Untreated Kidney Research Articles

A Qualitative Study to Assess the Living Experience of the Patient with Chronic Kidney Disease Undergoing Hemodialysis at Sharda Hospital, Greater Noida, Uttar Pradesh

Chronic kidney disease is a major problem in the world. CKD patients are increasing day by day because chronic kidney disease affecting the both the countries developed or developing county. Millions people are dying due chronic kidney disease. In developing countries, treatment with hemodialysis or kidney transplantation creates a hug financial burden for the majority of the people who need it. In another 112 countries, many people cannot afford treatment at all, resulting in the death of over millions people annually from untreated kidney failure.

Case Report : Hepatic Cirrhosis and Nephrolithiasis

Liver cirrhosis and nephrolithiasis are significant health problems with different pathophysiological mechanisms. Liver cirrhosis, the final stage of chronic liver disease, is characterized by fibrosis and regenerative nodules, leading to severe complications, including portal hypertension and liver failure. Nephrolithiasis, commonly known as kidney stones, is a common urological condition characterized by the formation of stones in the renal system. Although these conditions are distinct, metabolic and physiological changes in cirrhotic patients may predispose to nephrolithiasis. It was reported that a man came to the emergency room of RSUD Dr. Suhatman MARS with complaints of left-sided low back pain that had been felt for 1 week. The complaint was accompanied by shortness of breath which was getting worse over time. The patient also felt cough, headache, nausea, fatigue, urine slightly coloured like tea, weight loss. The patient has a history of untreated kidney stones. The patient has a history of hepatitis 31 years ago. The main objective of investigating the coexistence of liver cirrhosis and nephrolithiasis is to understand the relationship between these two conditions and to develop effective management strategies that can address their combined impact on patients.

Visualization of renal rotenone accumulation after oral administration and in situ detection of kidney injury biomarkers via MALDI mass spectrometry imaging.

The examination of drug accumulation within complex biological systems offers valuable insights into the molecular aspects of drug metabolism and toxicity. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) is an innovative methodology that enables the spatial visualization and quantification of biomolecules as well as drug and its metabolites in complex biological system. Hence, this method provides valuable insights into the metabolic profile and any molecular changes that may occur as a result of drug treatment. The renal system is particularly vulnerable to adverse effects of drug-induced harm and toxicity. In this study, MALDI MSI was utilized to examine the spatial distribution of drug and renal metabolites within kidney tissues subsequent to a single oral dosage of the anticancer compound rotenone. The integration of ion mobility spectrometry with MALDI MSI enhanced the data acquisition and analysis, resulting to improved mass resolution. Subsequently, the MS/MS fragment ions of rotenone reference drug were detected and characterized using MALDI HDMS/MS imaging. Notably, drug accumulation was observed in the cortical region of the representative kidney tissue sections treated with rotenone. The histological examination of treated kidney tissues did not reveal any observable changes. Differential ion intensity of renal endogenous metabolites was observed between untreated and rotenone-treated tissues. In the context of treated kidney tissues, the ion intensity level of sphingomyelin (D18:1/16:0), a sphingolipid indicator of glomerular cell injury and renal damage, was found to be elevated significantly compared to untreated kidney tissues. Conversely, the ion intensities of choline, glycero-3-phosphocholine (GPC), inosine, and a lysophosphatidylcholine LysoPC(18:0) exhibited a significant decrease. The results of this study demonstrate the potential of MALDI MSI as a novel technique for investigating the in situ spatial distribution of drugs and renal endogenous molecules while preserving the anatomical integrity of the kidney tissue. This technique can be used to study drug-induced metabolism and toxicity in a dynamic manner.

Investigation of reno-protective efficacy of thymoquinone in a unilateral hydronephrosis model

ABSTRACT We aimed to evaluate the effects of the antioxidant thymoquinone on treated and untreated kidneys on histological and oxidative parameters as well as Kidney Injury Molecule (KIM-1) levels in an experimental unilateral ureteropelvic junction obstruction (UPJO) with resultant hydronephrosis (HN) model. In adherence to the Animal research: reporting of in vivo exepriments guidelines, 34 male Wistar rats were randomly divided into four groups which were named accordingly: “CO” (corn oil), “TQ” (thymoquinone and corn oil), “HNCO” (UPJO-HN and corn oil), “HNTQ” (UPJO-HN, thymoquinone and corn oil). Histologically, pelvic epithelial damage, glomerular shrinkage and sclerosis, tubular damage, interstitial edema-inflammation-fibrosis (IEIF), and vascular congestion were assessed. Biochemically, malondialdehyde (MDA), superoxide dismutase (SOD), glutathione reductase (GR) and KIM-1 levels were assessed. Macroscopic HN developed in all obstructed kidneys. Ipsilateral obstructed kidneys deteriorated in all histological parameters. Thymoquinone attenuated glomerular shrinkage and sclerosis alterations but increased vascular congestion. Contralateral non-obstructed kidneys also showed histological deterioration. Thymoquinone had beneficial effects in terms of IEIF presence in contralateral kidneys but it increased vascular congestion. MDA and SOD results were inconclusive. UPJO caused decreased GR levels in the ipsilateral kidneys but not in the contralateral ones. This effect was not ameliorated by thymoquinone treatment. KIM-1 levels were increased in ipsilateral obstructed kidneys with a lower level in HNTQ group than in HNCO. KIM-1 level of the ipsilateral HNTQ group was higher than in both non-obstructed ipsilateral kidney groups. The effect of thymoquinone in ameliorating bilaterally observed histological alterations was limited and controversial. Oxidative damage detected by GR measurements was not prevented by thymoquinone. Thymoquinone partially decreased the damage as evidenced by reduced KIM-1 levels in thymoquinone-treated obstructed kidneys.

#1548 The involvement of soluble fms-like tyrosine kinase-1 in the transition from acute kidney injury to chronic kidney disease

Abstract Background and Aims Fms-like tyrosine kinase-1 (Flt-1) is a receptor for vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) and, playing a crucial role in angiogenesis and inflammation pathways. Soluble Flt-1 (sFlt-1) acts as a decoy for both VEGF and PlGF, regulating Flt-1-mediated angiogenesis and inflammation. We investigated the involvement of sFlt-1 in the transition from acute kidney injury to chronic kidney disease (CKD). Method Wild-type (WT) and sFlt-1 knock-out (KO) mice underwent ischemia-reperfusion injury (IRI) treatment. A unilateral 23-minute IRI and contralateral nephrectomy were performed to examine kidney function (AKI to CKD) for 2 weeks. To explore the downstream signaling pathway for sFlt-1 in the kidney, a unilateral 28-minute IRI was performed, and angiogenesis and cytokine expression arrays (Proteome Profiler Array Kits) were used to compare the relative expressions between 28-minute IRI and the contralateral untreated kidney cortices in both WT and sFlt-1 KO mice. Results A plasma sFlt-1 concentration was elevated just after IRI and reached its peak after 12 hours. Blood urea nitrogen (BUN) and plasma creatinine (pCre) levels peaked at 24 hours after IRI. The peak levels of BUN and pCre were significantly lower in sFlt-1 KO compared to those in WT mice (BUN 43.23 ± 5.9 mg/dL and 82.7 ± 13.0 mg/dL, p = 0.008, pCre 0.18 ± 0.03 mg/dL and 0.31 ± 0.04 mg/dL, p = 0.008 in KO and WT mice, respectively). Urinary KIM-1 (uKIM-1) levels 12 hours after IRI in WT mice were higher than those in sFlt-1 KO mice (WT vs sFlt-1 KO; uKIM-1 4.82 ± 0.83 pg/day vs 2.39 ± 0.98 pg/day, p = 0.09). Histopathological evaluation of the kidneys showed reduced monocyte infiltration in sFlt-1 KO mice compared to WT mice 14 days after IRI. In cytokine expression assay of kidney cortices 3 days after IRI, IRI-induced increased expressions of tissue repairs and immune response proteins such as TIMP-1, C5/C5a, and TREM-1 were alleviated in sFlt-1 KO mice compared to those in WT mice. In the Angiogenesis expression array of kidney cortices 1 day after IRI, there were no significant differences in the levels of IRI-induced increased expression of anti-/pro-angiogenesis factors, including TIMP-1, Cyr61, SerpinE1, angiogenin, platelet factor 4, collagen factor 3, MMP-9, and SDF-1, between WT and sFlt-1 KO mice. Conclusion The acute and chronic kidney damages after IRI was reduced in sFlt-1 KO mice compared to WT mice. The protective effect on the kidneys by eliminating sFlt-1 would be associated with immune response signaling such as TIMP-1, C5/C5a, and TREM-1 but not with anti-/pro-angiogenesis factors.

Successful treatment of an urgent dialysis patient with uremic encephalopathy

Diabetes has become a global pandemic and is believed to be one of the most common risk factors for chronic kidney disease, which leads to end-stage renal disease, of which a significant number will develop, requiring renal replacement therapy. Uremic encephalopathy is the most frequent complication of untreated chronic kidney disease, is defined as cerebral dysfunction due to toxin accumulation. If this occurs, renal replacement therapy, which is urgent dialysis, is essential for lowering the patient's mortality. We report a 48-year-old female patient with uncontrolled type 2 diabetes who presented with delirium, poor physical performance and muscle mass loss.

S-38-2: PREVENTION OF EXPERIMENTAL GENETIC HYPERTENSION - GENOMIC CLUES FROM THE KIDNEY

Brief treatment with renin-angiotensin system (RAS) inhibitors permanently reduces the genetic hypertension of the spontaneously hypertensive rat (SHR). The kidney is involved in this reprogramming, but relevant molecular genetic changes are unknown. We studied SHR kidney RNA differential expression (controlling for non-specific effects of lower BP) and DNA methylation 6 weeks after treatment with the angiotensin receptor blocker losartan from 10 to 14 weeks of age. RNA sequencing revealed a significant 6-fold (P < 0.0001) increase in renin gene (Ren) expression during treatment. Six weeks later direct mean arterial pressure remained lower than untreated SHR (123 vs 140 mmHg, P = 0.001) and kidney Ren expression was reduced by 23% (P = 0.03) and DNA methylation within the Ren promoter region was significantly increased (P = 0.04). Experiments with the ACE inhibitor perindopril and renin immunohistochemical analyses confirmed significant long-term reduction in kidney Ren expression following RAS inhibition. Genomic RNA sequencing analysis identified 13 candidate genes (Grhl1, Ammecr1l, Hs6st1, Nfil3, Fam221a, Lmo4, Adamts1, Cish, Hif3a, Bcl6, Rad54l2, Adap1, Dok4) and the miRNA miR-145-3p that were significantly differentially expressed and correlated closely with decreased Ren expression. Ten of the candidates were central to gene networks that exhibited significant enrichment for genes relevant to BP and the RAS. We propose that reduced renin gene expression is a legacy of early RAS inhibition and responsible for the persistent reduction in BP in SHR.

PS-B04-1: ANTI-HYPERTENSIVE AND ANTI-PROTEIURIC EFFECTS OF MELINJO SEED EXTRACT IN THE RAT REMNANT KIDNEY MODEL

Melinjo (Gnetum gnemon L.) is a fruit native to Indonesia and is considered as a healthy food. Melinjo resveratrol (MjR) is a resveratrol derivative extracted from the melinjo seed. Interestingly, recent literatures have documented that MjR exhibits the angiotensin converting enzyme inhibitory activity. Hence, we hypothesized that MjR exhibits beneficial effects in the chronic kidney disease. In order to test our hypothesis, MjR was administered in the rat remnant kidney (RK) model. Rats were divided into three groups: sham operated rats (SHAM, n = 4), untreated remnant kidney rats (RK, n = 5), RK rats treated with MjR 300 mg/kg/day (RK + MjR, n = 5). The rats were sacrificed at twelve weeks. Blood pressure and urinary protein levels significantly increased whereas creatinine clearance (CCr) significantly descread in the RK group compared to the SHAM group at twelve weeks. MjR significantly reduced systolic blood pressure (108 ± 2 vs. 147 ± 2 mmHg, p < 0.05) as measured by tail-cuff methods. Urine protein excretion was significantly ameliorated by MjR compared with that in RK (79.8 ± 54.8 vs. 269 ± 87.5 mg/day, p < 0.0015). Although Ccr was not altered in RK + MjR compared with RK rats, histological evaluation revealed that the glomerular sclerosis was attenuated in RK + MjR group. Podocytes in the RK rats showed increase in desmin, a marker of podocyte injury, which was alleviated with the administration of MjR. In conclusion, MjR reduced blood pressure, proteinuria and glomerular damage in the RK rats. These data suggest that MjR may mitigate the progression of chronic kidney disease, which needs further confirmation.

Osteitis Fibrosa Cystica; A Forgotten Manifestation of Secondary Hyperparathyroidism Due to End-Stage Renal Disease: A Case Report

IntroductionOsteitis fibrosa cystica is a rare complication of secondary hyperparathyroidism. Even though it is thought to be a disease of the past, it still continues to be seen in this modern era in the setting of undiagnosed or untreated chronic kidney disease.Case PresentationA 26-year-old married Ethiopian women presented with 4 year history of progressive proximal weakness of extremities and diffuse bone pain. Physical examination revealed diffuse bone tenderness and features consistent with myopathy. Blood work up showed raised creatinine, markedly elevated PTH, borderline low calcium, and vitamin D in severe deficiency range. X-Ray findings were consistent with classical skeletal lesion of severe secondary hyperparathyroidism.ConclusionOsteitis fibrosa cystica is a rare but still frequent complication of secondary hyperparathyroidism, which may be the initial presentation of chronic kidney disease. This case report emphasizes the importance of considering secondary hyperparathyroidism in patients presenting with weakness and bone pain to allow for early diagnosis, treatment, and improvement of overall prognosis.

L-Arginine’s Glucose Homeostatic Influence in Renal Damaged Wistar Rats is Possibly Mediated by Adiponectin Expression

Hypoglycemia, as a direct result of the glucosuria is usually seen in patients with acute kidney injury hence glucose homeostasis is disturbed. The compensatory effect of adiponectin in the insulin deficient state is documented. We previously demonstrated that L-arginine enhances glucose transport mechanisms in renal-damaged rats. The current study was designed to investigate possible glucose handling synergy between L-arginine and adiponectin in Wistar rats induced with acute kidney injury. Twenty four rats weighing between 120g-150g were divided into 4 groups of six rats each. Group 1 (Control) had normal feed and water; Group 2 (Untreated) was induced with AKI and left untreated. Group 3 and 4 took 50mg/kg and 500mg/kg L-arginine respectively after AKI induction. Acute kidney injury was induced by intra-muscular injection of glycerol (50% solution, 8 ml/kg BW). Oral glucose tolerance test, insulin response test, serum creatinine test and adiponectin assay (ELISA) were carried out. Data was analysed using one way ANOVA and expressed as mean± standard error of mean (SEM) with p ≤ 0.05 considered as significant. L-arginine induced rapid insulin-like action which was effective after thirty minutes of oral glucose loading when compared the control group. Glucose uptake from the blood was also more effective and quicker in the L-arginine treated groups. Adiponectin was significantly expressed across the test groups when compared to the control group and there was insulin resistance in the untreated kidney injured rats 1 hour into the insulin response test. It may be concluded that L-arginine has a homeostatic influence on glucose handling in kidney-damaged rats possibly mediated by increased adiponectin expression.

Characterization of pre-gelatinized kidney bean (Phaseolus vulgaris L.) produced using microwave hot-air flow rolling drying technique

This study aims to develop a pretreatment and drying process for kidney beans to improve their cooking efficiency. Hot air drying (HD) and microwave hot air rolling drying (MHRD) were analyzed for the drying of pre-cooked kidney beans, and the effect of hot air and microwave on structure and gelatinization properties of kidney beans starch were investigated. The result indicated that the drying time was significantly reduced by approximately 27% (p < 0.05) with the increase of hot air temperature by 10 °C in the HD-processed material. Based on the incorporation of microwave radiation, drying time was further decreased by 2.5 times at the same hot air temperature. The microstructure, crystalline structure, thermal behavior and FTIR spectra of the kidney bean starch suggested the positive correlation between the hot air temperature/microwave radiation and destruction of their crystalline structure/gelatinization degree. The cooked dried kidney beans exhibited significantly decreased hardness, gumminess and chewiness and increased springiness, compared with the untreated kidney beans, suggesting an improved cooking efficiency. Base on the results of drying efficiency, color profile, gelatinization of kidney bean starch and texture profile analysis of the cooked beans, MHRD-70 was the optimum drying process.

Life Table and Preference Choice of Frankliniella occidentalis (Thysanoptera: Thripidae) for Kidney Bean Plants Treated by Exogenous Calcium.

Simple SummaryWestern flower thrips, Frankliniella occidentalis, is an invasive key pest that damages vegetables and ornamentals worldwide. The activation of induced resistance by chemicals may provide a simple and feasible way of achieving improvement of resistance to stress in crop plants, which is an important technology for the development of sustainable agriculture. Calcium (Ca) is an essential element for plants; numerous studies have shown that Ca can confer crop plants with resistance to abiotic and biotic stresses. For the first time, we report the negative effects of exogenous Ca on kidney bean plants in relation to the performance of F. occidentalis, including a reduced preference of thrips. Therefore, Ca could potentially be used to control F. occidentalis.Exogenous calcium (Ca) has been used to induce host plant resistance in response to abiotic and biotic stresses, including from thrips attack. The aim of this study was to determine whether exogenously applied Ca affects the performance of Frankliniella occidentalis. We assessed the development time, total longevity, reproduction, and population parameters of F. occidentalis, and its preference choice on Ca-treated or untreated control kidney bean plants under laboratory conditions. The results showed that F. occidentalis fed on Ca-treated leaves had a longer developmental time but lower longevity (female and male) and fecundity than F. occidentalis fed on control leaves. Population parameters, including the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0), were all found higher in control leaves than in Ca-treated leaves, and the mean generation time (T) was shorter. In preference choices, the number of thrips on control plants was higher than the number of thrips on Ca-treated kidney bean plants. Overall, our results indicated that exogenous Ca pretreatment on kidney bean plants affected the life history and preference choice of F. occidentalis, suggesting Ca might be used as a promising elicitor of inducible plant defense against thrips.

Evaluation of polyherbal formulation against isoniazid and rifampicin induced oxidative stress in rat kidney

The present study deals with oxidative stress in rat kidneys upon administration of Isoniazid (INH) and Rifampicin (RIF), and the attenuative effect of polyherbal formulation Heptoplus. Heptoplus is a polyherbal drug that contains phyllanthin as the active ingredient. Rats were intoxicated with INH and RIF and Heptoplus was orally fed to rats along with INH and RIF. Body weight, serum biochemical, antioxidant status, mitochondrial enzyme activity, lysosomal enzyme leakage and the histopathological impact were studied in the rat kidneys. Body and kidney weight of untreated rats showed deviation, but on treatment, they maintained their normal weight. Untreated rats showed altered serum urea and creatinine levels with deviated antioxidant status and enzymes leakages from lysosomes and decreased mitochondrial enzyme activity in kindey. But Heptoplus treatment allowed the rats to maintain their biochemical indices in blood and kidney at near normal level. Histopathological reports revealed INH- and RIF -induced oxidative stress was not in sufficient amount to incur any significant damage at the tissue level. Hence, both treated and untreated rat kidneys exhibits normal architecture. In conclusion, Heptoplus treatment protects the kidney of rats from the INH- and RIF- induced oxidative stress.

Natural history of untreated kidney cancer

Natural history of untreated kidney cancer

Mir199a-3p and 5p Influence Fibrotic Disease States, And Increase In Expression After Radiation

The mir199a family, composed of 3p and 5p splice variants, are associated with fibrotic cardiac, lung and renal diseases of different etiologies. Mir199a-3p and 5p target vital negative regulators of fibrosis-inducing pathways including JAK/STAT via targeting of SOCS7, and caveolin-dependent internalization pathway through targeting of CAV1. The objective of this study was to investigate if mir199a 3p and 5p expression increase after irradiation (RT) and regulate radiation-induce fibrosis via the similar pathways. C57BL/6 were treated with focal 14Gy renal irradiation and kidneys and overlaying skin harvested after 24hrs, 10wks, or 20wks. Untreated mouse kidneys and skin served as age-matched controls. RNA was isolated from the skin and kidney tissue using a trizol extraction method, and quantitative PCR analysis was performed to assess gene expression changes in mir199a-3p and 5p and critical components of the JAK/STAT and caveolin pathways. Both mir199a-3p and 5p expression increased in kidneys at the 20wks, with increases over untreated controls of 2.1-fold (p<0.001) and 2.4-fold (p<0.001) respectively. A 10-fold increase in expression of mir199a precursor RNA was seen for skin at 20 wks, and this translated into a 3.1-fold increase of mir199a-3p (p = 0.046) but without a measurable increase in the mir199a-5p variant. SOCS7 expression levels decreased at 20wk consistent with mir199a targeting, although not significantly when comparing to untreated controls. Levels of CAV1 increased significantly over untreated controls at the 10wk (1.8-fold, p = 0.04). At 20 weeks, transcript levels of CAV1 decreased to a level no longer significantly different than in age-matched controls, consistent with mir199a targeting. We show that mir199a family members are elevated in the kidneys and overlaying skin after 14Gy of radiation and mechanistically-associated with fibrosis. On-going studies are evaluating mir199a-3p and 5p after more clinically-relevant doses of RT.

Consistent alteration of chain length-specific ceramides in human and mouse fibrotic kidneys

BackgroundSeveral studies revealed alterations of single sphingolipid species, such as chain length-specific ceramides, in plasma and serum of patients with kidney diseases. Here, we investigated whether such alterations occur in kidney tissue from patients and mice suffering from renal fibrosis, the common endpoint of chronic kidney diseases. MethodsHuman fibrotic kidney samples were collected from nephrectomy specimens with hydronephrosis and/or pyelonephritis. Healthy parts from tumor nephrectomies served as nonfibrotic controls. Mouse fibrotic kidney samples were collected from male C57BL/6J mice treated with an adenine-rich diet for 14 days or were subjected to 7 days of unilateral ureteral obstruction (UUO). Kidneys of untreated mice and contralateral kidneys (UUO) served as respective controls. Sphingolipid levels were detected by LC-MS/MS. Fibrotic markers were analyzed by TaqMan® analysis and immunohistology. ResultsVery long-chain ceramides Cer d18:1/24:0 and Cer d18:1/24:1 were significantly downregulated in both fibrotic human kidney cortex and fibrotic murine kidney compared to respective control samples. These effects correlate with upregulation of COL1α1, COL3α1 and αSMA expression in fibrotic human kidney cortex and fibrotic mouse kidney. ConclusionWe have shown that very long-chain ceramides Cer d18:1/24:0 and Cer d18:1/24:1 are consistently downregulated in fibrotic kidney samples from human and mouse. Our findings support the use of in vivo murine models as appropriate translational means to understand the involvement of ceramides in human kidney diseases. In addition, our study raises interesting questions about the possible manipulation of ceramide metabolism to prevent progression of fibrosis and the use of ceramides as potential biomarkers of chronic kidney disease.

COVID-19 and Kidney Disease Disparities in the United States

Racial, ethnic, socioeconomic, age, and sex-related health disparities in kidney disease are prominent in the United States. The Coronavirus Disease 2019 (COVID-19) pandemic has disproportionately affected marginalized populations. Older adults, people experiencing unstable housing, racial and ethnic minorities, and immigrants are potentially at increased risk for infection and severe complications from COVID-19. The direct and societal effects of the pandemic may increase risk of incident kidney disease and lead to worse outcomes for those with kidney disease. The rapid transition to telemedicine potentially limits access to care for older adults, immigrants, and people experiencing unstable housing. The economic impact of the pandemic has had a disproportionate effect on women, minorities, and immigrants, which may limit their ability to manage kidney disease and lead to complications or kidney disease progression. We describe the impact of COVID-19 on marginalized populations and highlight how the pandemic may exacerbate existing disparities in kidney disease.

P0029GLUCOSYLCERAMIDE SYNTHASE INHIBITION PRESERVES MITOCHONDRIAL FUNCTION AND REDUCES REACTIVE OXYGEN DAMAGE IN THE JCK MOUSE MODEL OF POLYCYSTIC KIDNEY DISEASE

Abstract Background and Aims Polycystic kidney disease (PKD) is characterized by the formation and progressive growth of renal cysts ultimately leading to end-stage renal disease (ESRD). Previous work has shown that treatment of PKD mouse models with a glucosylceramide synthase inhibitor (GCSi) inhibits glycosphingolipid metabolism and blocks disease progression. Clinical trials to determine the efficacy of GCSi treatment for ADPKD are currently ongoing. ADPKD patient kidneys, as well as kidneys from in vivo mouse models of PKD, display defects in mitochondrial morphology and function, leading to reactive oxygen species (ROS) generation. Functional defects include decreased fatty acid oxidation and increased glycolysis that can promote cellular proliferation. To determine if glucosylceramide synthase inhibition ameliorates mitochondrial dysfunction, we assessed the impact of GCSi treatment on mitochondrial formation and function in the jck mouse model of PKD. Method Twenty-six-day old WT or jck mice were treated with vehicle or 60 mg/kg GCSi (Genz667161) in feed for 38 days prior to tissue harvest. Kidney mRNA expression was measured using RT-PCR. Kidney protein levels were measured by western blot. Mitochondrial DNA content was measured using real-time PCR. Oxidized DNA was detected by immunohistochemistry using anti-8OHdG antibodies. Oxidized proteins were measured using the Oxyblot system (manufacturer). Results Reduced cystogenesis following GCSi treatment was associated with preserved mitochondrial number in jck mice as evidenced by increased levels of mitochondrial DNA, increased mitochondrial proteins (Tom20, VDAC, SirT3, and MT-Co1), and elevated mRNA expression (Nd3, Cox3, and Atp8). Stabilization of mitochondrial number was accompanied by activation of pathways that promote mitochondrial biogenesis, including increased PGC1α protein and Ppargc1a and Tfam1. Downregulation of several antioxidant genes (Gpx6, Gstk1, Prdx3, and Sod2) were observed in untreated jck samples. Consistent with this, levels of oxidized DNA and oxidized proteins were increased in untreated jck samples. Treatment with GCSi partially reversed the downregulation of antioxidant genes and decreased the levels of oxidized DNA and proteins to near WT levels in jck tissues. We then compared the jck model to tissues derived from ADPKD patients. Western blots demonstrate a reduction in electron transport chain (CxI-CxV) and mitochondrial outer-membrane markers in both untreated jck and ADPKD samples compared to controls. Similarly, the mitochondrial genes Nd1, Nd2, Cox2, and Atp6 were reduced in untreated jck kidneys and ADPKD samples relative to controls. As previously mentioned, we observed a generalized loss of antioxidant gene expression in untreated jck kidneys. Consistent with this, there were increased levels of both oxidized DNA and oxidized protein in untreated jck samples; this increase in DNA and protein oxidation was mirrored in ADPKD patient samples. Conclusion Mitochondrial number and function were reduced in both untreated jck mouse kidneys and human ADPKD samples. Increased oxidative stress, as evidenced by increased levels of oxidized DNA and proteins, is evident in both jck and ADPKD tissues. In vivo treatment with a GCSi effectively reversed the observed increase in oxidative stress and inhibited disease progression in the jck model of ADPKD. Our data demonstrate that reduced kidney cyst growth following GCSi treatment is associated with preserved mitochondrial formation and function.

Pharmacologic targeting of renal ischemia-reperfusion injury using a normothermic machine perfusion platform

Normothermic machine perfusion (NMP) is an emerging modality for kidney preservation prior to transplantation. NMP may allow directed pharmacomodulation of renal ischemia-reperfusion injury (IRI) without the need for systemic donor/recipient therapies. Three proven anti-IRI agents not in widespread clinical use, CD47-blocking antibody (αCD47Ab), soluble complement receptor 1 (sCR1), and recombinant thrombomodulin (rTM), were compared in a murine model of kidney IRI. The most effective agent was then utilized in a custom NMP circuit for the treatment of isolated porcine kidneys, ascertaining the impact of the drug on perfusion and IRI-related parameters. αCD47Ab conferred the greatest protection against IRI in mice after 24 hours. αCD47Ab was therefore chosen as the candidate agent for addition to the NMP circuit. CD47 receptor binding was demonstrated by immunofluorescence. Renal perfusion/flow improved with CD47 blockade, with a corresponding reduction in oxidative stress and histologic damage compared to untreated NMP kidneys. Tubular and glomerular functional parameters were not significantly impacted by αCD47Ab treatment during NMP. In a murine renal IRI model, αCD47Ab was confirmed as a superior anti-IRI agent compared to therapies targeting other pathways. NMP enabled effective, direct delivery of this drug to porcine kidneys, although further efficacy needs to be proven in the transplantation setting.

Effect of Mesenchymal Stem Cells Therapy on Cardiovascular Risk in Experimental Diabetic Kidney Disease

Abstract Background Bone Marrow-derived Mesenchymal stem cells (MSC)s were experimentally used in treating diabetes mellitus (DM) and its complications, particularly, diabetic kidney disease (DKD). Cardiovascular diseases are the leading cause of mortality in diabetic patients, which has been strongly associated with progression of DKD. Aim This study was designed to explore the effect of MSCs in modulating cardiovascular risk factors that develop with DKD. Study Design 60 adult female albino rats were allocated into three groups: control group, untreated diabetic kidney disease group (un-DKD) and MSCs treated diabetic kidney disease group (MSC-DKD). DM was induced by a single dose of Streptozotocin at a dose of 35mg/kg, then rats were sacrificed 8 weeks later. MSC-DKD rats were treated by IV injection of bone marrow-derived MSCs 4 weeks after DM induction. DKD establishment was confirmed by histopathological kidney changes and elevated plasma creatinine and urea in both un-DKD and MSC-DKD rats. Methods Blood Pressure and lipid profile were measured, and atherogenic index was calculated to assess cardiovascular risk accompanying the progression of the diabetic renal damage. Moreover, aortic reactivity studies were performed in vitro. Results A single MSCs injection to MSC-DKD rats resulted in significant decrease in final blood glucose, mean arterial blood pressure and atherogenic index, together with a significant increase in plasma HDL level compared to untreated rats, thus minimizing cardiovascular risk factors. MSC-DKD aortae also exhibited significant enhancement of vascular reactivity parameters, particularly vasorelaxation which could partially explain the improvement of blood pressure. On the other hand, DKD didn’t improve by MSCs therapy. These results conformed to tracking labelled MSCs which were found in abundance in both aortic and pancreatic tissues and absent in kidneys. Conclusion MSCs therapy holds hope of improving cardiovascular risk and mortality for diabetic patients with diabetic Kidney disease.