ATCC ® Normal Human Primary Renal Proximal Tubule Epithelial Cells, when grown in Renal Epithelial Cell Basal Media supplemented with Renal Epithelial Cell Growth Kit components, provide an ideal cell system to propagate renal epithelial cells in low serum (0.5% FBS) conditions. The cells are cryopreserved in the second passage to ensure the highest viability and plating efficiency. ATCC ® Primary Cell Solutions™ cells, media, supplements and reagents are quality tested together to guarantee optimum performance and reliability.
Cadmium (Cd2+) is a known nephrotoxin causing tubular necrosis during acute exposure and potentially contributing to renal failure in chronic long-term exposure. To investigate changes in global gene expression elicited by cadmium, an in-vitro exposure system was developed from cultures of human renal epithelial cells derived from cortical tissue obtained from nephrectomies. These cultures exhibit many of the qualities of proximal tubule cells. Using these cells, a study was performed to determine the cadmium-induced global gene expression changes after short-term (1 day, 9, 27, and 45 μM) and long-term cadmium exposure (13 days, 4.5, 9, and 27 μM). These studies revealed fundamental differences in the types of genes expressed during each of these time points. The obtained data was further analyzed using regression to identify cadmium toxicity responsive genes. Regression analysis showed 403 genes were induced and 522 genes were repressed by Cd2+ within 1 day, and 366 and 517 genes were induced and
Human Proximal Tubular Epithelial Cells from Creative Bioarray are isolated from normal human proximal tubular tissue. Human Proximal Tubular Epithelial Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based coating solution for 2 min and incubated in Creative Bioarrays Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells at passage 3 are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 0.5x10^6 cells per ml. Cells can be expanded for 3-7 passages at a split ratio of 1:2 or 1:3 under the cell culture conditions specified by Creative Bioarray. Repeated freezing and thawing of cells is not recommended ...
Renal Epithelial Cell Basal Medium is a sterile, phenol red-free, liquid tissue culture medium intended for use as one component in a complete ATCC ® Primary Cell Solutions ™ system. This system is designed to support epithelial cells (e.g., renal proximal tubule epithelial cells) derived from normal human kidney. Renal Epithelial Cell Basal Medium contains essential and non-essential amino acids, vitamins, other organic compounds, trace minerals and inorganic salts. To support the proliferation and plating efficiency of renal epithelial cells (e.g., Primary Renal Proximal Tubule Epithelial Cells, Normal, Human, ATCC ® PCS-400-010; Primary Renal Cortical Epithelial Cells, Normal, Human, ATCC ® PCS-400-011; Primary Renal Mixed Epithelial Cells, Normal, Human, ATCC ® PCS-400-012), Renal Cell Basal Medium must be supplemented with the Renal Epithelial Cell Growth Kit (ATCC ® PCS-400-040). Using Renal Epithelial Cell Basal Medium supplemented with the Renal Epithelial Cell Growth
Previous work from our laboratory indicates that the dopamine D2 receptor (D2R) in the kidney has a direct role in regulating renal inflammation and injury and blood pressure. Some common single nucleotide polymorphisms (D2R SNPs; rs 6276, 6277, and 1800497) in the human DRD2 gene are associated with decreased D2R expression and function. Immortalized renal proximal tubule cells (RPTCs) from subjects carrying D2R SNPs (RPTC-D2R SNPs) express less D2Rs than RPTCs carrying no D2R SNPs (RPTC-D2R WT) (62±4 vs 100±6%; P,0.04) and a pro-inflammatory and pro-fibrotic phenotype with markers of epithelial mesenchymal transition. RPTC-D2R SNPs showed increased apoptosis compared with RPTC-D2R WT (11± 0.8 vs 2.3±0.4% TUNEL positive cells, P,0.01, n=5/group). We hypothesized that the D2R regulates renal cell survival through effects on Wnt signaling. We found that Wnt3 expression was increased in RPTC-D2R SNPs compared with RPTC-D2R WT (mRNA: 2.6±0.35 vs 1±0.11 fold; P,0.05; protein: 133±4 vs ...
Tea catechins, a subclass of compounds in the flavonoid family, have been found to have several biologically beneficial properties including antioxidative effects [29, 30]. Flavonoids leading to cytoprotective effects against oxidative stress. Our findings, taken together with other observations [27, 30-32], indicate that catechin demonstrates renoprotective abilities in several models of renal disease, especially nephrolithic nephropathy. The experiments indicate that catechins can attenuate functional and immunohistochemical changes in the renal proximal tubular cell line NRK-52E treated with COM and kidneys of EG induced nephrolithic rats.. In our in vitro study, catechin attenuated the changes of mitochondrial membrane potential, and normalized expression of SOD, 4-HNE, cytochrome c, and cleaved caspase 3 in the renal proximal tubular cell line NRK-52E treated with COM.. The disappearance of TMRE in NRK-52E cells indicated mitochondrial collapse through depolarization of the mitochondrial ...
Although urinary sodium excretion is positively influenced by acute changes in renal perfusion pressure, micropuncture studies show highly conflicting results concerning the response of superficial proximal tubule sodium reabsorption to changes in renal perfusion pressure. In the present study, the changes of superficial proximal reabsorption to decreased and increased renal perfusion pressure were determined in rats by an in vivo microperfusion method. In vivo microperfusion was selected as the method to determine the proximal sodium reabsorption because this method made it possible to deliver a constant fluid and electrolyte load to the proximal tubule without the influence of possible changes of glomerular filtration rate. Renal perfusion pressure was decreased from normal pressure by inflating a suprarenal aortic cuff and was increased from the normal level by the occlusion of celiac and mesenteric arteries and the infrarenal aorta. Although fractional excretion of sodium (FENa) in the urine ...
Croft, Wayne D., Hills, Claire E., McCann, Eilish Clare, Bond, Michael, Esparza-Franco, Manuel A. , Bennett, Jeanette, Rand, D. A. (David A.), Davey, John and Ladds, Graham. (2013) A physiologically required G protein-coupled receptor (GPCR)-regulator of G protein signaling (RGS) interaction that compartmentalizes RGS activity. Journal of Biological Chemistry, Volume 288 (Number 38). pp. 27327-27342. ISSN 0021-9258 Bland, Rosemary, Zehnder, Daniel, Bennett, Jeanette, Markovic, Danijela and Hewison, Martin. (2012) Calcium sensing receptor-mediated regulation of vitamin D metabolism in a human proximal tubule cell line. Bone, Vol.51 (No.6). Article no.S29. ISSN 8756-3282 Hills, Claire E., Younis, Mustafa Y. G., Bennett, Jeanette, Siamantouras, Eleftherios, Liu, Kuo-Kang and Squires, Paul E.. (2012) Calcium-sensing receptor activation increases cell-cell adhesion and ß-cell function. Cellular Physiology and Biochemistry, Vol.30 (No.3). pp. 575-586. ISSN 1015-8987 Hills, Claire E., Bland, Rosemary, ...
Thyroid hormone has many actions on the kidney. Hypothyroidism can reduce glomerular filtration rate, renal blood flow, proximal tubule sodium absorption, impair renal acidification, BBMV Na-phosphate transport, NHE activity, and proximal and distal Na-K-ATPase activity (1, 3, 6, 25, 26, 28, 30). Conversely, hyperthyroidism increases proximal tubule sodium absorption, BBMV Na-phosphate transport and NHE activity, as well as proximal and distal tubule Na-K-ATPase activity (1, 6). While many of these alterations in proximal transport can be explained by the thyroid hormones effect on renal hemodynamics (16, 17, 21, 26), thyroid hormone has been shown to have a direct epithelial action to alter proximal tubular transport (13, 14, 24, 41).. Previous studies showed that the maturational increase in thyroid hormone likely is a significant factor affecting proximal tubule postnatal development. In neonatal rats, hypothyroidism prevents the maturational increase in NHE3 and reduces BBMV NHE activity ...
Na+ and volume homeostasis is controlled by the kidneys and key to blood pressure (BP) regulation. The kidneys respond to an increase in BP by decreasing Na+/H2O reabsorption (pressure natriuresis, diuresis) to decrease ECFV and BP. Conversely, a decrease in BP triggers Na+/H2O retaining mechanisms. Sodium transport can be regulated by altering transporter pool size, activity, and/or trafficking. In the proximal tubule (PT), trafficking is essential for the NHE3 and NaPi2 regulation. When BP increases, both retract away from the microvilli, NHE3 to the base, NaPi2 to endosomes. The aims of this dissertation were to determine: 1) the molecular basis of the differential trafficking patterns of NHE3 and NaPi2 during acute hypertension; 2) the role of acute AngII in the trafficking of PT sodium transporters; 3) the role of phosphorylation in the NHE3 and distal tubule NCC regulation during hypertension. Results: NHE3 and NaPi2 are segregated into domains: NHE3 to lipid rafts and NaPi2 to non-rafts. ...
Journal of Biomarkers is a peer-reviewed, Open Access journal that publishes original research articles, review articles, and clinical studies in all areas of biomarkers.
HRPTEpiC are isolated from human renal tissue, cryopreserved and characterized with antibodies against cytokeratin-18, -19 and vimentin. HRPTEpiC are guaranteed to further expand for 15 population doublings at the conditions specified in the Instruction for use sheet ...
Sigma-Aldrich offers abstracts and full-text articles by [Jianping Lu, Xinxiu Li, Mingcao Zhang, Zhaohong Chen, Yaping Wang, Caihong Zeng, Zhihong Liu, Huimei Chen].
Abstract Shenkang injection (SKI) is a classic prescription composed of Radix Astragali, rhubarb, Astragalus, Safflower, and Salvia. This treatment was approved by the State Food and Drug Administration of China in 1999 for treatment of chronic kidney diseases based on good efficacy and safety. This study aimed to investigate the protective effect of SKI against high glucose (HG)-induced renal tubular cell senescence and its underlying mechanism. Primary renal proximal tubule epithelial cells were cultured in (1) control medium (control group), medium containing 5 mmol/L glucose; (2) mannitol medium (mannitol group), medium containing 5 mmol/L glucose, and 25 mmol/L mannitol; (3) HG medium (HG group) containing 30 mmol/L glucose; (4) SKI treatment at high (200 mg/L), medium (100 mg/L), or low (50 mg/L) concentration in HG medium (HG+ SKI group); or (5) 200 mg/L SKI treatment in control medium (control+ SKI group) for 72 h. HG-induced senescent cells showed the emergence of senescence associated ...
1999 Schwerdt, G., R. Freudinger, S. Silbernagl and M. Gekle. Ochratoxin A-binding proteins in rat organs and plasma and in different cell lines of the kidney. Toxicology 135, 1-10, 1999 [abstract] Benesic, A., S. Eder, C. Sauvant, Schwerdt, G. and M. Gekle. Ochratoxin A induces proliferation of immortalized human kidney epithelial cells at low nanomolar concentrations by interference with cellular Ca2+-homeostasis. Proceedings 21st Mycotoxin-Workshop, Jena 130-138, 1999 Sauvant, C., S. Silbernagl and M. Gekle. Chronische Exposition von humanen Zellen des proximalen Tubulus mit nanomolaren Konzentrationen von freiem Ochratoxin A führt zu einer verminderten Expression des basolateralen Transportproteins für organische Anionen, OAT1 (Chronic exposure of human proximal tubule cells to nanomolar concentrations of free ochratoxin A leads to decreased expression of the basolateral anion transporter, OAT1) . Proceedings 21st Mycotoxin-Workshop, Jena 234-238, 1999 Schwerdt, G., R. Freudinger, S. ...
Objective: To investigate potential mechanisms of oxidative DNA damage in a rat model of type I diabetes and in murine proximal tubular epithelial cells (MCT) and primary culture of rat proximal tubular epithelial cells (RPTE).. Research Design and Methods: Phosphorylation of Akt and tuberin, 8-oxodG levels and OGG1 expression were measured in kidney cortical tissue of control and type I diabetic animals as well as in proximal tubular cells incubated with normal or high glucose.. Results: In the renal cortex of diabetic rats, the increase in Akt phosphorylation is associated with enhanced phosphorylation of tuberin, decreased OGG1 protein expression and 8-oxodG accumulation. Exposure of proximal tubular epithelial cells to high glucose (HG) causes a rapid increase in ROS generation that correlates with the increase in Akt and tuberin phosphorylation. HG also resulted in downregulation of OGG1 protein expression, paralleling its effect on Akt and tuberin. Inhibition of PI-3K/Akt significantly ...
BALB/c Mouse Proximal Tubular Epithelial Cells from Creative Bioarray are isolated from proximal tubular tissue of pathogen-free laboratory mice. BALB/c Mouse Proximal Tubular Epithelial Cells are grown in a T25 tissue culture flask pre-coated with gelatin-based coating solution for 2 min and incubated in Creative Bioarrays Culture Complete Growth Medium for 3-5 days. Cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 0.5x10^6 cells per ml and is delivered frozen. Cells can be expanded for 3-7 passages at a split ratio of 1:2 under the cell culture conditions specified by Creative Bioarray. Repeated freezing and thawing of cells is not recommended ...
Recent evidence suggests that mitochondria play roles in the pathogenesis of hypertension. However, it is not clear whether mitochondrial dysfunction precedes or follows the development of hypertension. We hypothesize that mitochondrial bioenergetics is altered in renal proximal tubule (RPT) cells from spontaneously hypertensive rats (SHRs) and this alteration may occur prior to the development of sustained hypertension. Using immortalized RPT cells from 4-8 week old normotensive Wistar-Kyoto (WKY) and SHRs, we measured oxygen consumption rate (OCR) and compared the parameters of cellular bioenergetic in these RPT cells. Basal OCR was significantly higher in RPT cells from SHRs than WKY rats (245.5±31.9 vs. 154.4±43.7 pmol/min per 20,000 cells, mean±SD, P,0.001);the ATP synthesis-coupled OCR (65.9±3.3% vs. 55.0±6.6%, of basal OCR, P,0.001), maximum respiration (609.0±120.4 vs. 215.7±65.5 pmol/min per 20,000 cells, P,0.001) and reserve respiration (147.8±39.0% vs, 40.0±15.4% of basal OCR ...
Exogenous cyclic 3,5-AMP (cAMP) and substances known to increase the intracellular concentration of this nucleotide (isoproterenol, theophylline, noradrenaline, lactate) were shown to inhibit the transport of fluorescein (a weak organic acid) into the rat renal proximal tubules at 20 degrees C. Carbacholine decreasing intracellular cAMP concentration stimulated the transport. Propranolol, a beta-adrenergic blockator, diminished significantly the inhibitory effect of noradrenaline on the transport. Lactate and carbacholine when added simultaneously, neutralize their action. The inhibitory action of intracellular cAMP on the transport is supposed to be a result of the diminition of a pool of endogenous weak organic acids which may take part in the exchange of diffusion with the marker anion across basal plasma membrane.
These results demonstrate that NHE8 protein is expressed on the brush-border membrane of rat kidney proximal tubules, extending from S1 to S3. Although NHE8 is expressed in all proximal tubules, a more intense NHE8 signal was evident in the deeper cortical and medullary proximal tubules compared with superficial proximal tubules. No other cell types in the kidney were detected to express NHE8 protein.. These findings correlate with results from a previous study, in which we used in situ hybridization to ascertain the localization of NHE8 in mouse kidney (6). In that study, NHE8 message was present in proximal tubules within the outer stripe of the outer medulla as well as a lower but significant expression diffusely throughout the cortex. Although the differential expression of NHE8 between the cortex and medulla appeared to be more pronounced than in the present study, the same tendency for higher expression in the deeper cortex/medulla was observed. Differences in technique (in situ ...
Sigma-Aldrich offers abstracts and full-text articles by [Mohammad A K Azad, Jesmin Akter, Kelly L Rogers, Roger L Nation, Tony Velkov, Jian Li].
Proximal cells produce and activate more TGF-β than distal cells. (A) Heat-activated conditioned medium from proximal and distal cells (4 × 104 cells/24-well
Kidney proximal tubule cells developed severe energy deficits during hypoxia/reoxygenation not attributable to cellular disruption lack of purine precursors the mitochondrial permeability transition or loss of cytochrome from your intermembrane space into the cytosol (5). hypoxia and ischemia (1 3 However the proximate events that lead to the MPT and loss of cytochrome are unclear and are subjects of ongoing investigation. We have reported that cells in freshly isolated kidney proximal tubules exhibit profound functional deficits of their mitochondria when they MK-0822 are reoxygenated after hypoxic incubation (6). The defect is usually characterized by failure of oxidative phosphorylation in cells that are MK-0822 normally intact as indicated by structural biochemical and functional criteria and is partially ameliorated by prior MK-0822 treatment with chemical inhibitors of the MPT (6) but not by antioxidants or redox state modification (J.M.W. unpublished data). We have now more completely ...
The regulation of the actions of albumin in proximal tubular epithelial cells by endocytosis and the role played by adaptor protein disabled 2 ...
A key requirement for such a device is the formation of a "living membrane" that consists of a tight kidney cell layer on artificial membrane surfaces and can transport molecules from one side to the other. In their presentation, Dimitrios Stamatialis, PhD from University of Twente in The Netherlands, Roos Masereeuw, PhD from University of Utrecht in The Netherlands discussed achieving this using conditionally immortalized human renal proximal tubular epithelial cells (ciPTECs) on polyethersulfone-based hollow fiber membranes ...
TY - JOUR. T1 - A renal dopamine-1 receptor defect in two genetic models of hypertension. AU - Felder, R. A.. AU - Kinoshita, S.. AU - Sidhu, A.. AU - Ohbu, K.. AU - Kaskel, Frederick J.. PY - 1990. Y1 - 1990. N2 - Dopamine (DA), via DA-1 receptors, regulates Na+ transport in the kidneys. Dopamine is synthesized from L-DOPA in the proximal tubule and presumably secreted as an autocrine/paracrine substance to stimulate DA-1 receptors localized on proximal tubular cells. We have previously reported the presence of DA-1 receptors in renal cortical homogenates and on the isolated proximal tubule of the rat and rabbit, consistent with the dopamine autocrine/paracrine model. We have localized DA-1 receptors in the proximal straight tubule of the rabbit, and in the cortical collecting duct of the rabbit and rat, but not in the distal collecting tubule or the cortical thick ascending loop of Henle. The presence of functional DA-1 receptors has been substantiated by the coexistence of DA-1 ...
Background: Prostate cancer (PC) is a radiosensitive disease. In addition to external beam and brachytherapy, systemic bone-seeking radioisotopes have been utilized clinically for many years, with radium-223 leading to overall survival benefits. However, the available beta and alpha emitting particles in the clinic target tumors / stroma in bone only. Biologically targeted radiopharmaceutical treatment has shown promising results in our earlier multiple clinical trials using beta-emitting anti-PSMA therapy with Leutetium-177 radiolabeled anti-PSMA monoclonal antibody (mAb) J591, with accurate targeting and tumor response, but treatment is limited by myelosuppression. Both beta and alpha-small molecule PSMA ligand directed therapy has been utilized anecdotally outside of the US, but has not been systemically studied. Based upon their biodistribution, targeting of salivary glands and potentially proximal renal tubules may pose longer term toxicity issues. mAb-based targeted alpha-particle delivery ...
Kim-1/Tim-1 is an apoptotic-cell phagocytosis and scavenger receptor that is most highly upregulated in proximal tubular epithelium in acute and chronic kidney injury. While Kim-1/Tim-1 has been proposed to be a costimulatory ...
Barati, M., Ketchem, C., Merchant, M., Kusiak, W., Jose, P., Weinman, E., LeBlanc, A., Lederer, E., & Khundmiri, S. (2017). Loss of NHERF-1 Expression Prevents Dopamine-Mediated Na-K-ATPase Regulation in Renal Proximal Tubule Cells from Rat Models of Hypertension: Aged F344 Rats and Spontaneously Hypertensive Rats.. American Journal of Physiology. Cell Physiology, 313 (2). http://dx.doi.org/10.1152/ajpcell.00219.2016 ...
In this study, we provide the first evidence that Na,K-ATPase is a target of TGF-β1 signaling during the induction of EMT. We show that following the treatment of renal proximal tubule cells with TGF-β1, NaK-β1 surface expression is reduced before well-characterized EMT markers, such as E-cadherin, and the induction of fibronectin and α-SMA. We validated the significance of NaK-β1 in the induction of EMT by two complementary approaches. RNAi-mediated specific knockdown of NaK-β1 resulted in the loss of the epithelial phenotype, whereas the ectopic expression of NaK-β1 delayed the induction of a fibroblastic phenotype and reduced the levels of fibronectin and α-SMA following TGF-β1 treatment. We further show that NaK-β1 reduction is a common event in EMT irrespective of the type of EMT.. RNAi-mediated specific knockdown of NaK-β1 is sufficient to induce a fibroblastic phenotype of LLC-PK1 cells. We have shown earlier that NaK-β1 expression is reduced in a wide variety of poorly ...
One of the major tasks of the renal proximal tubule (PT) is to secrete acid into the tubule lumen, thereby reabsorbing approximately 80% of the filtered bicarbonate (HCO3(-)), as well as generating "new HCO3(-)" for regulating blood pH. In the tubular lumen, filtered HCO3(-) combines with H(+) in a reaction catalyzed by CA IV. The CO2 thus produced rapidly diffuses into the tubular cells and is combined with water to produce intracellular H(+) and HCO3(-), catalyzed by soluble cytoplasmic CA II. HCO3(-) is then cotransported with Na(+) into blood via the NBC-1. The intracellular H(+) produced by CA II is secreted into the tubular lumen predominantly via the NHE-3. The PT creates the "new HCO3(-)" by taking glutamine and metabolizing it to two molecules each of NH4(+) and HCO3(-). The NH4(+) is excreted into the tubular lumen, and the HCO3(-) , which is "new HCO3(-) ," is returned to the blood, where it replaces the HCO3(-) lost earlier in the titration of nonvolatile acids ...
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In the present experiments on microdissected tubules of rabbit kidney we present a refined stop-flow method for determining the rate of HCO3- absorption (J(HCO3)) or H+ secretion (JH) that can be applied to isolated microperfused tubules. Using the pH-sensitive indicator dye BCECF (2,7-bis [2-carboxyethyl]-5[6]-carboxyfluorescein) the luminal perfusate pH is continuously ...
TY - JOUR. T1 - Action of EGF and PGE2 on basolateral organic anion uptake in rabbit proximal renal tubules and hOAT1 expressed in human kidney epithelial cells. AU - Sauvant, C.. AU - Hesse, D.. AU - Holzinger, H.. AU - Evans, K. K.. AU - Dantzler, William H. AU - Gekle, M.. PY - 2004/4. Y1 - 2004/4. N2 - We recently showed that, in a proximal tubule cell line (opossum kidney cells), epithelial growth factor (EGF) stimulates basolateral organic anion transport (OAT) via ERK1/2, arachidonic acid, phospholipase A2, and generation of prostaglandins. PGE2 binds the prostanoid receptor and, thus, activates adenylate cyclase and PKA, which stimulate basolateral organic anion uptake. In the present study, we investigated whether this regulatory cascade is also true 1) for ex vivo conditions in isolated renal proximal (S2) tubules from rabbit and 2) in a human renal epithelial cell line stably expressing human OAT1 (IHKE-hOAT1). EGF activated ERK1/2 in S2 tubules and IHKE-hOAT1, and, in both cases, ...
The effects of angiotensin II on total ammonia (tNH3) production and net secretion were investigated using in vitro microperfused mouse S2 proximal tubule segments incubated in Krebs-Ringer bicarbonate buffer containing 0.5 mM L-glutamine. Basolateral exposure of mouse S2 segments to 10(-11), 10(-10 …
Correspondence to Dr. Frans G. M. Russel, Department of Pharmacology and Toxicology 233, Nijmegen Center for Molecular Life Sciences, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. Phone: +31-24-3613691/3616892; Fax: +31-24-3614214; E-mail: F.Russel{at}farm.kun.nl ...
TY - JOUR. T1 - Activation of BNIP3-mediated mitophagy protects against renal ischemia-reperfusion injury. AU - Tang, Chengyuan. AU - Han, Hailong. AU - Liu, Zhiwen. AU - Liu, Yuxue. AU - Yin, Lijun. AU - Cai, Juan. AU - He, Liyu. AU - Liu, Yu. AU - Chen, Guochun. AU - Zhang, Zhuohua. AU - Yin, Xiao-Ming. AU - Dong, Zheng. PY - 2019/9/12. Y1 - 2019/9/12. N2 - Acute kidney injury (AKI) is a syndrome of abrupt loss of renal functions. The underlying pathological mechanisms of AKI remain largely unknown. BCL2-interacting protein 3 (BNIP3) has dual functions of regulating cell death and mitophagy, but its pathophysiological role in AKI remains unclear. Here, we demonstrated an increase of BNIP3 expression in cultured renal proximal tubular epithelial cells following oxygen-glucose deprivation-reperfusion (OGD-R) and in renal tubules after renal ischemia-reperfusion (IR)-induced injury in mice. Functionally, silencing Bnip3 by specific short hairpin RNAs in cultured renal tubular cells reduced ...
The transport characteristics of amino acids in primary cell cultures from the proximal tubule of human adults (AHKE cells) were examined, using alpha-aminoisobutyric acid (AIB) and beta-alanine as representatives of alpha- and beta-amino acids, respectively. The Na(+)-gradient dependent influx of AIB occurred by a single, saturable transport system, whereas the Na(+)-gradient dependent uptake data for beta-alanine could be described in terms of two-independent transport components as well as one-transport one-leak model with identical kinetic constants for the high-affinity system. Competition experiments revealed that all the neutral amino acids tested reduced the uptake of AIB, whereas there was no effect of taurine, L-aspartic acid, and L-arginine. By contrast, the influx of beta-alanine was only drastically reduced by beta-amino acids, whereas the inhibition by neutral alpha-amino acids was relatively low. Nor did L-arginine and L-aspartic acid affect the uptake of beta-alanine into AHKE ...
DI-fusion, le Dépôt institutionnel numérique de lULB, est loutil de référencementde la production scientifique de lULB.Linterface de recherche DI-fusion permet de consulter les publications des chercheurs de lULB et les thèses qui y ont été défendues.
A: In adult Hsd11b2tm1Yko/Hsd11b2tm1Yko mice, the distal tubules (d) show considerable enlargement: the tubular diameter is increased threefold compared with the distal tubule in the wild-type mice (B). Note that the proximal tubules (p) and glomeruli (g) are unaffected in the mutant mice. C: Hyperplasia is apparent in the mutant mouse distal tubule (d), which shows over 20 cells in the cross-sectional profile compared with (D) the four to six cells observed in the wild-type. E: The distal tubular enlargement (d) is evident in mutant mice at the anatomical origin of the distal tubule at the juxtaglomerular apparatus (j) adjacent to the glomerulus (g), compared with (F) the nonenlarged wild-type distal tubule. md, macula densa. G: Proximal tubule (p) is normal in mutant kidneys. Mutant proximal tubules are indistinguishable from (H) wild-type proximal tubules. The tubules are of similar diameter and cell number in cross section. They are lined by cubiodal cells with granular cytoplasm and a brush ...
Renal proximal tubules are the major target of toxicant and ischemic injury in the kidney. Because mitochondria play a crucial role in ATP generation and survival of RPTCs, mitochondrial dysfunction caused by toxicant exposure or ischemia leads to RPTC injury and death and plays a major role in the development of acute kidney injury (Abid et al., 2005; Cachofeiro et al., 2008; Koyner et al., 2008). Oxidative stress is believed to be an important mechanism of toxicant- and ischemia-induced injury to RPTCs (Abid et al., 2005; Cachofeiro et al., 2008; Koyner et al., 2008). Therefore, we hypothesized that GT3, a potent antioxidant, may limit or prevent mitochondrial dysfunction, energy deficits, and cell death caused by oxidant exposure. Our data demonstrate that GT3 is taken up by RPTCs by a process that is saturated within 24 h, which suggests that GT3 is not taken up by simple diffusion. The saturation of GT3 uptake is not caused by depletion of GT3 from the cell culture media. At this point of ...
Endothelial-mesenchymal transition (EndoMT) has been shown to be a major source of myofibroblasts, contributing to kidney fibrosis. However, in vitro study of endothelial cells often relies on culture of isolated primary endothelial cells due to the unavailability of endothelial cell lines. Our recent study suggested that peritubular endothelial cells could contribute to kidney fibrosis through EndoMT. Therefore, successful isolation and culture of mouse peritubular endothelial cells could provide a new platform for studying kidney fibrosis. This study describes an immunomagnetic separation method for the isolation of mouse renal peritubular endothelial cells using anti-CD146 MicroBeads, followed by co-culture with mouse renal proximal tubular epithelial cells to maintain endothelial phenotype. Flow cytometry showed that after isolation and two days of culture, about 95% of cells were positive for endothelial-specific marker CD146. The percentage of other cells, including dendritic cells (CD11c) and
Chatterjee, S.; Ghosh, N.; Castiglione, E.; Kwiterovich, P.O., 1988: Regulation of glycosphingolipid glycosyltransferase by low density lipoprotein receptors in cultured human proximal tubular cells
Multidrug resistance protein 4 (MRP4; ABCC4) is an ATP binding cassette transporter that facilitates the excretion of bile salt conjugates and other conjugated steroids in hepatocytes and renal proximal tubule epithelium. MRP4/Mrp4 undergoes adaptive upregulation in response to oxidative and cholest …
0258] U.S. Pat. No. 5,429,938 [0259] Ryan M J, Johnson O, Kirk I, Fuerstenberg S M, Zager R A, Torok-Storb B. HK-2: an immortalized proximal tubule epithelial cell line from normal adult human kidney. Kidney Int 1994; 45(1):48-57. [0260] Beacham D A, Amatangelo M D, Cukierman E. Preparation of extracellular matrices produced by cultured and primary fibroblasts. Curr Protoc Cell Biol 2006; Supplement 33(Chapter 10:Unit 10.9): 10.9.1-10.9.21. [0261] Sadoni N, Langer S, Fauth C, Bennardi O, Cremer T, Turner B M, et al. Nuclear organization of mammalian genomes: polar chromosome territories build up functionally distinct higher order compartments. J Cell Biol 1999; 146(6): 1211-1226. [0262] Karihaloo A, Nickel C, Cantley L G. Signals which build a tubule. Nephron Exp Nephrol 2005; 100(1):e40-45. [0263] Schumacher K M, Phua S C, Schumacher A, Ying J Y. Controlled formation of biological tubule systems in extracellular matrix gels in vitro. Kidney Int 2008; 73(10): 1187-1192. [0264] Han H J, Sigurdson ...
Looking for Kidney tubules? Find out information about Kidney tubules. One of the glandular tubules which elaborate urine in the kidneys Explanation of Kidney tubules
Cells and reagents. Human proximal tubular epithelial cells. HK-2 cells, which have been well characterized (60), were obtained from the American Type Culture Collection (ATCC). Monolayers of HK-2 cells were grown on 6-well plates (Corning-Costar, Corning Life Sciences) that were precoated with 5 μg/cm2 type 1 collagen (Rat Tail Collagen Type 1, Invitrogen, Thermo Fisher Scientific) and incubated at 37°C with 5% CO2 and 95% air in keratinocyte serum-free medium (K-SFM) (Gibco, Invitrogen, Thermo Fisher Scientific) supplemented with recombinant human epidermal growth factor (5 ng/ml) and bovine pituitary extract (50 μg/ml). Medium was exchanged at 48-hour intervals, and the cells were not use beyond 25 to 30 passages.. Human immunoglobulin FLCs. Six unique human monoclonal FLCs (three and three) were purified using standard methods from the urine of patients with multiple myeloma, light chain proteinuria, and clinical evidence of significant renal damage that was presumed to be cast ...
Cells and reagents. Human proximal tubular epithelial cells. HK-2 cells, which have been well characterized (60), were obtained from the American Type Culture Collection (ATCC). Monolayers of HK-2 cells were grown on 6-well plates (Corning-Costar, Corning Life Sciences) that were precoated with 5 μg/cm2 type 1 collagen (Rat Tail Collagen Type 1, Invitrogen, Thermo Fisher Scientific) and incubated at 37°C with 5% CO2 and 95% air in keratinocyte serum-free medium (K-SFM) (Gibco, Invitrogen, Thermo Fisher Scientific) supplemented with recombinant human epidermal growth factor (5 ng/ml) and bovine pituitary extract (50 μg/ml). Medium was exchanged at 48-hour intervals, and the cells were not use beyond 25 to 30 passages.. Human immunoglobulin FLCs. Six unique human monoclonal FLCs (three and three) were purified using standard methods from the urine of patients with multiple myeloma, light chain proteinuria, and clinical evidence of significant renal damage that was presumed to be cast ...
TASK-2 (KCNK5) two-pore domain, pH-sensitive, voltage-insensitive, outward rectifying K+ channel (K+ , Rb+ ,, Cs+ , NH4+ , Na+ ≈ Li+), present in renal epithelia. Regulated [inhibited] via group 1 metabolotropic glutamate receptors and by inositol phosphates (Chemin et al., 2003). TASK-2 gating is controlled by changes in both extra- and intracellular pH through separate sensors: arginine 224 and lysine 245, located at the extra- and intracellular ends of transmembrane domain 4, respectively. TASK-2 is inhibited by a direct effect of CO2 and is regulated by and interacts with G protein subunits. TASK-2 takes part in regulatory adjustments and is a mediator in the chemoreception process in neurons of the retrotrapezoid nucleus where its pHi sensitivity could be important in regulating excitability and therefore signalling of the O2/CO2 status. Extracellular pH increases, brought about by HCO3- efflux from proximal tubule epithelial cells may couple to TASK-2 activation to maintain ...
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