mortality of patients with end-stage kidney disease (ESKD). Preserved PBUT excretion in patients with residual kidney function (RKF) and/or increased PBUT clearance with improved dialysis techniques might improve the prognosis of patients with ESKD. The aims of this study are to explore determinants of PBUTs in HD patients, and investigate whether hemodiafiltration (HDF) lowers PBUT plasma concentrations, and whether PBUTs are related to the outcome. Predialysis total plasma concentrations of kynurenine, kynurenic acid, indoxyl sulfate, indole-3-acetic acid, p-cresyl sulfate, p-cresyl glucuronide, and hippuric acid were measured by UHPLC-MS at baseline and after 6 months of follow-up in the first 80 patients participating in the CONvective TRAnsport Study (CONTRAST), a randomized controlled trial that compared the effects of online HDF versus low-flux HD on all-cause mortality and new cardiovascular events. RKF was inversely related to kynurenic acid (p < 0.001), indoxyl sulfate (p = 0.001), indole-3-acetic acid (p = 0.024), p-cresyl glucuronide (p = 0.004) and hippuric acid (p < 0.001) plasma concentrations. Only indoxyl sulfate decreased by 8.0% (−15.3 to 34.6) in patients treated with HDF and increased by 11.9% (−15.4 to 31.9) in HD patients after 6 months of follow-up (HDF vs. HD: p = 0.045). No independent associations were found between PBUT plasma concentrations and either risk of all-cause mortality or new cardiovascular events. In summary, in the current population, RKF is an important determinant of PBUT plasma concentrations in HD patients. The addition of convective transport did not consistently decrease PBUT plasma concentrations and no relation was found between PBUTs and cardiovascular endpoints.

Background/Aims: Accumulation of protein-bound uremic toxins (PBUTs) is associated with mortality due to various systemic disorders in patients with chronic kidney disease (CKD), especially in those undergoing dialysis treatment. The clinical outcomes of such patients could be improved by removing sufficient amounts of PBUTs; however, conventional dialysis lacks this ability. We examined the efficacy of activated carbon in adsorbing circulating PBUTs through direct hemoperfusion (DHP) in vitro. Methods: An in vitro blood circulating system was constructed with 8.5 mL blood circulating around a column containing activated carbon (50, 100, or 200 mg). Bovine blood containing a kind of PBUT (at the same concentration as that found in the blood of dialysis patients) and blood from hemodialysis patients (n = 8) were used. After circulation for the designated amount of time, sera were collected and the levels of PBUTs, including indoxyl sulfate (IS), p-cresyl sulfate, indole acetic acid (IAA), phenyl sulfate, and hippuric acid, were analyzed with mass spectrometry. Results: Activated carbon decreased the PBUT level in bovine blood in a dose-dependent manner (e.g., reduction rate of IS: 67.9 ± 3.8, 83.3 ± 1.9, and 94.5 ± 1.1% after 60-min circulation in columns containing 50, 100, and 200 mg activated carbon respectively). IS, PCS, and IAA were dramatically adsorbed by activated carbon from the blood of patients undergoing hemodialysis (pre vs. post 240-min reaction: IS 2.835 ± 0.876 vs. 0.455 ± 0.108 mg/dL [p < 0.01], PCS 3.208 ± 2.876 vs. 0.768 ± 0.632 mg/dL [p < 0.01], IAA 0.082 ± 0.045 vs. 0.016 ± 0.005 mg/dL [p < 0.01]). Conclusion: Activated carbon effectively adsorbed blood PBUTs in vitro. DHP with activated carbon could be a promising strategy for removing circulating PBUTs from the blood of patients with CKD.

Patients with chronic kidney disease (CKD) often experience a high accumulation of protein-bound uremic toxins (PBUTs), specifically indoxyl sulfate (IS) and p-cresyl sulfate (pCS). In the early stages of CKD, the buildup of PBUTs inhibits bone and muscle function. As CKD progresses, elevated PBUT levels further hinder bone turnover and exacerbate muscle wasting. In the late stage of CKD, hyperparathyroidism worsens PBUT-induced muscle damage but can improve low bone turnover. PBUTs play a significant role in reducing both the quantity and quality of bone by affecting osteoblast and osteoclast lineage. IS, in particular, interferes with osteoblastogenesis by activating aryl hydrocarbon receptor (AhR) signaling, which reduces the expression of Runx2 and impedes osteoblast differentiation. High PBUT levels can also reduce calcitriol production, increase the expression of Wnt antagonists (SOST, DKK1), and decrease klotho expression, all of which contribute to low bone turnover disorders. Furthermore, PBUT accumulation leads to continuous muscle protein breakdown through the excessive production of reactive oxygen species (ROS) and inflammatory cytokines. Interactions between muscles and bones, mediated by various factors released from individual tissues, play a crucial role in the mutual modulation of bone and muscle in CKD. Exercise and nutritional therapy have the potential to yield favorable outcomes. Understanding the underlying mechanisms of bone and muscle loss in CKD can aid in developing new therapies for musculoskeletal diseases, particularly those related to bone loss and muscle wasting.

Chronic kidney disease (CKD) is a highly prevalent condition that is associated with an increased burden of cardiovascular diseases and mortality. Between 7% and 12% of the general population are affected by CKD. In patients with end-stage renal disease (ESRD), the most severe form of CKD, large amounts of uremic toxins are retained in the blood due to impaired kidney function, leading to uremic symptoms and functional as well as biochemical alterations. Currently, extracorporeal blood purification techniques, including hemodialysis, hemofiltration, hemodiafiltration, and hemoperfusion, are widely used to remove small- to medium-sized uremic toxins, maintain homeostasis, and sustain the lives of ESRD patients. However, the available blood purification methods do not efficiently eliminate protein-bound uremic toxins (PBUTs), such as indoxyl sulfate (IS), hippuric acid (HA), p-cresyl sulfate (PCS), and indole-3-acetate (IAA), due to their strong affinity for albumin. The accumulation of PBUTs unfortunately contributes to kidney fibrosis, oxidative stress, cardiovascular events, and poor long-term survival. Therefore, the development of novel blood purification techniques to improve PBUT elimination is of considerable clinical significance. In this review, we first discuss the limitations of current blood purification modalities of PBUT removal and then summarize recent advances in hemoperfusion adsorbents, adsorptive hemodialysis membranes, and dialysate regeneration systems with enhanced PBUT clearance. Finally, we discuss potential challenges and future research directions in this field, with the aim of providing opportunities for more personalized and targeted blood purification therapies for ESRD patients.

In chronic kidney disease, the accumulation of protein-bound uremic toxins (PBUTs), such as hippuric acid (HA), p-cresyl sulfate (PCS) and indoxyl sulfate (IS), contributes to systemic toxicity and organ dysfunction. These toxins bind to plasma proteins, primarily albumin, rendering them resistant to clearance by conventional dialysis. This study hypothesizes that loop diuretics, particularly torasemide and furosemide, can displace PBUTs from their albumin-binding sites, increasing their free fraction and enhancing their removal during hemodialysis. This pilot multicenter crossover study included 17 anuric hemodialysis patients recruited from two hospitals. Participants underwent sequential treatment with furosemide and torasemide, each phase separated by a 1-week washout period. Plasma concentrations of HA, PCS and IS were measured pre- and post-dialysis during baseline (no diuretics) and diuretic treatment phases using high-performance liquid chromatography coupled with tandem mass spectrometry. Changes in pre- and post-dialysis toxin levels were evaluated across treatment phases. Repeated measures analysis of variance assessed the effect of each diuretic treatment on toxin levels and clearance rates. Both loop diuretics increased the free fraction and clearance of PBUTs compared with baseline. Torasemide demonstrated higher efficacy in enhancing the clearance of HA (76.8%) compared with furosemide (63.2%) and baseline (57.3%). For PCS, furosemide achieved greater reductions (66.3%) than torasemide (61.8%) and baseline (24%). Indoxyl sulfate clearance increased significantly with both diuretics (59.1% for furosemide and 58.8% for torasemide) compared with baseline (26.2%). This study demonstrates that loop diuretics, especially torasemide, can enhance the clearance of PBUTs during hemodialysis. Their use mobilizes PBUTs from tissue stores and increases their dialyzability. These findings warrant further investigation in larger, long-term studies to validate the efficacy and clinical benefits of this approach.

Background and Aims Pruritus is a complication that can impair the quality of life of patients with chronic kidney disease (CKD). A previous report has suggested that the severity of pruritus is correlated with high serum concentrations of β2-microglobulin, calcium, and phosphorus, however recent developments in hemodialysis (HD), especially use of high-flux membrane and online hemodiafiltration (OLHDF), may improve the severity of pruritus. Protein-bound uremic toxins (PBUTs), such as indoxyl sulfate and p-cresyl sulfate, are known to be factors associated with atherosclerosis and impaired bone metabolism in CKD and may also be associated with pruritus, especially in those undergoing dialysis treatments. The aim of our study was to assess the severity of pruritus using a new tool, the 5D-itch scale (5D), as well as its relation to circulating PBUTs and dialysis modality in patients with end-stage kidney disease. Method This was a prospective cohort multicenter study, wherein we analyzed the baseline data. The 5D score (5–25 points) was measured in maintenance dialysis patients. The serum concentrations of indoxyl sulfate, p-cresyl sulfate, phenyl sulfate and hippuric acid were measured with mass spectrometry before a dialysis session. The correlations of the 5D scores with serum PBUTs, dialysis modality (HD or OLHDF) and the other background factors were assessed with Spearman correlation coefficients and the generalized linear model. Results This study included 135 dialysis patients (98 HD and 37 OLHDF). The mean 5D score was 6.84 (±SD 3.14). The mean serum concentrations of indoxyl sulfate, p-cresyl sulfate, indole acetic acid, phenyl sulfate, and hippuric acid were 2.99 ± 1.40, 2.05 ± 1.54, 0.11 ± 0.08, 0.94 ± 0.71, and 4.52 ± 3.17 mg/dL, respectively. There was a significant correlation between the 5D score and dialysis modality (correlation coefficient = 0.24, p &amp;lt; 0.01). Dialysis method was associated with the 5D score by the generalized linear model, and OLHDF was associated with a lower 5D score. There were no significant associations between the 5D score and serum concentrations of the various PBUTs (e.g., for indoxyl sulfate, the correlation coefficient was 0.11, p = 0.20) Conclusion Pruritus, as assessed by the 5D score, was associated with dialysis modality, but not with the serum concentration of PBUTs. Further studies will be needed to understand the mechanism of the effect of OLHDF on pruritus as well as the role of uremic toxins in the itchy skin in CKD patients.

Although the relationship between protein-bound uremic toxins (PBUTs) and cardiac structure and cardiac mortality in chronic kidney disease (CKD) has been studied in the past, the association between cardiac dysfunction and PBUTs has not yet been studied. We therefore evaluated the association between impaired peak cardiac performance and the serum free and total concentrations of potentially cardiotoxic PBUTs. In a cross-sectional study of 56 male CKD patients (stages 2–5 (pre-dialysis)) who were asymptomatic with no known cardiac diseases or diabetes we measured peak cardiac power (CPOmax), aerobic exercise capacity (VO2max), and echocardiographic parameters of cardiac morphology and evaluated their association with PBUTs. The serum total and free concentrations of indoxyl sulfate (IXS), p-cresyl sulfate (PCS), p-cresyl glucuronide, indole acetic acid, and hippuric acid showed significant negative correlation with CPOmax and VO2max. IXS and PCS were independently associated with CPOmax and VO2max even after controlling for eGFR. No correlation between left ventricular mass index (LVMI) and PBUTs was seen. The present study for the first time has demonstrated the association between subclinical cardiac dysfunction in CKD and serum levels of a panel of PBUTs. Further studies are required to evaluate the mechanism of cardiotoxicity of the individual uremic toxins.

Background: Propolis possesses many bioactive compounds that could modulate the gut microbiota and reduce the production of uremic toxins in patients with chronic kidney disease (CKD) undergoing hemodialysis (HD). This clinical trial aimed to evaluate the effects of propolis on the gut microbiota profile and uremic toxin plasma levels in HD patients. These are secondary analyses from a previous double-blind, randomized clinical study, with 42 patients divided into two groups: the placebo and propolis group received 400 mg of green propolis extract/day for eight weeks. Indole-3 acetic acid (IAA), indoxyl sulfate (IS), and p-cresyl sulfate (p-CS) plasma levels were evaluated by reversed-phase liquid chromatography, and cytokines were investigated using the multiplex assay (Bio-Plex Magpix®). The fecal microbiota composition was analyzed in a subgroup of patients (n = 6) using a commercial kit for fecal DNA extraction. The V4 region of the 16S rRNA gene was then amplified by the polymerase chain reaction (PCR) using short-read sequencing on the Illumina NovaSeq PE250 platform in a subgroup. Forty-one patients completed the study, 20 in the placebo group and 21 in the propolis group. There was a positive correlation between IAA and TNF-α (r = 0.53, p = 0.01), IL-2 (r = 0.66, p = 0.002), and between pCS and IL-7 (r = 0.46, p = 0.04) at the baseline. No significant changes were observed in the values of uremic toxins after the intervention. Despite not being significant, microbial evenness and observed richness increased following the propolis intervention. Counts of the Fusobacteria species showed a positive correlation with IS, while counts of Firmicutes, Lentisphaerae, and Proteobacteria phyla were negatively correlated with IS. Two months of propolis supplementation did not reduce the plasma levels of uremic toxins (IAA, IS, and p-CS) or change the fecal microbiota.