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Culture proximal tubular cell

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https://www.readbyqxmd.com/read/27880866/tgf-%C3%AE-1-stimulates-movement-of-renal-proximal-tubular-epithelial-cells-in-a-three-dimensional-cell-culture-via-an-autocrine-tgf-%C3%AE-2-production
#1
Deyi Luo, Qiunong Guan, Kunjie Wang, Christopher Y C Nguan, Caigan Du
TGF-βs are multifunctional cytokines, but their roles in human renal homeostasis are not fully understood. This study investigated the role of TGF-β1 in the movement of human renal proximal tubular epithelial cells (PTECs) in a three-dimensional (3D) model. HKC-8 cells, a human PTEC line, were grown in a 3D collagen culture system. Cell movement was observed under a microscope. The gene expression was examined using PCR Arrays or qRT-PCR, and protein levels by Western blot. Here, we showed that the tight junction structure formed between adjacent cells of a HKC-8 cell colony in 3D cultures, and TGF-β1 stimulated their movement, evidenced by the appearance of fingerlike pseudopodia in the leader cells at the edge of the colonies...
November 20, 2016: Experimental Cell Research
https://www.readbyqxmd.com/read/27872161/identification-of-urinary-exosomal-non-coding-rnas-as-novel-biomarkers-in-chronic-kidney-disease
#2
Rimpi Khurana, Glory Ranches, Simon Schafferer, Melanie Lukasser, Michael Rudnicki, Gert Mayer, Alexander Hüttenhofer
In chronic kidney disease (CKD), the decline in glomerular filtration rate is associated with increased morbidity and mortality and thus poses a major challenge for healthcare systems. While the contribution of tissue-derived miRNAs and mRNAs to CKD progression has extensively been studied, little is known about the role of urinary exosomes and their association with CKD. Exosomes are small, membrane-derived endocytic vesicles that contribute to cell-to-cell communication and are present in various body fluids, such as blood or urine...
November 21, 2016: RNA
https://www.readbyqxmd.com/read/27867451/thioredoxin-interacting-protein-mediates-nlrp3-inflammasome-activation-involved-in-the-susceptibility-to-ischemic-acute-kidney-injury-in-diabetes
#3
Ye Da Xiao, Ya Yi Huang, Hua Xin Wang, Yang Wu, Yan Leng, Min Liu, Qian Sun, Zhong-Yuan Xia
Kidney in diabetic state is more sensitive to ischemic acute kidney injury (AKI). However, the underlying mechanisms remain unclear. Herein, we examined the impact of diabetes mellitus on thioredoxin-interacting protein (TXNIP) expression and whether mediated NLRP3 activation was associated with renal ischemia/reperfusion- (I/R-) induced AKI. In an in vivo model, streptozotocin-induced diabetic rats showed higher susceptibility to I/R injury with increased TXNIP expression, which was significantly attenuated by resveratrol (RES) treatment (10 mg/kg intraperitoneal daily injection for 7 consecutive days prior to I/R induction)...
2016: Oxidative Medicine and Cellular Longevity
https://www.readbyqxmd.com/read/27861627/caspase-dependent-and-caspase-independent-pathways-are-involved-in-cadmium-induced-apoptosis-in-primary-rat-proximal-tubular-cell-culture
#4
Gang Liu, Hui Zou, Tongwang Luo, Mengfei Long, Jianchun Bian, Xuezhong Liu, Jianhong Gu, Yan Yuan, Ruilong Song, Yi Wang, Jiaqiao Zhu, Zongping Liu
We designed this study to investigate whether cadmium induces caspase-independent apoptosis and to investigate the relationship between the caspase-dependent and caspase-independent apoptotic pathways. Cadmium (1.25-2.5 μM) induced oxidative stress in rat proximal tubular (rPT) cells, as seen in the reactive oxygen species levels; N-acetylcysteine prevented this. Cyclosporin A (CsA) prevented mitochondrial permeability transition pore opening and apoptosis; there was mitochondrial ultrastructural disruption, mitochondrial cytochrome c (cyt c) translocation to the cytoplasm, and subsequent caspase-9 and caspase-3 activation...
2016: PloS One
https://www.readbyqxmd.com/read/27834856/stearoyl-coa-desaturase-1-protects-cells-against-lipotoxicity-mediated-apoptosis-in-proximal-tubular-cells
#5
Tamaki Iwai, Shinji Kume, Masami Chin-Kanasaki, Shogo Kuwagata, Hisazumi Araki, Naoko Takeda, Takeshi Sugaya, Takashi Uzu, Hiroshi Maegawa, Shin-Ichi Araki
Saturated fatty acid (SFA)-related lipotoxicity is a pathogenesis of diabetes-related renal proximal tubular epithelial cell (PTEC) damage, closely associated with a progressive decline in renal function. This study was designed to identify a free fatty acid (FFA) metabolism-related enzyme that can protect PTECs from SFA-related lipotoxicity. Among several enzymes involved in FFA metabolism, we identified stearoyl-CoA desaturase-1 (SCD1), whose expression level significantly decreased in the kidneys of high-fat diet (HFD)-induced diabetic mice, compared with non-diabetic mice...
November 9, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27821630/mitochondrial-nadp-dependent-isocitrate-dehydrogenase-deficiency-exacerbates-mitochondrial-and-cell-damage-after-kidney-ischemia-reperfusion-injury
#6
Sang Jun Han, Hee-Seong Jang, Mi Ra Noh, Jinu Kim, Min Jung Kong, Jee In Kim, Jeen-Woo Park, Kwon Moo Park
Mitochondrial NADP(+)-dependent isocitrate dehydrogenase (IDH2) catalyzes the oxidative decarboxylation of isocitrate to α-ketoglutarate, synthesizing NADPH, which is essential for mitochondrial redox balance. Ischemia-reperfusion (I/R) is one of most common causes of AKI. I/R disrupts the mitochondrial redox balance, resulting in oxidative damage to mitochondria and cells. Here, we investigated the role of IDH2 in I/R-induced AKI. I/R injury in mice led to the inactivation of IDH2 in kidney tubule cells. Idh2 gene deletion exacerbated the I/R-induced increase in plasma creatinine and BUN levels and the histologic evidence of tubule injury, and augmented the reduction of NADPH levels and the increase in oxidative stress observed in the kidney after I/R...
November 7, 2016: Journal of the American Society of Nephrology: JASN
https://www.readbyqxmd.com/read/27815996/human-airway-organoid-engineering-as-a-step-toward-lung-regeneration-and-disease-modeling
#7
Qi Tan, Kyoung Moo Choi, Delphine Sicard, Daniel J Tschumperlin
Organoids represent both a potentially powerful tool for the study cell-cell interactions within tissue-like environments, and a platform for tissue regenerative approaches. The development of lung tissue-like organoids from human adult-derived cells has not previously been reported. Here we combined human adult primary bronchial epithelial cells, lung fibroblasts, and lung microvascular endothelial cells in supportive 3D culture conditions to generate airway organoids. We demonstrate that randomly-seeded mixed cell populations undergo rapid condensation and self-organization into discrete epithelial and endothelial structures that are mechanically robust and stable during long term culture...
October 28, 2016: Biomaterials
https://www.readbyqxmd.com/read/27799484/renal-fanconi-syndrome-and-hypophosphatemic-rickets-in-the-absence-of-xenotropic-and-polytropic-retroviral-receptor-in-the-nephron
#8
Camille Ansermet, Matthias B Moor, Gabriel Centeno, Muriel Auberson, Dorothy Zhang Hu, Roland Baron, Svetlana Nikolaeva, Barbara Haenzi, Natalya Katanaeva, Ivan Gautschi, Vladimir Katanaev, Samuel Rotman, Robert Koesters, Laurent Schild, Sylvain Pradervand, Olivier Bonny, Dmitri Firsov
Tight control of extracellular and intracellular inorganic phosphate (Pi) levels is critical to most biochemical and physiologic processes. Urinary Pi is freely filtered at the kidney glomerulus and is reabsorbed in the renal tubule by the action of the apical sodium-dependent phosphate transporters, NaPi-IIa/NaPi-IIc/Pit2. However, the molecular identity of the protein(s) participating in the basolateral Pi efflux remains unknown. Evidence has suggested that xenotropic and polytropic retroviral receptor 1 (XPR1) might be involved in this process...
October 31, 2016: Journal of the American Society of Nephrology: JASN
https://www.readbyqxmd.com/read/27780843/binding-of-anti-dsdna-antibodies-to-proximal-tubular-epithelial-cells-contribute-s-to-renal-tubulo-interstitial-inflammation
#9
Susan Yung, Claudia Yc Ng, Kin Yi Au, Kwok Fan Cheung, Qing Zhang, Chenzhu Zhang, Desmond Yh Yap, Mel Km Chau, Tak Mao Chan
Immune deposits are often observed along the tubular basement membrane in patients with lupus nephritis, but the role of anti-dsDNA antibody deposition on tubulo-interstitial inflammation remains to be investigated. We examined the effect of human polyclonal anti-dsDNA antibodies on inflammatory processes in cultured proximal renal tubular epithelial cells (HK-2 cells) and their association with serum levels of IL-6, IL-8 and MCP-1 in patients. Binding of anti-dsDNA antibodies to HK-2 cells was investigated by cellular ELISA, flow cytometry and immunohistochemistry...
October 25, 2016: Clinical Science (1979-)
https://www.readbyqxmd.com/read/27776163/potential-use-of-autologous-renal-cells-from-diseased-kidneys-for-the-treatment-of-renal-failure
#10
Sunil K George, Mehran Abolbashari, John D Jackson, Tamer Aboushwareb, Anthony Atala, James J Yoo
Chronic kidney disease (CKD) occurs when certain conditions cause the kidneys to gradually lose function. For patients with CKD, renal transplantation is the only treatment option that restores kidney function. In this study, we evaluated primary renal cells obtained from diseased kidneys to determine whether their normal phenotypic and functional characteristics are retained, and could be used for cell therapy. Primary renal cells isolated from both normal kidneys (NK) and diseased kidneys (CKD) showed similar phenotypic characteristics and growth kinetics...
2016: PloS One
https://www.readbyqxmd.com/read/27760765/deletion-of-protein-kinase-c-%C3%AE%C2%B5-attenuates-mitochondrial-dysfunction-and-ameliorates-ischemic-renal-injury
#11
Grazyna Nowak, Diana Takacsova Bakajsova, Judit Megyesi
Previously, we documented that activation of protein kinase C-ε (PKC-ε) mediates mitochondrial dysfunction in cultured renal proximal tubule cells (RPTC). This study tested whether deletion of the PKC-ε decreases dysfunction of renal cortical mitochondria and improves kidney function after renal ischemia. PKC-ε levels in mitochondria of ischemic kidneys increased 24h after ischemia. Complex I- and complex II-coupled state 3 respirations were reduced 44% and 27%, respectively, in wild-type (WT) but unchanged and increased in PKC-ε-deficient (KO) mice after ischemia...
October 19, 2016: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/27633115/mechanism-of-tacrolimus-induced-chronic-renal-fibrosis-following-transplantation-is-regulated-by-ox-ldl-and-its-receptor-lox-1
#12
Shi Deng, Tao Jin, Li Zhang, Hong Bu, Peng Zhang
Chronic renal allograft dysfunction (CRAD) is the most common cause of graft failure following renal transplantation. However, the underlying mechanisms remain to be fully elucidated. Immunosuppressants and hyperlipidemia are associated with renal fibrosis following long‑term use. The present study aimed to determine the effects of tacrolimus (FK506) and lipid metabolism disorder on CRAD. In vitro and in vivo models were used for this investigation. Cells of the mouse proximal renal tubular epithelial cell strain, NRK‑52E, were cultured either with oxidized low‑density lipoprotein (ox‑LDL), FK506, ox‑LDL combined with FK506, or vehicle, respectively...
November 2016: Molecular Medicine Reports
https://www.readbyqxmd.com/read/27622990/induction-of-microrna-17-5p-by-p53-protects-against-renal-ischemia-reperfusion-injury-by-targeting-death-receptor-6
#13
Jielu Hao, Qingqing Wei, Shuqin Mei, Lin Li, Yunchao Su, Changlin Mei, Zheng Dong
Renal ischemia-reperfusion injury is a leading cause of acute kidney injury; the pathogenesis of which remains poorly understood and effective therapies are still lacking. Here we tested whether microRNAs, identified as critical regulators of cell health and disease, are involved in this process. We found that miR-17-5p was significantly up-regulated during renal ischemia-reperfusion injury in mice and during hypoxia in cultured renal tubular cells. In cultured cells, miR-17-5p directly inhibited the expression of death receptor 6 (DR6) and attenuated apoptosis during hypoxia...
September 9, 2016: Kidney International
https://www.readbyqxmd.com/read/27599751/mxra5-is-a-tgf-%C3%AE-1-regulated-human-protein-with-anti-inflammatory-and-anti-fibrotic-properties
#14
Jonay Poveda, Ana B Sanz, Beatriz Fernandez-Fernandez, Susana Carrasco, Marta Ruiz-Ortega, Pablo Cannata-Ortiz, Alberto Ortiz, Maria D Sanchez-Niño
Current therapy for chronic kidney disease (CKD) is unsatisfactory because of an insufficient understanding of its pathogenesis. Matrix remodelling-associated protein 5 (MXRA5, adlican) is a human protein of unknown function with high kidney tissue expression, not present in rodents. Given the increased expression of MXRA5 in injured tissues, including the kidneys, we have suggested that MXRA5 may modulate kidney injury. MXRA5 immunoreactivity was observed in tubular cells in human renal biopsies and in urine from CKD patients...
September 6, 2016: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/27591740/a-kidney-injury-molecule-1-kim-1-gene-reporter-in-a-mouse-artificial-chromosome-the-responsiveness-to-cisplatin-toxicity-in-immortalized-mouse-kidney-s3-cells
#15
Kenji Kokura, Yasushi Kuromi, Takeshi Endo, Naohiko Anzai, Yasuhiro Kazuki, Mitsuo Oshimura, Tetsuya Ohbayashi
BACKGROUND: Kidney injury molecule-1 (Kim-1) has been validated as a urinary biomarker for acute and chronic renal damage. The expression of Kim-1 mRNA is also activated by acute kidney injury induced by cisplatin in rodents and humans. To date, the measurement of Kim-1 expression has not fully allowed the detection of in vitro cisplatin nephrotoxicity in immortalized culture cells such as human kidney-2 cells and immortalized proximal tubular epithelial cells. METHODS: We measured the augmentation of Kim-1 mRNA expression after cisplatin addition using immortalized S3 cells established from the kidneys of transgenic mice harboring temperature-sensitive large T antigen from Simian virus 40...
September 3, 2016: Journal of Gene Medicine
https://www.readbyqxmd.com/read/27591086/microrna148b-3p-inhibits-mtorc1-dependent-apoptosis-in-diabetes-by-repressing-tnfr2-in-proximal-tubular-cells
#16
Shogo Kuwagata, Shinji Kume, Masami Chin-Kanasaki, Hisazumi Araki, Shinichi Araki, Jun Nakazawa, Takeshi Sugaya, Daisuke Koya, Masakazu Haneda, Hiroshi Maegawa, Takashi Uzu
Hypoxia causes proximal tubular cell damage in diabetes, even though proximal tubular cells have an adaptive system to combat hypoxia involving induction of hypoxia factor-1 (HIF-1) and inhibition of mechanistic target of rapamycin complex 1 (mTORC1). Here, we examined the interference effect of altered glucose and lipid metabolism on the hypoxia responses in proximal tubular cells. In culture, hypoxia alone induced HIF-1 and inhibited mTORC1, preventing death in proximal tubular cells. However, hypoxia with high glucose and palmitate increased mTORC1 activity and promoted apoptosis in proximal tubular cells, which was inhibited by pharmacological and genetic inactivation of mTORC1...
August 30, 2016: Kidney International
https://www.readbyqxmd.com/read/27521227/effects-of-shiga-toxin-2-on-cellular-regeneration-mechanisms-in-primary-and-three-dimensional-cultures-of-human-renal-tubular-epithelial-cells
#17
Laura B Márquez, Alicia Araoz, Horacio A Repetto, Fernando R Ibarra, Claudia Silberstein
Shiga toxin (Stx)-producing Escherichia coli (STEC) causes post-diarrheal Hemolytic Uremic Syndrome (HUS), which is one of the most common causes of acute renal failure in children in Argentine. The aim of the present work was to study the effects of Shiga toxin type 2 (Stx2) on regenerative mechanisms of primary cultures of human cortical renal tubular epithelial cells (HRTEC) and three-dimensional (3D) cultures of HRTEC. Primary cultures of HRTEC were able to develop tubular structures when grown in matrigel, which showed epithelial cells surrounding a central lumen resembling the original renal tubules...
October 2016: Microbial Pathogenesis
https://www.readbyqxmd.com/read/27470548/sirt1-activator-ameliorates-the-renal-tubular-injury-induced-by-hyperglycemia-in-vivo-and-in-vitro-via-inhibiting-apoptosis
#18
Xue-Ling Wang, Li-Yan Wu, Long Zhao, Li-Na Sun, Hai-Ying Liu, Gang Liu, Guang-Ju Guan
We aimed to explore the role of SIRT1 in apoptosis in human kidney proximal tubule epithelial (HK-2) cells, and to determine whether resveratrol (RSV, a SIRT1 activator) could ameliorate apoptosis in rats with streptozotocin-induced diabetes mellitus (DM) and/or in high glucose (HG, 30mM) - stimulated HK-2 cells. Rats were distributed randomly into three groups: 1) control group, 2) DM group, and 3) DM with RSV group (DM+RSV; rats treated with 30mg/kg/d of RSV for 16 weeks). The physical, biochemical, and morphological parameters were then examined...
June 21, 2016: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/27451286/stress-response-gene-nupr1-alleviates-cyclosporin-a-nephrotoxicity-in-vivo
#19
Pierre Galichon, Aurélien Bataille, Sophie Vandermeersch, Morgane Wetzstein, Yi-Chun Xu-Dubois, David Legouis, Alexandre Hertig, David Buob, Sandrine Placier, Naïke Bigé, Guillaume Lefevre, Chantal Jouanneau, Caroline Martin, Juan Lucio Iovanna, Eric Rondeau
Acute tubular damage is a major cause of renal failure, especially at the early phase of kidney transplant when ischemia-reperfusion injury and cyclosporin A toxicity may coexist. The mechanisms of the latter are largely unknown. Using an mRNA microarray on microdissected tubules from a rat model of cyclosporin A toxicity to describe the related epithelial-specific transcriptional signature in vivo, we found that cyclosporin A induces pathways dependent on the transcription factor ATF4 and identified nuclear protein transcriptional regulator 1 (Nupr1), a stress response gene induced by ATF4, as the gene most strongly upregulated...
July 22, 2016: Journal of the American Society of Nephrology: JASN
https://www.readbyqxmd.com/read/27444565/microrna-146a-in-human-and-experimental-ischemic-aki-cxcl8-dependent-mechanism-of-action
#20
Lucile Amrouche, Geoffroy Desbuissons, Marion Rabant, Virginia Sauvaget, Clément Nguyen, Aurélien Benon, Pauline Barre, Clémentine Rabaté, Xavier Lebreton, Morgan Gallazzini, Christophe Legendre, Fabiola Terzi, Dany Anglicheau
AKI leads to tubular injury and interstitial inflammation that must be controlled to avoid the development of fibrosis. We hypothesized that microRNAs are involved in the regulation of the balance between lesion formation and adaptive repair. We found that, under proinflammatory conditions, microRNA-146a (miR-146a) is transcriptionally upregulated by ligands of IL-1 receptor/Toll-like receptor family members via the activation of NF-κB in cultured renal proximal tubular cells. In vivo, more severe renal ischemia-reperfusion injury (IRI) associated with increased expression of miR-146a in both allografts and urine of human kidney transplant recipients, and unilateral IRI in mice induced miR-146a expression in injured kidneys...
July 21, 2016: Journal of the American Society of Nephrology: JASN
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