Dilated Renal Tubules Research Articles

#1110 The AHR antagonist CH223191 alleviates kidney injury and renal fibrosis in adenine-induced chronic kidney disease

Abstract Background and Aims The aryl hydrocarbon receptor (AhR) is a transcription factor that can be activated by a variety of endogenous and exogenous ligands, and is involved in varieties of biological processes. It has been confirmed that the expression of AhR in kidney is elevated in chronic kidney disease (CKD). However, the specific role of AhR signaling pathway in CKD and its molecular mechanism remain unclear. This study aims to investigate the role and underlying mechanism of AhR in CKD, providing potential targets for the treatment of CKD. Method Male C57/B6L mice were randomly divided into control group, model group and treatment group. The model group was fed 0.2% adenine diet for 14 days to establish CKD model, while the treatment group was treated with AhR antagonist CH223191. AhR agonist kynurenine and antagonist CH223191 was applied in mouse tubular epithelial (TCMK-1) cells to investigate the underlying mechanisms of AhR in vitro. The renal function, histological changes and renal fibrosis in mice were evaluated to explore the role of AhR in adenine-induced CKD. Histological changes were evaluated by PAS and MASSON stainings, immunohistochemistry and immunofluorescence. Western blot and qPCR were used to measure protein/gene transcription levels. Results The levels of serum creatinine (p < 0.0001) and urea nitrogen (p < 0.0001) in the model group were significantly increased, with obvious renal tubule dilation and renal fibrosis by PAS and MASSON staining. CH223191 treatment significantly reduced serum creatinine (p=0.0012) and urea nitrogen ( < 0.0001) levels in mice, and alleviated renal fibrosis histologically. Q-PCR and Western-Blot results showed that the renal expression level of AhR in the treatment group was decreased, while the expression levels of fibronectin (Fn) (p < 0.0001), α-smooth muscle actin (α-SMA) (p < 0.0001), type I collagen (p=0.0007) and transforming growth factor-β (TGF-b) (p=0.0027) were significantly decreased. In addition, the expression levels of N-GAL (p < 0.0001) and KIM-1 ( < 0.0001) molecular markers associated with kidney injury were reduced, suggesting that inhibiting AhR expression in vivo can reduce adenine-mediated kidney injury and renal fibrosis in CKD. In vitro experiments indicated that kynurenic acid could induce the expression of AhR and AhR downstream regulatory gene CYP1b1 and upregulated fibrosis-related genes in tubular epithelial cells, while CH223191 treatment downregulated the fibrosis-related genes in tubular epithelial cells. Conclusion AhR mediates renal injury and renal fibrosis in adenine-induced CKD mice. AhR antagonist CH223191 exerts a renal protective effect by reducing the expression of AhR in renal tubular epithelial cells, thereby alleviating renal injury and renal fibrosis.

Functional metabolomics reveals arsenic-induced inhibition of linoleic acid metabolism in mice kidney in drinking water

Arsenic (As) is a heavy metal known for its detrimental effects on the kidneys, but the precise mechanisms underlying its toxicity remain unclear. In this study, we employed an integrated approach combining traditional toxicology methods with functional metabolomics to explore the nephrotoxicity induced by As in mice. Our findings demonstrated that after 28 days of exposure to sodium arsenite, blood urea nitrogen, serum creatinine levels were significantly increased, and pathological examination of the kidneys revealed dilation of renal tubules and glomerular injury. Additionally, uric acid, total cholesterol, and low-density lipoprotein cholesterol levels were significant increased while triglyceride level was decreased, resulting in renal insufficiency and lipid disorders. Subsequently, the kidney metabolomics analysis revealed that As exposure disrupted 24 differential metabolites, including 14 up-regulated and 10 down-regulated differential metabolites. Ten metabolic pathways including linoleic acid and glycerophospholipid metabolism were significantly enriched. Then, 80 metabolic targets and 168 predicted targets were identified using metabolite network pharmacology analysis. Of particular importance, potential toxicity targets, such as glycine amidinotransferase, mitochondrial (GATM), and nitric oxide synthase, and endothelial (NOS3), were prioritized through the “metabolite-target-pathway” network. Receiver operating characteristics curve and molecular docking analyses suggested that 1-palmitoyl-2-myristoyl-sn-glycero-3-PC, linoleic acid, and L-hydroxyarginine might be functional metabolites associated with GATM and NOS3. Moreover, targeted verification result showed that the level of linoleic acid in As group was 0.4951 μg/mL, which was significantly decreased compared with the control group. And in vivo and in vitro protein expression experiments confirmed that As exposure inhibited the expression of GATM and NOS3. In conclusion, these results suggest that As-induced renal injury may be associated with the inhibition of linoleic acid metabolism through the down-regulation of GATM and NOS3, resulting in decreased levels of linoleic acid, 1-palmitoyl-2-myristoyl-sn-glycero-3-PC, and L-hydroxyarginine metabolites.

<i>Curcuma longa</i> renal historestorative effects on sildenafil induced nephrotoxicity among male albino rats

Background: Sildenafil is a phosphodiesterase type 5 inhibitor mostly used in management of erectile dysfunction and pulmonary hypertension. On the other hand, Curcuma longa is herbal plant that is mostly used in diets among the Asian and African population and commonly used in treatment of respiratory, renal and dermatological diseases.
 Methods: A post-test only true experimental study design was utilized with 25 Albino rats grouped as follows: negative control, positive control (sildenafil 1µg/g bwt), low dose Curcuma longa (38.75mg/kg), medium dose Curcuma longa (77.5mg/kg) and high dose Curcuma longa (155mg/kg) each having 5 rats. Induction of nephrotoxicity using sildenafil was done for 15 days with an interim sacrifice for negative and positive control groups done on day 15. Experimental groups were sacrificed on day 22 after receiving Curcuma longa at respective doses. Blood samples for renal function tests were obtained on day 1, 15 and 22. Enblock harvesting of the kidneys after euthanizing the rats with concentrated CO2 and 10% neutral buffered formalin perfusion was done on day 15 and 22 then fixed in 10% neutral buffered formalin for 24hours. The kidneys were then processed for Hematoxylin and Eosin staining and photomicrographs were taken using Olympus light microscope fitted with LABOMED ivu 3100 digital camera.
 Results: In sildenafil induced nephrotoxicity group; glomerulus was distorted, dilated bowman`s space, dilated renal tubules and necrotic epithelial cells. Minimal histological changes were observed in low Curcuma longa dose group while in medium and high Curcuma longa dose groups; the glomerulus had well defined margins, bowman`s space reduced in size, epithelial cells appeared normal.
 Conclusion: It can be concluded that medium and high dose Curcuma longa have kidney historestorative effects in sildenafil induced nephrotoxicity among male albino rats.

The Effect of Different Concentrations of Bee Venom on Kidney Tissue of Albino Male Rats Rattus

In this study, 60 males of albino rats (Rattus rattus) were tested to study the effect of BV on kidney tissue at two doses of 10 and 40 µg/kg and two times after 10 and 15 days. The sample included 40 rats divided into two groups; 20 rats for each group. These groups were injected intraperitoneally according to the mentioned periods and were considered as a control group. The results exhibited renal tubule dilation and slight atrophy of glomerular tuft in the kidneys of rats treated with 10 µg/kg for 10 days compared to kidneys in the control treatment. Furthermore, 10 µg/kg treatment for 15 days has caused renal tubule dilation and degeneration with cytoplasmic vacuolation. Our findings also showed a slight atrophy of the glomerular tuft as well as a reduction in the number of glomeruli in this subcapsular area. The results also indicate regeneration of kidney tubules in rats given 40 µg/kg for 10 days. Meanwhile, sections of the kidney for rats that were given 40 µg/kg for 15 days exhibited a slight atrophy of glomerular tuft hypertrophied tubular epithelial cells.

Comparative valuation of the chlorpyrifos, acetamiprid, and lambda-cyhalothrin toxicity and their hematological and histopathological consequences in pigeons.

Globally agrochemicals are widely used in the agricultural sectors, posing potential eco-toxicological risks and disrupting various lifeforms including birds. Thus, the current work was conducted to compare the acute toxic impacts of pesticides (e.g., chlorpyrifos, acetamiprid, and lambda-cyhalothrin) on the pigeon's health. In total 50 adult pigeons were purchased from a local market where these pigeons were fed on pollution-free food. Post adaptation period (15 days), the pigeons were arbitrarily separated into five distinct groups after having been identified in this manner by chance (each group containing 10 pigeons). Control group (group 1) was not treated with any pesticide while the remaining groups (groups 2, 3, and 4) were treated with 0.25-mg/kg body weight of chlorpyrifos, acetamiprid, lambda-cyhalothrin, and a mixture of all three pesticides (group 5), respectively. After 36 days of exposure, the groups that had been exposed to the pesticide showed a significant (p < 0.05) increase in both the total number of platelets and the number of white blood cells (WBCs), in comparison to the control group. On the other hand, the groups that were exposed to the insecticides had significantly lower levels of red blood cells (RBCs), hemoglobin (Hb), and packed cell volume (PCV) (p < 0.05). The value of mean corpuscular volume (MCV) was significantly (p < 0.05) reduced in acetamiprid-exposed group, while a significant increase was observed in other pesticide-exposed groups. Obvious histopathological changes were observed in the tissues of control group and no such changes were reported by control group. Necrosis, pyknosis, lymphocyte infiltration, congestion of blood, dissolution of plasma membrane, and vacuolation were observed in the livers of pesticide-treated pigeons. The intestinal study showed the formation of goblet cells, villi rupturing, degeneration of serosa, necrosis, and pyknosis in treated groups. Renal alterations, dilation of renal tubules, reduction of glomerulus tissue, and edema were observed. This study manifests that the uncontrolled use of pesticides impairs ecosystems and poses a substantial health risk to wildlife and ultimately to human.

Cystic Clear Cell Renal Cell Carcinoma: A Morphological and Molecular Reappraisal.

A wide variety of renal neoplasms can have cystic areas. These can occur for different reasons: some tumors have an intrinsic cystic architecture, while others exhibit pseudocystic degeneration of necrotic foci or they have cystically dilated renal tubules constrained by stromal neoplastic cells. Clear cell renal cell carcinoma (CCRCC), either solid or cystic, is the most frequent type of renal cancer. While pseudocysts are found in high-grade aggressive CCRCC, cystic growth is associated with low-grade indolent cases. The latter also form through a cyst-dependent molecular pathway, and they are more frequent in patients suffering from VHL disease. The differential diagnosis of multilocular cystic renal neoplasm of low malignant potential and clear cell papillary renal cell tumor can be especially hard and requires a focused macroscopical and microscopical pathological analysis. As every class of renal tumor includes cystic forms, knowledge of the criteria required for a differential diagnosis is mandatory.

Zinc chelator treatment in crush syndrome model mice attenuates ischemia–reperfusion-induced muscle injury due to suppressing of neutrophil infiltration

In crush syndrome, massive muscle breakdown resulting from ischemia–reperfusion muscle injury can be a life-threatening condition that requires urgent treatment. Blood reperfusion into the ischemic muscle triggers an immediate inflammatory response, and neutrophils are the first to infiltrate and exacerbate the muscle damage. Since free zinc ion play a critical role in the immune system and the function of neutrophils is impaired by zinc depletion, we hypothesized that the administration of a zinc chelator would be effective for suppressing the inflammatory reaction at the site of ischemia–reperfusion injury and for improving of the pathology of crush syndrome. A crush syndrome model was created by using a rubber tourniquet to compress the bilateral hind limbs of mice at 8 weeks. A zinc chelator N,N,N′,N′-tetrakis-(2-pyridylmethyl)-ethylenediamine (TPEN) was administered immediately after reperfusion in order to assess the anti-inflammatory effect of the chelator for neutrophils. Histopathological evaluation showed significantly less muscle breakdown and fewer neutrophil infiltration in TPEN administration group compared with control group. In addition, the expression levels of inflammatory cytokine and chemokine such as IL-6, TNFα, CXCL1, CXCL2, CXCR2, CCL2 in ischemia–reperfusion injured muscle were significantly suppressed with TPEN treatment. Less dilatation of renal tubules in histological evaluation in renal tissue and significantly better survival rate were demonstrated in TPEN treatment for ischemia–reperfusion injury in crush syndrome. The findings of our study suggest that zinc chelators contributed to the resolution of exacerbation of the inflammatory response and attenuation of muscle breakdown in the acute phase after crush syndrome. In addition, our strategy of attenuation of the acute inflammatory reaction by zinc chelators may provide a promising therapeutic strategy not only for crush syndrome, but also for other diseases driven by inflammatory reactions.

Protective effect of Urtica dioica in liver and kidney damages induce by ethylene glycol in rabbits: A histopathological study

Urtica dioica is used in the many countries as seasoning and as an herbal medicine (antioxidant). In this study, adult male locale rabbit was separated into five groups, Group 1 considered as control, Group 2 to 5 animals were treated with 0.75% ethylene glycol (EG) in water to induce renal and liver damage till 30th day. The group 3 to 5 animals were feed extract of the plant of U. dioica at a dosage of 100 mg/kg body weight from day 15 to the day 30. The extracts were administered twice daily orally. Liver histopathological changes characterized by vacuolar degeneration and coagulative necrosis of hepatocytes, congestion and dilatation of central vein and sinusoids. Renal histopathological changes characterized by deposition of oxalates, blood vessels congestion infiltration of inflammatory cells and change in renal glomeruli in GE treated groups. While histopathological changes in the animals group treated with U. dioica extract and ethylene glycol showed an increase improvement of the histological features of liver tissue, were limited dilatation of renal tubules with less deposition of oxalates, as well as slight infiltration of inflammatory cells in the interstitial tissue. Finally, as a conclusion we noticed that U. dioica extracts had the ameliorative effect of ethylene glycol-induced hepatic and renal histopathological.

Therapeutic role of Azadirachta indica leaves ethanolic extract against diabetic nephropathy in rats neonatally induced by streptozotocin

ABSTRACT Diabetic nephropathy (DN) is manifested by chronic loss of renal function due to damage of glomeruli and renal tubules. Therefore, this study is mainly designed to evaluate the therapeutic role of Azadiracta indica (neem) leaves extract as a novel approach for treatment of DN in rats neonatally induced by streptozotocin (STZ). For this study, 40 offspring were selected after parturition and categorized into four groups (n = 10). Group1: control group, group 2: neem leaves extract supplemented group, group 3: diabetic group that injected with a single dose of STZ and group 4: diabetic group treated with neem extract. The results revealed deleterious histological and ultrstructural changes in the renal tissues of diabetic rats. Such changes included atrophied glomeruli, dilated renal cortical tubules and scattered hemorrhage spots, thickening of glomerular basement membrane, expansion of mesangial matrix and pyknotic podocyte. Additionally, the proximal convoluted tubule and distal tubule showed cytoplasmic vacuolation, vacuolated mitochondria, scattered lipid droplets, lost microvilli and disrupted basal lamina and basal infoldings. Moreover, significant decreased levels of serum antioxidants (SOD&CAT) and significant increased levels of serum MDA, urea and creatinine were noticed in diabetic rats. Neem leaves extract successfully alleviated the histological and ultrastructural as well as biochemical changes induced by diabetes.

​Safety Evaluation of Andrographolide-nanosuspensions

Background: Andrographolide (ANDRO) is a hydrophobic drug, which faces the problem of limited absorption due to poor water solubility. The current research prepared andrographolide nanosuspensions (ANDRO-NS) and examined in vivo toxicity for mice. Methods: ANDRO-NS were prepared by anti-solvent precipitation method, transmission electron micrographs, granularity analysis and in vitro release were used to characterize the ANDRO-NS, we evaluate the safety of the ANDRO-NS by using the acute toxicity test, local irritation test and chronic toxicity test. Result: The particle size of ANDRO-NS was (568.51±13.74 nm). The LD50 for ANDRO-NS was 548.91 mg/kg after oral administration to KM mice with a 95% CI of 468.19-645.03 mg/kg. The white blood cell counts and hemoglobin levels for the experimental groups were lower than controls receiving only saline. Serum aspartate transaminase, creatinine and blood urea nitrogen levels were greater than controls after 7 and 14 days of once-daily administration. After 14 days of administration, the platelet counts as well asalanine transaminase levels were, in addition, Histological observations indicated that interstitial kidney tissues were wider than controls and showed episodic bleeding after 7 days of administration. The highest dose administered also resulted in the dilation and blood engorgement of the central hepatic veins with some severed hepatic cords. Mice receiving the lowest dose of ANDRO-NS we administered appeared healthy and similar to controls receiving saline only. Following 14 days of administration, we found significant vacuolar degeneration of renal tubular epithelial cells and glomerular atrophy for the high-dose group as well as edema and necrosis in liver cells. The medium-dose group displayed kidney interstitial tissue widening with scattered bleeding, inflammatory cell infiltration and hepatocyte edema. The low-dose group displayed dilated renal tubules and irregularly-arranged liver cells as well as bleeding in the hepatic sinusoids. Therefore, short-term administration of andrographolide suspensions resulted in inflammation and time- and dose-dependent toxic effects on the kidneys and liver.

METTL3-mediated M6A methylation modification is involved in colistin-induced nephrotoxicity through apoptosis mediated by Keap1/Nrf2 signaling pathway

Colistin is a cationic polypeptide antibiotic. Despite its nephrotoxicity, it is still widely used as a last-line antibiotic against infection worldwide with the emergence of multi-drug resistant Gram-negative bacilli. N-methyladenosine (m6A) methylation-mediated degradation of RNA is essential for kidney development. However, m6A methylation impacts not only RNA stability, but also other RNA metabolism processes. How RNA decay affects the nephrotoxicity of colistin is largely unknown. Therefore, in this study, we verified that colistin could induce mouse kidney apoptosis through some apoptotic indicators, and confirmed the relationship between methylation and apoptosis through the detection of m6A methylation, thus elucidating the potential mechanism of colistin nephrotoxicity. The results showed that the renal tubule dilation and tubular structure were observed in the colistin group, and the oxidative stress index and ATPase activities were significantly different from those in the control group. Under electron microscope, the kidney in colistin group showed typical apoptotic morphological changes such as nuclear pyknosis, chromatin edge aggregation, and intact nuclear membrane, accompanied by significant changes in apoptosis-related genes. The level of m6A in the colistin group was significantly decreased, accompanied by downregulation of METTL3 mRNA and protein levels, and METTL3 was significantly correlated with apoptotic gene proteins. Data from this study suggested that m6A methylation was involved in oxidative stress-mediated apoptosis in the mechanism of colistin nephrotoxicity.

Investigations on Blood Physiology, Tissues Histology and Gene Expression Profile of Fusarium oxysporum Challenged Fish

Labeo rohita was challenged with Fusarium oxysporum to demonstrate a thorough immune response. Significant alterations in hematological parameters viz; erythrocyte, hemoglobin, hematocrit, mean corpuscular hemoglobin concentration, mean corpuscular volume, packed cell volume, platelets, and leukocytes were observed. Total serum protein, albumin, globulin, A/G ratio, aspartate aminotransferase, alkaline aminotransferase, alkaline phosphatase activity, and cholesterol were also altered in the infected fish. Marked histopathological lesions in gills, liver, and kidney (fusion of gill lamella, necrosis, edema, vacuolization, glomerulus congestion, dilated bowman's space and renal tubules alterations) were recorded. Tissue specific gene expression data showed that interleukin was upregulated in infected fish gills, liver, and kidney but was downregulated in blood. Tumor necrosis factor-α expression was upregulated in blood, liver, and kidney and was downregulated in gills. Upregulation of interferon-γ was observed in blood, gills, and kidney but downregulated in liver. All the cellular and molecular findings from this study can be taken as critical biomarkers in the advancement of diagnostic techniques against fungal infections as a part of wide-ranging programs of systematic and comprehensive monitoring of fish health and immune system.

A Multitubular Kidney-on-Chip to Decipher Pathophysiological Mechanisms in Renal Cystic Diseases.

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is a major renal pathology provoked by the deletion of PKD1 or PKD2 genes leading to local renal tubule dilation followed by the formation of numerous cysts, ending up with renal failure in adulthood. In vivo, renal tubules are tightly packed, so that dilating tubules and expanding cysts may have mechanical influence on adjacent tubules. To decipher the role of this coupling between adjacent tubules, we developed a kidney-on-chip reproducing parallel networks of tightly packed tubes. This original microdevice is composed of cylindrical hollow tubes of physiological dimensions, parallel and closely packed with 100–200 μm spacing, embedded in a collagen I matrix. These multitubular systems were properly colonized by different types of renal cells with long-term survival, up to 2 months. While no significant tube dilation over time was observed with Madin-Darby Canine Kidney (MDCK) cells, wild-type mouse proximal tubule (PCT) cells, or with PCT Pkd1+/- cells (with only one functional Pkd1 allele), we observed a typical 1.5-fold increase in tube diameter with isogenic PCT Pkd1-/- cells, an ADPKD cellular model. This tube dilation was associated with an increased cell proliferation, as well as a decrease in F-actin stress fibers density along the tube axis. With this kidney-on-chip model, we also observed that for larger tube spacing, PCT Pkd1-/- tube deformations were not spatially correlated with adjacent tubes whereas for shorter spacing, tube deformations were increased between adjacent tubes. Our device reveals the interplay between tightly packed renal tubes, constituting a pioneering tool well-adapted to further study kidney pathophysiology.

Empagliflozin ameliorates symptoms of diabetes and renal tubular dysfunction in a rat model of diabetes with enlarged kidney (DEK).

BackgroundSodium-glucose cotransporter 2 (SGLT2) inhibitors are widely used to reduce hyperglycemia. The present study investigated the effects of a SGLT2 inhibitor, empagliflozin, on hyperglycemia in a novel rat model of non-obesity type 2 diabetes with enlarged kidney (DEK).MethodsMale DEK rats with non-fasting blood glucose concentrations ≤300 mg/dl and >300 mg/dl were classified as nondiabetic and diabetic, respectively. Groups of nondiabetic (control) and diabetic (DM-cont) rats were fed standard chow for 12 weeks, whereas another group of diabetic (DM-empa) rats was fed standard chow containing empagliflozin (300 mg/kg/day) for 12 weeks. Blood glucose, body weight, glucose tolerance, food and water intake, urinary volume, plasma and urinary biochemical parameters, and bone mineral density were measured, and their kidneys and pancreas histologically analyzed.ResultsTreatment with empagliflozin reduced blood glucose concentration and food intake in diabetic rats, but inhibited loss of adeps renis and led to body weight gain. Empagliflozin attenuated polyuria and polydipsia but increased plasma concentrations of total cholesterol, sodium and total protein toward normal level. Empagliflozin also significantly reduced urinary excretion of proteins and electrolytes and restored bone mineral density and plasma concentrations of valine and isoleucine to normal levels. Moreover, dilation of renal tubules and kidney enlargement were not attenuated in the DM-empa group.ConclusionThe response of DEK rats to empagliflozin differed from that of other diabetic animal models, suggesting that DEK rats have unique characters for studying and evaluating the multiple biological effects of SGLT2 inhibitors. These findings also indicted that empagliflozin could ameliorate systemic metabolism and improve renal tubule function in diabetic condition.

The role of PGC-1α and metabolic signaling pathway in kidney injury following chronic administration with 3-MCPD as a food processing contaminant.

3-Monochloropropane-1,2-diol (3-MCPD) as a byproduct of food processing and a carcinogenic agent has attracted much attention in the last decades. Kidney is the main target organ that is sensitive to the toxicity of 3-MCPD. Due to limited evidence about possible 3-MCPD toxicity, we design an investigation to determine the role of mitochondrial biogenesis following chronic oral administration of 3-MCPD (2, 4, 8 and 32mg/kg) for 2months in male C57 mice. The present study evaluated the affects of 3-MCPD in modulating metabolic signalling which is associated with Il-18, PGC-1α, Nrf-2 and Sir3 which are the major transcription factors. Our data confirms controversial behaviors after chronic exposure with 3-MCPD. Over expression of the PGC-1α and Sir3 and IL-18 were observed after exposure with 2,4 & 8mgkg-1 day-1 of 3-MCPD. In front, PGC-1α down-regulation occurs at the highest dose (32mg/kg) resulted in kidney injury. Based on the findings, PGC-1α plays an important role in the restoration of the mitochondrial function during the recovery from chronic kidney injury. We suggest that the PGC-1α can be consider as a therapeutic target in prevention and treatment of kidney injury after chronic exposure of 3-MCPD. PRACTICAL APPLICATIONS: 3-Monochloropropane-1, 2-diol (3-MCPD) existed in several foods, can induce nephrotoxicity, progressive nephropathy and renal tubule dilation following acute and chronic exposure. It revealed that 3-MCPD toxicity is related to metabolites which can cause oxidative stress and activation of cell death signaling. It seems that cytotoxicity of 3-MCPD has disruptive effect on kidney cells due to rise in ROS production and decrease in mitochondrial membrane permeability. These effects can lead to MPT pore opening, cytochrome c release and activation of programed cell death signaling pathway. Therefore, present study was investigated the role of PGC-1a and the metabolic signaling involved in 3-MCPD-induced nephrotoxicity for the first time. Our data revealed that up-regulation of mitochondrial biogenesis following chronic exposure with 3-MCPD accelerates recovery of mitochondrial and cellular function in kidney by deacetylation of histones, overexpression of transcription factors (PGC-1α, Nrf-2, and Sir3) and maintaining cellular homeostasis.

Protective Effects of Vitamin C on Ivermectin Induced Toxicity on Kidney Functions and Brain Tissue in Rabbits (Oryctolagus cuniculus

The aim of this study was to evaluate the effects of repeated intake of a high dose of Ivermectin (IVM) alone or with co-administration of Vitamin C on kidney biochemical parameters and histological structure of kidney and brain of male rabbits (Oryctolagus cuniculus). Male rabbits were distributed into four groups (5 rabbits/group) and treated for three consecutive weeks: receiving distilled water; IVM 2 mg/kg b.w. subcutaneously, 3 times a week; IVM + Vitamin C 20 mg/1mL by gavage and IVM + Vitamin C 200mg /kg of diet in food. IVM significantly (p < 0.05) increased the plasma level of creatinine compared to the control group. Co-treatment with vitamin C orally by gavage ameliorated this level compared to IVM group. The histopathological examination of kidney sections in IVM alone treated group showed dilated Bowmans’ spaces, dilated renal tubules, vascular congestion and cell infiltration. The brain tissues demonstrated vacuolated neuropil and degenerative changes in nerve cells in both the cerebral cortex and hippocampus of IVM-treated rabbits. However, these histological changes were moderate in both vitamin C co-treated groups. In conclusion, vitamin C has a protective effect against nephrotoxicity and neurotoxicity induced by subacute administration of a high dose of IVM.

POS-431 ALTERED REGULATION OF TRYPTOPHAN METABOLISM AND ARYL HYDROCARBON RECEPTOR DISTRIBUTION IN RODENT POLYCYSTIC KIDNEYS

Polycystic kidney disease (PKD) is an inherited disorder characterized by stimulated cell proliferation, increased fluid secretion into cysts, and interstitial inflammation in renal tissue. Many mechanisms of disease progression have been determined in both autosomal dominant and autosomal recessive PKD, but the exact signaling pathways responsible for cystogenesis remain unclear. To explore novel mechanisms of PKD progression, we evaluated altered metabolic activity and associated receptor expression and distribution in the kidneys of 4- and 20-week-old PCK rats, an orthologous model of human autosomal recessive PKD, by mass spectrometry, quantitative analysis, real-time RT-PCR and immunofluorescence. Based on mass spectrometry and quantitative analysis results, the abundances of kynurenine and kynurenic acid were significantly higher in both 4- and 20-week-old PCK kidneys compared with age-matched wild-type kidneys. Since kynurenine and kynurenic acid are endogenous agonists of aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor for activating metabolic enzymes such as those in the cytochrome P450 (CYP) superfamily, the expression of AhR was detected by real-time RT-PCR in the kidneys of wild-type and PCK rats. At 4-weeks of age with early stage PKD, the expression levels of AhR were significantly lower in PKD compared with wild-type kidneys. Intriguingly, its expression level in 20-week-old PCK kidneys with progressive stage PKD was higher compared with wild-type kidneys. Since cysts are derived from medulla in PCK kidneys, the distribution of AhR was observed by immunofluorescence mainly in renal medullary specimens. AhR is known to translocate into the nucleus upon ligand binding, and in normal kidneys at 4 weeks of age, AhR was localized to the nucleus in tubular epithelia, but not at 20 weeks of age in normal kidneys. However, in PCK kidneys at 20 weeks of age, AhR was localized in the nucleus in normal tubular epithelia and in developed cysts, while in dilated renal tubules (which develop into cysts), AhR translocated to the nucleus in PCK kidneys both at 4 and 20 weeks of age. Indolamine-2,3-dioxygenase 2 (IDO2) is an enzyme which catalyzes the conversion from tryptophan to kynurenine/kynurenic acid. In epithelial cells of dilated tubules in PCK kidneys, IDO2 was localized in the cytoplasm, suggesting that IDO2 may have a role in activating the AhR signaling pathway in cystogenesis. The current findings suggest that the progression of PKD may relate to the increased activation of tryptophan metabolism in combination with altered expression and distribution of AhR. Regulation of the kynurenine/kynurenic acid and AhR signaling pathway may have therapeutic potential against disease progression in PKD.

Reduced Cleavage of Von Willebrand Factor By ADAMTS13 Exacerbates Acute Kidney Injury Secondary to Traumatic Injury

Introduction: Increases in plasma von Willebrand Factor (vWF) antigen levels, accompanied by decreases in its respective metalloprotease, ADAMTS13, have been demonstrated following severe injury. We hypothesized that following traumatic injury, endothelial damage and activation would result in the release of high molecular weight multimeric (HMWM) vWF forms into circulation with a concominant decrease in ADAMTS13. We further hypothesized that the alterations in circulating levels and functions of these factors, following traumatic injury, promote an increased thrombogenic and inflammatory response, which exacerbates the risk of developing post traumatic acute kidney injury (AKI). Methods: Plasma samples from a cohort of patients with trauma induced AKI, collected at 0-, 24-, and 72-hours after admission, were evaluated against plasma samples from trauma patients that were matched for injury characteristics, but were without AKI. VWF and ADAMTS13 activity and antigen levels were evaluated. Circulating vWF multimeric composition was determined by vertical agarose gel electrophoresis. We further evaluated the effects of traumatic injury on ADAMTS13 and vWF by means of a pre-clinical model of trauma and hemorrhage, which is comprised of a liver laceration, cardiac puncture, and soft tissue injury. Following this polytrauma model, hepatic ADAMTS13 RNA expression was evaluated by qRT-PCR of total liver RNA; analyzed liver samples were taken immediately before, 24-, and 72-hours after injury. The impact of the polytrauma model on renal function was evaluated via changes in plasma cystatin C, while the effect on the renal microvasculature was evaluated by means of immunofluorescent staining of vWF and fibrinogen in kidney sections. Results: VWF antigen was increased in trauma patients when compared to healthy controls upon admission (11.0 ± 3.72 ug/mL vs. 4.64 ± 0.91 ug/mL), persisting through 72-hours (13.11 ± 3.16 ug/mL). The percentage of HMWM vWF forms in trauma patient samples was elevated at all time points compared to pooled normal plasma (PNP) (27.9 ± 4.5% and 15.9 ± 0.8). Trauma patient ADAMTS13 activity was decreased at all time points (0.48 ± 0.14 IU/mL vs. 1.27 ± 0.26 IU/mL). AKI patients showed a 1.54 ± 0.48-fold increase in vWF antigen levels over the 72-hour period (p=0.016), while there was no change in the control trauma group. Those who developed AKI demonstrated a larger proportion of HMWM vWF at 72-hours than their non-AKI counterparts (32.9 ± 2.4% vs 28.8 ± 3.2%). qRT-PCR of murine total liver RNA, following the polytrauma model, exhibited ADAMTS13 RNA expression levels that were 35% of normal at 24-hours and 60% of normal at 72 hours post-injury. A matched analysis of murine plasma cystatin C levels demonstrated a marked elevation 24-hours post polytrauma model (sham: 510ng/mL (435-561) vs trauma: 550 (479-701)). H&amp;E staining of kidney sections, which were harvested following the polytrauma model, demonstrated generalized renal tubule dilation and denudation, suggestive of post-traumatic acute tubular necrosis. Immunofluorescent studies of kidney sections from the same mice exhibited 8-15µm microvascular plugs that were positive for fibrinogen with a vWF core; renal immunofluorescent studies of control mice demonstrated no microvascular depositions. Conclusions: These data indicate that traumatic injury alters the circulating composition of ADAMTS13 and its target, vWF, shifting their equilibrium to one that promotes inflammation and thrombosis. In the immediate moments following injury, these mechanisms support life-saving hemostasis; however, as these changes extend into the following days, the early hemostatic benefit quickly shifts to a burden that may produce microvascular clot deposition, exacerbating pathologic states such as acute kidney injury. Disclosures No relevant conflicts of interest to declare.

The Protective Effect of Annona Extracts on Renal Histopathology Induced by Ethylene Glycol in Male Rabbits

Annona is one of the many plant extracts that have been explored owing to theiranti-inflammatory and anticancer effects. Male locale rabbit were separated into five groups, Group Iconsider as control, The 4th other groups were treated orally with 0.75% ethylene glycol (EG) to inducerenal damage this treatment done till 30th days. Group III to V animals were served as curative regimenand received flavonoids, glycosides and alkaloids extract of the plant of Annona at a dose of 100 mg/kgbody weight from 15th day to 30th day. The extracts was administered twice daily by oral route. significantkidney injury in EG animals as demonstrated by a significant elevation (P= 0.05) in serum activities urea,creatinine, total protein and calculated globulin levels. Meanwhile there was significant drop (P=0.05) inalbumin, and A/G ratio compared to the control and treated group , The levels of lipid peroxidation weresignificantly higher in the Serum of the groups of animals treated with EG , while the levels of SOD,CAT and GPx were significantly lower than animals were treatment of EG group with Annona significantlyelevated the antioxidant mechanisms differ with that before treatment. Renal Histopathological changesincludes deposition of oxalates in the renal tubules lumen, degeneration and necrosis of epithelium of renaltubules , congestion of blood cells , infiltration of inflammatory cells and atrophy in glomerular tuft leadingto expansion of bowmans spaces. While Histopathological changes in animals group treated with Annonaextract and ethylene glycol were limited dilatation of renal tubules with vascular changes with blood vesselscongestion and interstitial hemorrhage , reduced renal damage revealed by less degenerative and necroticlesion in tubular epithelium and no inflammatory reaction and the glomeruli appeared normal compared withethylene glycol sections.

SUN-457 ELDERLY ONSET RAPIDLY PROGRESSIVE RENAL DYSFUNCTION WITH KIDNEY ENLARGEMENT AND TUBULOINTERSTITIAL INJURY MIGHT BE A NEW DISEASE ENTITY OF CILIOPATHY

Autosomal dominant tubulointerstitial kidney disease (ADTKD) is a group of uncommon genetic disorders characterized by slowly progressive decline in kidney function and autosomal dominant inheritance. ADTKD is usually described as not showing enlarged kidneys. However, there have been several reports of rapidly progressive renal dysfunction in elderly patients with bilateral kidney enlargement and ADTKD-like histology in Japan. Here we reported three cases with enlarged kidneys who have undergone dialysis with rapid progression and performed analysis of gene panel for target sequence of inherited kidney diseases. CASE PRESENTATION: 【Cases 1】 Eighty-four-year-old Japanese woman without family history of kidney diseases was noted to have renal dysfunction of unknown etiology 1 year prior to admission. She had developed progressive renal dysfunction (Cr 3.7mg/dL). Her abdominal CT revealed enlarged kidneys and histological findings showed marked tubular dilatation with extensive fibrosis in the interstitium, consistent with ADTKD. She was initiated hemodialysis 3 month after. 【Cases 2】 Seventy-four-year-old woman suffering from ulcerative colitis was noted to have mild renal impairment (Cr 1.1mg/dL) 1 year prior to admission. She developed progressive renal dysfunction (Cr 3.6mg/dL). Her serial abdominal CT showed enlarged kidneys on admission, but the kidneys were not enlarged one year ago. Her histological findings revealed consistent with ADTKD. She was initiated hemodialysis 5 month after. 【Cases 3】 Eighty-year-old woman was noted to have renal dysfunction (Cr 1.9mg/dL) 1 year prior to admission. She had developed progressive renal dysfunction (Cr 3.3mg/dL). Her CT showed enlarged kidneys and histological findings revealed consistent with ADTKD. She was initiated hemodialysis 1 year after. DISCUSSIONS: We searched the related literature with similar characteristics such as elder and rapid progression, enlarged kidneys, dilated tubules. And we found 15 cases which were only reported in Japan and compared among them including our 3 cases. Most cases are sporadic and seem to be non-inherited. The age at renal biopsy was 70 years. Before renal biopsy, a rate of Cr elevation was 0.5 mg / dL / month, and renal biopsy was performed at 4.1 mg / dL of Cr. Urine protein were positive in all cases, urine occult blood was positive in 50% cases. Urinary beta 2 microglobulin was 12145 ng/ml. Kidney enlargement was described in all cases except 2 undescribed cases. Renal biopsy findings showed minor glomerular changes and marked dilated renal tubules and interstitial fibrosis. The dilated tubules were detected as distal tubules by immunohistochemical staining. Electron microscopy showed irregularities, thinning and partial thickening of the tubular basement membrane. These pathological findings were compatible with traditional ADTKD. After renal biopsy, all cases showed rapidly progressive renal dysfunction with a rate of Cr elevation at 1.2 mg / dL / month and initiated dialysis in 4.1 months. Analysis of gene panel for target sequence of inherited kidney diseases was performed in 6 cases and no known gene mutation (including ADTKD-MUC1and ADTKD-UMOD) was detected. Elderly onset rapidly progressive renal dysfunction with enlarged kidneys and tubulo-interstitial injury might be a new disease entity of ciliopathy unlike traditional ADTKD.