KIM-1 Levels Research Articles

Suppression of lncRNA GAS6-AS2 alleviates sepsis-related acute kidney injury through regulating the miR-136-5p/OXSR1 axis in vitro and in vivo

Acute kidney injury (AKI) is a common complication of sepsis and increase morbidity and mortality. Long non-coding RNA (LncRNA) GAS6-AS2 was related to inflammation and apoptosis in different diseases by regulating miRNAs and downstream genes, but its role in AKI remains unclear. Thus, we speculated that GAS6-AS2 might function in sepsis-related AKI via regulating target genes. Here, LPS or CLP was used to establish in vitro or in vivo sepsis-related AKI model. The interactions between GAS6-AS2 and miR-136-5p, and miR-136-5p and OXSR1, were validated by luciferase reporter assay, RNA pull-down, or RIP assay. Cell apoptosis was determined by flow cytometry, Western blotting, or IHC. The kidney injury was evaluated by H&E staining. The expression of GAS6-AS2, miR-136-5p, and OXSR1 was determined by qRT-PCR or Western blotting. We found that GAS6-AS2 was up-regulated in LPS-treated HK2 cells and the CLP-induced rat model. In vitro, GAS6-AS2 knockdown decreased cleaved caspase-3 and bax expression and increased bcl-2 expression. The levels of TNF-α, IL-1β, and IL-6 were reduced by GAS6-AS2 down-regulation. GAS6-AS2 knockdown ameliorated oxidative stress in the cells, as indicated by the reduced ROS and MDA levels and the elevated SOD level. In vivo, GAS6-AS2 down-regulation decreased urinary NGAL and Kim-1 levels and serum sCr and BUN levels, and H&E proved that the kidney injury was alleviated. GAS6-AS2 knockdown also reduced apoptosis, inflammation, and oxidation induced by CLP in vivo. Mechanically, GAS6-AS2 sponged miR-136-5p which targeted OXSR1. Overall, lncRNA GAS6-AS2 knockdown has the potential to ameliorate sepsis-related AKI, and the mechanism is related to miR-136-5p/OXSR1 axis.

Evaluation of renal microperfusion in hyperuricemic nephropathy by contrast-enhanced ultrasound imaging

ABSTRACTDiagnostic tools for the early detection of renal injury caused by hyperuricemia are still lacking. Here, we investigated whether contrast-enhanced ultrasound (CEUS) could be used as a diagnostic tool for hyperuricemic nephropathy (HN). In the HN rat model, CEUS detected a significant decline in renal cortical perfusion compared with that in control rats. Peak intensity (PI) values correlated significantly with serum KIM-1 levels and fibrosis scores in HN rats. An early decline in PI values was also observed in chronic kidney disease (CKD) stage 1 patients with HN compared with the controls (61.1±4.52 dB versus 65.80±7.10 dB) and correlated with renal function in the patients with HN. In contrast, an increase in time to reach PI values was detected in HN patients with stage 1 CKD (15.14±1.75 s versus 14.52±4.75 s) and was more pronounced in CKD stage 4 patients (67.32±3.29 s). CEUS was able to detect abnormal renal perfusion in early CKD with HN, which correlated with renal function decline, suggesting that CEUS could be used as a noninvasive tool for assessing renal function in patients with HN.

Ginsenoside Rg1 ameliorates sepsis-induced acute kidney injury by inhibiting ferroptosis in renal tubular epithelial cells.

Acute kidney injury (AKI) represents a prevailing complication of sepsis, and its onset involves ferroptosis. Ginsenoside Rg1 exerts a positive effect on kidney diseases. This study explored the action of ginsenoside Rg1 in sepsis-induced AKI (SI-AKI) by regulating ferroptosis in renal tubular epithelial cells (TECs). Sepsis rat models were established using cecal ligation and puncture (CLP) and cell models were established by treating human renal TECs (HK-2) with LPS to induce ferroptosis. Serum creatinine (SCr) and blood urea nitrogen (BUN) and urine KIM1 contents in rats were determined by ELISA kits. Kidney tissues were subjected to immunohistochemical and H&E stainings. Iron concentration, malondialdehyde (MDA), glutathione (GSH), and ferroptosis-related protein (ferritin light chain [FTL], ferritin heavy chain [FTH], GSH peroxidase 4 [GPX4], and Ferroptosis suppressor protein 1 [FSP1]) levels in kidney tissues and HK-2 cells were measured using ELISA kits and Western blotting. HK-2 cell viability was detected by cell counting kit-8, and cell death was observed via propidium iodide staining. Reactive oxygen species accumulation in cells was detected using C11 BODIPY 581/591 as a molecular probe. In CLP rats, ginsenoside Rg1 reduced SCr, BUN, KIM1, and NGAL levels, thus palliating SI-AKI. Additionally, ginsenoside Rg1 decreased iron content, FTL, FTH, and MDA levels, and elevated GPX4, FSP1, and GSH levels, thereby inhibiting lipid peroxidation and ferroptosis. Moreover, FSP1 knockdown annulled the inhibition of ginsenoside Rg1 on ferroptosis. In vitro experiments, ginsenoside Rg1 raised HK-2 cell viability and lowered iron accumulation and lipid peroxidation during ferroptosis, and its antiferroptosis activity was dependent on FSP1. Ginsenoside Rg1 alleviates SI-AKI, possibly resulting from inhibition of ferroptosis in renal TECs through FSP1.

Scutellarin protects the kidney from ischemia/reperfusion injury by targeting Nrf2.

Acute kidney injury (AKI) results in high morbidity and mortality among inpatients, while effective treatment and intervention are still absent. Therefore, this study aims to explore the effects of Scutellarin (Scu) in experimental models in vivo and in vitro. In vivo experiment, we employed a total of 30 Wistar rats, which further were modelled by a bilateral renal pedicle clip for 45 min, then received 50 mg/kg/day Scu. In vitro, HK-2 cells were administered with 20μΜ Scu and then incubated in hypoxia/reoxygenation (H/R) conditions for 24 h. The knockdown of Nrf2 expression was conducted by small interfering RNA (siRNA) transfection. As a result, the AKI model was well established with an increased SCr, BUN, KIM-1 level, and histological injury score, while Scu treatment reduced the levels above and increased the antioxidative enzyme HO-1. H/R induced an increase of ROS in HK-2 cells, while Scu decreased the ROS level. Bioinformatics results showed the transcription factor Nrf2 was a hub protein during the AKI, which also bound to Scu with low binding energy, indicating that the downstream effect of Scu might be mediated by Nrf2. To verify the suppose above, we employed siRNA against Nrf2, which shows a significant increase in ROS after Nrf2 was blocked. Meanwhile, the HO-1 showed similar expression compared with the 'H/R + Nrf2 siRNA' and 'H/R + Nrf2 siRNA + Scu' group, implying the protective effect of Scu was mediated by the Nrf2/HO-1 pathway. Scu led to up-regulation of HO-1 through activating the Nrf2 signalling pathway, protecting the kidneys from ischemia/reperfusion (I/R)-induced oxidative damage.

Ginsenoside Rg1 attenuates LPS-induced chronic renal injury by inhibiting NOX4-NLRP3 signaling in mice.

Chronic renal injury (CRI) is a common pathological damage in chronic renal disease, and the therapeutic options for preventing its progression are limited at present. Ginsenoside Rg1 (Rg1) is reported to have a protective effect on renal injury by improving oxidative stress and inflammation. Lipopolysaccharide (LPS) plays important roles in inducing inflammatory and high-dose LPS is often used to perform acute renal injury. However, little is known about the effect of low-dose LPS on CRI, and the protective effect of Rg1 against chronic LPS-induced CRI. Here, we reported the protective effect and mechanism of Rg1 against LPS-induced CRI in mice. In this study, the results demonstrated that low-dose LPS (0.25mg/kg) exposure for 14 days significantly induced renal function impairment and renal injury and fibrosis. Meanwhile, LPS exposure significantly increased reactive oxygen species (ROS) generation, NADPH oxidase 4 (NOX4) and NLRP3 inflammasome expression in renal cortex. However, treatment with Rg1, tempol (a superoxide dismutase mimetic), and apocynin (a NOX inhibitor) significantly improved renal function impairment and renal fibrosis, and significantly decreased the levels of TGF-β, IL-1β, KIM-1, β-Gal, and collagen IV in the kidneys. And Rg1 treatment also significantly reduced ROS generation and inhibited the activation of NOX4 and NLRP3 inflammasome. Overall, these results suggest that Rg1 treatment can ameliorate LPS-induced chronic kidney injury and renal fibrosis, the mechanisms may be involved in reducing NOX2-mediated oxidative stress and inhibiting NLRP1 inflammasome.

Glabridin, a bioactive component of licorice, ameliorates diabetic nephropathy by regulating ferroptosis and the VEGF/Akt/ERK pathways

BackgroundGlabridin (Glab) is a bioactive component of licorice that can ameliorate diabetes, but its role in diabetic nephropathy (DN) has seldom been reported. Herein, we explored the effect and underlying mechanism of Glab on DN.MethodsThe bioactive component-target network of licorice against DN was by a network pharmacology approach. The protective effect of Glab on the kidney was investigated by a high-fat diet with streptozotocin induced-diabetic rat model. High glucose-induced NRK-52E cells were used for in vitro studies. The effects of Glab on ferroptosis and VEGF/Akt/ERK pathways in DN were investigated in vivo and in vitro using qRT-PCR, WB, and IHC experiments.ResultsBioinformatics analysis constructed a network comprising of 10 bioactive components of licorice and 40 targets for DN. 13 matching targets of Glab were mainly involved in the VEGF signaling pathway. Glab treatment ameliorated general states and reduced FBG, HOMA-β, and HOMA-insulin index of diabetic rats. The renal pathological changes and the impaired renal function (the increased levels of Scr, BUN, UREA, KIM-1, NGAL, and TIMP-1) were also improved by Glab. Moreover, Glab repressed ferroptosis by increasing SOD and GSH activity, and GPX4, SLC7A11, and SLC3A2 expression, and decreasing MDA and iron concentrations, and TFR1 expression, in vivo and in vitro. Mechanically, Glab significantly suppressed VEGF, p-AKT, p-ERK1/2 expression in both diabetic rats and HG-induced NRK-52E cells.ConclusionsThis study revealed protective effects of Glab on the kidney of diabetic rats, which might exert by suppressing ferroptosis and the VEGF/Akt/ERK pathway.

Urinary biomarkers indicate pediatric renal injury among rural farming communities in Sri Lanka

Pediatric renal injury is an emerging health concern in communities affected by chronic kidney disease of uncertain etiology (CKDu). Early detection of susceptibilities through highly sensitive and specific biomarkers can lead to effective therapeutic and preventive interventions against renal diseases. Here, we aimed to investigate the utility of kidney injury molecule (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL) in early detection of renal abnormalities in selected pediatric communities in Sri Lanka. The study areas were stratified as CKDu endemic, emerging, and non-endemic based on the prevalence of CKDu, and a total of 804 school students (10–18 years of age) participated in the study. The median (IQR) urinary KIM-1 levels of the participants were 0.193 (0.026–0.338), 0.082 (0.001–0.220) and 0.040 (0.003–0.242) ng/mgCr for CKDu endemic, emerging and non-endemic regions respectively. Participants from CKDu endemic regions reported elevated (p < 0.0001) urinary KIM-1 expression compared to those from the other regions. The median (IQR) NGAL levels in participants from CKDu endemic (2.969; 1.833–5.641), emerging (3.374; 1.766–6.103), and non-endemic (3.345; 1.742–5.128 ng/mgCr) regions showed no significant difference. Also, urinary albumin-creatinine ratio (UACR) showed no significant differences across gender or residency. The prevalence of albuminuria was 1–2% in the locations irrespective of CKDu burden. Albuminuric participants reported higher (p < 0.05) urinary KIM-1 levels in comparison to normoalbuminuric participants. Significantly elevated urinary KIM-1 expression in a pediatric population from CKDu affected regions, especially in the presence of albuminuria, may indicate low-grade early renal damage supporting the utility of KIM-1 as a quantifiable biomarker.

Single Cell Dissection of Epithelial-Immune Cellular Interplay in Acute Kidney Injury Microenvironment.

BackgroundUnderstanding the acute kidney injury (AKI) microenvironment changes and the complex cellular interaction is essential to elucidate the mechanisms and develop new targeted therapies for AKI.MethodsWe employed unbiased single-cell RNA sequencing to systematically resolve the cellular atlas of kidney tissue samples from mice at 1, 2 and 3 days after ischemia-reperfusion AKI and healthy control. The single-cell transcriptome findings were validated using multiplex immunostaining, western blotting, and functional experiments.ResultsWe constructed a systematic single-cell transcriptome atlas covering different AKI timepoints with immune cell infiltration increasing with AKI progression. Three new proximal tubule cells (PTCs) subtypes (PTC-S1-new/PTC-S2-new/PTC-S3-new) were identified, with upregulation of injury and repair-regulated signatures such as Sox9, Vcam1, Egr1, and Klf6 while with downregulation of metabolism. PTC-S1-new exhibited pro-inflammatory and pro-fibrotic signature compared to normal PTC, and trajectory analysis revealed that proliferating PTCs were the precursor cell of PTC-S1-new, and part of PTC-S1-new cells may turn into PTC-injured and then become fibrotic. Cellular interaction analysis revealed that PTC-S1-new and PTC-injured interacted closely with infiltrating immune cells through CXCL and TNF signaling pathways. Immunostaining validated that injured PTCs expressed a high level of TNFRSF1A and Kim-1, and functional experiments revealed that the exogenous addition of TNF-α promoted kidney inflammation, dramatic injury, and specific depletion of TNFRSF1A would abrogate the injury.ConclusionsThe single-cell profiling of AKI microenvironment provides new insight for the deep understanding of molecular changes of AKI, and elucidates the mechanisms and developing new targeted therapies for AKI.

MO067: Role of ANT2 in the Development of Obesity-Induced Chronic Kidney Disease

Abstract BACKGROUND AND AIMS Obesity, a widespread condition with an ever-increasing prevalence, imposes a great medical concern since it serves as a risk factor for a wide range of conditions, including the development of chronic kidney disease (CKD). Lipid accumulation and impaired fatty acid β-oxidation in the renal proximal tubular cells (RPTCs) as well as an overall mitochondrial dysfunction have been shown to induce kidney injury, inflammation and fibrosis, leading to the development of CKD. However, the molecular mechanisms linking mitochondrial dysfunction in RPTCs to obesity-induced CKD are not fully understood and require further examination. Adenine nucleotide translocases (ANTs), which transport ADP and ATP through the mitochondrial inner membrane, play an essential role in energy metabolism of eukaryotic cells. ANT2 (Slc25a5) mediates fatty acid-induced uncoupled respiration, and its complete deletion is embryonic lethal. Here we explore its specific role in RPTCs in the development of obesity-induced CKD. METHOD ANT2 protein levels were measured using western blot and immunohistochemistry analyses. RPTC-ANT2-/- animals were generated using the Cre/loxP system. The null mice and their wild-type littermate controls were fed with either a high-fat diet (HFD) or a standard diet (STD) for 24 weeks. Kidney function and lipid content were assessed by biochemical analysis. Histological techniques were used to visualize the morphological changes in the kidney as well as fibrogenesis. Kidney injury and inflammatory markers were measured via qPCR and ELISAs. Mitochondrial function was tested in primary mouse RPTCs extracted from the null and WT animals by measuring oxygen consumption rate via Seahorse as well as reactive oxygen species (ROS) production by DCFDA staining. RESULTS As evident from the double-stained immunofluorescence staining of mouse kidney, high levels of ANT2 were found in RPTCs. Interestingly, its protein expression was downregulated in HFD-fed WT mice. Nullification of ANT2 specifically in RPTCs prevented the obesity-induced kidney dysfunction as reflected by lowered albumin-to-creatinine ratio. This was associated with the improved renal morphology in HFD-fed RPTC-ANT2-/- mice, evident by the reduction in obesity-related increased glomerular and Bowman's space areas and mesangial expansion. Compared with their littermate controls, obese RPTC-ANT2-/- mice showed reduced renal KIM-1 and NGAL protein levels and downregulation in the expression levels of Mcp1 and Lcn2, markers of renal injury and inflammation, respectively. The null mice were also protected from obesity-induced fibrosis, measured by trichrome staining, as well as accumulation of triglycerides and cholesterol in the kidney. In addition, preserved mitochondrial function was found in primary mouse RPTCs lacking ANT2, measured by increased spare respiratory capacity and decreased ROS production. Lastly, downregulated Hif1α gene and protein expression levels were found in RPTCs and kidney tissue lacking ANT2. CONCLUSION Our findings introduce ANT2 in the RPTCs as a key player in the development of obesity-induced CKD. Deletion of RPTC-ANT2 protects the kidney from the deleterious effects of lipotoxicity. Therefore, targeted manipulation of renal mitochondrial metabolism, particularly via inhibiting ANT2, may represent a novel approach for the treatment of obesity-induced CKD.

NGAL and KIM-1 – early urinary biomarkers of nephrotoxicity mediated by cisplatin: Observational study

Background. Cisplatin is widely used in modern oncological practice. Despite high efficacy treatment with cisplatin is conjugated with high risk of nephrotoxicity. Approximately one third of patients develop renal disfunction after first injection of cisplatin. In clinical practice serum creatinine elevation is used as a marker of renal damage, which is observed after failure of 50% of kidney function. That is why the finding of early biomarker of nephrotoxicity is still an issue. NGAL and KIM-1 are markers of renal damage, the predictive value of which has been described in cardiac surgery and resuscitation practice: an increase in the concentration of these markers in urine precedes the development of renal damage, both ischemic and direct toxic.&#x0D; Aim. To evaluate the role of NGAL and KIM-1 in urine as early markers of cisplatin nephrotoxicity.&#x0D; Materials and methods. The study included 50 patients treated with cisplatin in combination with fluoropyrimidines or paclitaxel. Prior to treatment and over a period of 8 weeks, the Friedman test was used to assess blood pressure, plasma creatinine, potassium, urea levels, daily proteinuria, urine NGAL and KIM-1 levels, and glomerular filtration rate (GFR). ROC analysis was used to assess the prognostic significance of NGAL and KIM-1 in the development of nephrotoxicity.&#x0D; Results. There was a statistically significant increase in the level of urea (=17.7; df 4, p=0.001), potassium (=42; df 4, p0.001), a decrease in GFR (=32.3; df 4, p0.001), the appearance of proteinuria (=50.4; df 4, p0.001). The concentration of NGAL and KIM-1 increased already one week after the start of cisplatin therapy, reaching a maximum by 8 weeks (=200; df 4, p0.001). The appearance of NGAL at a concentration of 10.743 ng/ml and KIM-1 at a concentration of 182.4 pg/ml in the first week after administration of cisplatin allows predicting the development of nephrotoxicity by the 8th week with high sensitivity (90.91%) and specificity (94.87%), AUC 0.96.&#x0D; Conclusion. The appearance of NGAL and KIM-1 in urine already in the first week of treatment allows predicting the development of nephrotoxicity a decrease in GFR of less than 60 ml/min by the 8th week of therapy with high sensitivity and specificity. Both biomarkers can be considered early prognostically significant.

Nephrotoxicity evaluation and proteomic analysis in kidneys of rats exposed to thioacetamide

Thioacetamide (TAA) was administered orally at 0, 10, and 30 mg/kg body weight (BW) daily to Sprague–Dawley rats aged 6–7 weeks for 28 consecutive days. Nephrotoxicity and proteomics were evaluated in the kidneys of rats exposed to TAA. The BW decreased, however, the relative kidneys weight increased. No significant histopathologic abnormalities were found in the kidneys. The numbers of monocytes and platelets were significantly increased. However, the mean corpuscular volume and hematocrit values were decreased significantly in rats exposed to 30 mg/kg BW TAA. The expression levels of Kim-1 and NGAL were increased 4 to 5-fold in the kidneys, resulting in significant nephrotoxicity. Proteomic analysis was conducted and a total of 5221 proteins spots were resolved. Of these, 3 and 21 protein spots were up- and downregulated, respectively. The validation of seven proteins was performed by Western blot analysis. The expression level of ASAP2 was significantly upregulated, whereas RGS14, MAP7Dl, IL-3Rα, Tmod1, NQO2, and MUP were reduced. Sixteen isoforms of MUP were found by the 2DE immunoblot assay and were significantly downregulated with increasing exposure to TAA. MUP isoforms were compared in the liver, kidneys, and urine of untreated rats and a total of 43 isoforms were found.

Cerium oxide nanoparticles attenuate the renal injury induced by cadmium chloride via improvement of the NBN and Nrf2 gene expressions in rats.

Cadmium (Cd) is a highly toxic heavy metal that adversely affects both human and animal health. Chronic cadmium exposure causes serious kidney damage. The current study investigated the protective role of cerium oxide nanoparticles (CeO2NPs) against cadmium chloride (CdCl2)-induced renal injury. One hundred and twenty male albino rats were divided into 6 equal groups. Group (C): considered as control group which was given distilled water orally. Group (NC.1 and NC.5): rats were injected i.p. with nanoceria at a dose of (0.1 and 0.5mg/kg b.wt), respectively, twice a week for 2weeks starting at the 15th day of the study. Group (Cd): rats were received CdCl2 orally (10mg/kg b.wt) daily for 28days. Groups (Cd+NC.1 and Cd+NC.5): rats were given CdCl2 orally (10mg/kg b.wt) for 28days and CeO2NPs by i.p. injection at a dose of (0.1 and 0.5mg/kg b.wt), respectively, twice a week for 2weeks started at the 15th day of the experiment. The Cd group exhibited a significant increase in the serum levels of IL-1β, KIM-1, Cys-C, and β2-MG, downregulation of the antioxidant initiator genes such as Nrf-2, and up-regulation of apoptosis markers such as nibrin gene (NBN). Urine examination showed a high level of microalbuminuria, abnormal physical, chemical, and microscopical changes in comparison with control groups. Remarkably, posttreatment with CeO2NPs showed significant improvement in kidney histopathological picture and relieved the alterations in kidney biomarkers, inflammatory markers, urine abnormalities, and expressions of different genes as Nrf-2 and NBN.

The Urinary Level of Injury Biomarkers Is Not Univocally Reflective of the Extent of Toxic Renal Tubular Injury in Rats.

Nephrotoxicity is a major cause of intrinsic acute kidney injury (AKI). Because renal tissue damage may occur independently of a reduction in glomerular filtration rate and of elevations in plasma creatinine concentration, so-called injury biomarkers have been proposed to form part of diagnostic criteria as reflective of tubular damage independently of renal function status. We studied whether the urinary level of NGAL, KIM-1, GM2AP, t-gelsolin, and REGIIIb informed on the extent of tubular damage in rat models of nephrotoxicity, regardless of the etiology, moment of observation, and underlying pathophysiology. At a time of overt AKI, urinary biomarkers were measured by Western blot or ELISA, and tubular necrosis was scored from histological specimens stained with hematoxylin and eosin. Correlation and regression studies revealed that only weak relations existed between biomarkers and tubular damage. Due to high interindividual variability in the extent of damage for any given biomarker level, urinary injury biomarkers did not necessarily reflect the extent of the underlying tissue injury in individual rats. We contended, in this work, that further pathophysiological contextualization is necessary to understand the diagnostic significance of injury biomarkers before they can be used for renal tubular damage severity stratification in the context of nephrotoxic and, in general, intrinsic AKI.

P07 Toxicokinetic profile of VRP-034

BackgroundPolymyxin B (PMB) can be an effective option for treating resistant Gram-negative pathogens, however, its use is severely compromised by its’ dose-limiting toxicity. VRP-034, a novel formulation of PMB, has previously reported a favourable safety profile compared with marketed PMB (Roy et al.1). This study was conducted to understand the toxicokinetic profile of VRP-034 vis-à-vis marketed PMB.Materials and methods30 healthy Sprague-Dawley rats (divided into six groups of n = 5) received one of three doses (3, 9 and 18 mg/kg/day) of either marketed PMB or VRP-034, administered subcutaneously q8 h over 2 days. Blood samples were collected prior to initiation of experiment and at 0.5, 1, 2, 4, 6, 8, 12 and 24 h after the last dose. Plasma concentrations of PMB were quantified by LC-MS. Additionally, renal toxicity was assessed via clinical pathology, kidney injury biomarkers, oxidative stress markers (3-nitrotyrosine and renal tissue lactate), gross and histopathologic observations of kidney. Urine and blood samples were collected at baseline and Day 2 for the assessment of kidney injury (KIM-1, cystatin-C, BUN and serum creatinine) using respective biochemical or ELISA kits. All values are expressed as mean ± SEM.ResultsThe plasma levels of PMB were found to increase in dose-dependent manner in both the groups. At 3 and 9 mg/kg/day, the plasma PMB exposure were similar for both VRP-034 and marketed PMB groups (AUC0–24: 6.47 ± 0.41 μg/mL·h versus 5.66 ± 0.24 μg/mL·h, P>0.05 and 22.90 ± 1.96 μg/mL·h versus 26.84 ± 1.43 μg/mL·h, P>0.05). At the highest dose of 18 mg/kg/day, the AUC0–24 was higher in the VRP-034 group (AUC0–24: 97.83 ± 12.95 μg/mL·h versus 70.48 ± 4.22 μg/mL·h), however this increase was not statistically significant (P>0.05). The levels of early kidney injury urinary biomarkers increased with dose in both the groups (Table 1). On Day 2, Kim-1 levels in the marketed PMB group increased from baseline by 3.96-fold (versus 2.31-fold in VRP-034 group), 8.21-fold (versus 5.41-fold) and 23.41-fold (versus 9.51-fold) at 3, 9, 18 mg/kg/day dose levels respectively. Likewise, cystatin-C levels increased by 6.22-fold (versus 3.26-fold in VRP-034 group), 6.25-fold (versus 3.67-fold) and 13.57-fold (versus 4.33-fold), respectively, at the three dose levels tested. Oxidative stress markers, tissue lactate and 3-nitrotyrosine, were significantly lower (P<0.05) for VRP-034 group compared with marketed PMB group at the highest dose. Histopathology revealed EGTI grade 3 kidney damage in the marketed PMB group compared with grade 1 damage in the VRP-034 group.Table 1.Kidney injury and oxidative stress markers for marketed PMB and VRO-034 groups All values are reported as mean ± SEM.Lactate marker was only measured on Day 2.UCr, urine creatinine.ConclusionsVRP-034 exhibited a more promising toxicokinetic profile as compared with marketed PMB at all dose levels. Our future work will explore the clinical translation of these findings in humans.

Protective antioxidative and anti-inflammatory actions of β-caryophyllene against sulfasalazine-induced nephrotoxicity in rat.

The pathogenesis of sulfasalazine (SFZ)-induced nephrotoxicity is unclear. Moreover, there are no reports on the protective effects of β-caryophyllene (BCP) against SFZ-induced renal injury. Hence, in this study, we measured several oxidative stress and inflammatory regulatory molecules alongside the effects of BCP in SFZ-intoxicated rats. Male rats (n = 48) were distributed to six equal groups as follows: negative control (NC), normal rats treated with low (N-LD; 200 mg/kg/day) and high (N-HD; 400 mg/kg/day) BCP doses, and animals treated with SFZ individually (PC; 600 mg/kg/day) or combined with BCP low (P-LD) and high (P-HD) doses. All drugs were administrated for 14 consecutive days. The NC, N-LD, and N-HD groups showed comparable renal histology and biochemistry. In contrast, abnormal histology, and increased creatinine and urea alongside oliguria and proteinuria were detected in the PC group. Renal specimens from the PC group revealed increased levels of nuclear factor-kappa B (NF-κB), transforming growth factor (TGF)-β with kidney injury molecule (KIM)-1, while the levels of nuclear factor erythroid 2-related factor 2 (Nrf2), AMP-activated protein kinase (AMPK), and protein kinase B (AKT) declined, relative to controls. The PC renal tissue also had markedly higher levels of inflammatory cytokines (tumor necrosis factor [TNF]-α/interleukin [IL]-1β/IL-6) and pro-oxidants (malondialdehyde [MDA]/H2O2/protein carbonyls), whereas those of antioxidants (glutathione [GSH]/glutathione peroxidase [GPx]/superoxide dismutase-1 [SOD1]/catalase [CAT]) and IL-10 decreased and were associated with marked apoptosis. Both BCP regimens ameliorated renal functions and histology, and reduced NF-κB, TGF-β, and KIM-1 levels in addition to those of oxidative stress and inflammation markers. Both protocols also augmented Nrf2, AMPK, AKT, antioxidants, and IL-10. However, P-HD showed better alleviating effects than the N-HD group. In conclusion, this study is the first to link NF-κB, TGF-β, Nrf2, AMPK, and AKT with SFZ-induced nephrotoxicity. In addition, this is the first report to reveal antioxidative and anti-inflammatory effects for BCP against SFZ-associated nephropathy.

Hemodynamics and Metabolic Parameters in Normothermic Kidney Preservation Are Linked With Donor Factors, Perfusate Cells, and Cytokines.

Kidney transplantation is the best renal-replacement option for most patients with end-stage renal disease. Normothermic machine preservation (NMP) of the kidney has been studied extensively during the last two decades and implemented in clinical trials. Biomarker research led to success in identifying molecules with diagnostic, predictive and therapeutic properties in chronic kidney disease. However, perfusate biomarkers and potential predictive mechanisms in NMP have not been identified yet. Twelve discarded human kidneys (n = 7 DBD, n = 5 DCD) underwent NMP for up to 24 h. Eight were perfused applying urine recirculation (URC), four with replacement of urine (UR) using Ringer's lactate. The aim of our study was to investigate biomarkers (NGAL, KIM-1, and L-FABP), cells and cytokines in the perfusate in context with donor characteristics, perfusate hemodynamics and metabolic parameters. Cold ischemia time did not correlate with any of the markers. Perfusates of DBD kidneys had a significantly lower number of leukocytes after 6 h of NMP compared to DCD. Arterial flow, pH, NGAL and L-FABP correlated with donor creatinine and eGFR. Arterial flow was higher in kidneys with lower perfusate lactate. Perfusate TNF-α was higher in kidneys with lower arterial flow. The cytokines IL-1β and GM-CSF decreased during 6 h of NMP. Kidneys with more urine output had lower perfusate KIM-1 levels. Median and 6-h values of lactate, arterial flow, pH, NGAL, KIM-1, and L-FABP correlated with each other indicating a 6-h period being applicable for kidney viability assessment. The study results demonstrate a comparable cytokine and cell profile in perfusates with URC and UR. In conclusion, clinically available perfusate and hemodynamic parameters correlate well with donor characteristics and measured biomarkers in a discarded human NMP model.

MicroRNA-122–5p promotes renal fibrosis and injury in spontaneously hypertensive rats by targeting FOXO3

Hypertensive renal injury is accompanied by tubular interstitial fibrosis leading to increased risk for renal failure. This study aimed to explore the influences of miR-122–5p in hypertension-mediated renal fibrosis and damage. 14-week-old male SHR and WKY rats were randomly assigned to treat with rAAV-miR-122–5p or rAAV-GFP for 8 weeks. There were marked increases in miR-122–5p and Kim-1 levels and decreases in FOXO3 and SIRT6 levels in hypertensive rats. Transfection with rAAV-miR-122–5p triggered exacerbation of renal fibrosis, apoptosis and inflammatory injury in SHR, associated with downregulated levels of FOXO3, SIRT6, ATG5 and BNIP3 as well as upregulated expression of Kim-1, NOX4, CTGF, and TGF-β1. In cultured primary mouse renal tubular interstitial fibroblasts, exposure to angiotensin II resulted in obvious downregulation of FOXO3, SIRT6, ATG5, BNIP3 and nitric oxide levels as well as augmented cellular migration, oxidative stress, and inflammation, which were exacerbated by miR-122–5p mimic while rescued by miR-122–5p inhibitor and rhFOXO3, respectively. Notably, knockdown of FOXO3 strikingly blunted cellular protective effects of miR-122–5p inhibitor. In summary, miR-122–5p augments renal fibrosis, inflammatory and oxidant injury in hypertensive rats by suppressing the expression of FOXO3. Pharmacological inhibition of miR-122–5p has potential therapeutic significance for hypertensive renal injury and fibrosis-related kidney diseases.

Effect of Tamsulosin on Biomarkers after Kidney Stones Lithotripsy

The disease of kidney stones is one of the urinary system’s oldest and most common issues. Children with kidney stones have also become more popular, In recent years. Kidney stones are also very painful, and in some people, they can continue to occur. Kidney stone attacks result in over 2 million visits to health care facilities . People tend to get stones in the middle of life. During midlife, family and work commitments are at their height, which makes kidney stones costly. The diagnosis, treatment and prevention of kidney stones and the time away from work due to stones cost approximately $5.3 billion a year. To understand the effect renal some biomarkers in patients with kidney stones, before and after Lithotripsy and to evaluate the effect of Tamsulosin on these biomarkers in patients. Forty patients and twenty healthy persons were participate in this case-control study, Bl.urea, level of ALP, Level of KIM1, and level of Na c were investigate in serum with kidney stones patients and control healthy persons. The results were shown significant elevation in the levels of KIM-1, and serum urea were decreased significantly (p value &lt;0.05) while the levels of ALP and serum sodium show an improvement when compared to patients who were not used tamsulosin but it’s results were statistically not significant. the results were given an evidence for the crucial role of some biomarker in serum in patients with kidney stones, also the effects of using tamsulosin for 14 days in patients with kidney stones who underwent lithotripsy compared with those patients who were not used tamsulosin.

Choline and citicoline ameliorate oxidative stress in acute kidney injury in rats.

The purpose of this study is to investigate the effects of cholinergic anti-inflammatory pathway (CAP)-activating drugs, choline and citicoline (Cytidinediphosphate-choline, CDP-choline), on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) parameters and the contribution of NADPH Oxidase4 (NOX4) p22phox. Endotoxemia induces a systemic inflammatory response characterized by the production of pro-inflammatory mediators and reactive oxygen species (ROS), which eventually develops acute kidney injury (AKI). NADPH Oxidase4 (NOX4) p22phox pathway contributes to the development of endotoxemia-induced AKI. Inflammatory response can be controlled by CAP. Expressions levels of KIM-1, TNF-α, NOX4, p22phox and NFκB in the kidney tissues of rats were analyzed via RT-PCR in experimental groups; 1. Control, 2. LPS (10 mg/kg) + saline, 3. LPS + CDP-choline (375 mg/kg) and 4. LPS + choline (90 mg/kg). Choline and ROS levels in kidney tissues were also measured by a spectrofluorometric assay. LPS-induced elevations of ROS levels were decreased by CDP-choline or choline administration (p < 0.001). LPS-elevated KIM-1, TNFα, NOX4, p22 phox, and NFκB expressions were significantly decreased by choline or CDP-choline treatments (p < 0.001). Decreased ROS production in kidney tissues in treatment groups suggests that choline or CDP-choline may have therapeutic potential in endotoxemia-associated AKI via downregulating NOX4 and p22phox expressions (Tab. 1, Fig. 5, Ref. 45). Text in PDF www.elis.sk Keywords: endotoxemia, choline, cytidine diphosphate choline, acute kidney injury, reactive oxygen species.

Investigation of the nephroprotective effect of dexmedetomidine on colistin-induced nephrotoxicity in rats.

There are very few studies in the literature focusing on whether dexmedetomidine exerts a protective effect on colistin nephrotoxicity. Our study aims to investigate the nephroprotective effect of dexmedetomidine in an experimental model of nephrotoxicity in rats. The control group was administered saline (SF) intraperitoneally twice a day. The colistin group received an intraperitoneal (ip) injection of 10 mg/kg of colistin twice a day. The DX10 group received 10 mg/kg of colistin 20 minutes after the intraperitoneal injection of 10 mcg/kg of dexmedetomidine. The DX20 group received 10 mg/kg of colistin 20 minutes after the intraperitoneal injection of 20 mcg/kg of dexmedetomidine. Applications were continued for 7 days, twice a day. All rats were sacrificed on the 8th day after blood and kidney tissue samples were taken. BUN, Creatine, KIM-1 and Endothelin-1 were studied in blood samples. There was a significant difference in the median values of Urea, BUN and Creatine between the groups (p<0.001, p<0.001, p<0.001, respectively). There was a significant difference in the median values of KIM-1 and Endothelin-1 between the groups (p=0.009, p=0.001, respectively). A significant difference was observed between the histopathological scores of the groups (p<0.001). Dexmedetomidine significantly decreased the elevated levels of BUN, Creatinine, KIM-1, and Endothelin-1 induced by colistin. Dexmedetomidine, at both doses, histopathologically prevented apoptosis and reduced the number of necrotic cells in the kidneys. Dexmedetomidine provides renoprotective effects, therefore it is a valuable sedation agent for clinicians working in intensive care units (Tab. 2, Fig. 4, Ref. 19). Text in PDF www.elis.sk Keywords: rat, colistin, nephrotoxicity, dexmedetomidine.