Indoxyl sulfate induces complex redox alterations in mesangial cells

Indoxyl sulfate is a protein metabolite that is concentrated in the serum of patients with chronic renal insufficiency. It also is a uremic toxin that has been implicated in the progression of chronic renal disease in rodent models. We have shown previously that mesangial cell redox status is related to activation of mitogen-activated protein kinases and cell proliferation, which are factors related to glomerular damage. We used three methods to examine the ability of indoxyl sulfate to alter mesangial cell redox as a possible mechanism for its toxicity. Indoxyl sulfate increases mesangial cell reduction rate in a concentration-dependent manner as demonstrated by redox microphysiometry. Alterations occurred at concentrations as low as 100 μM, with more marked alterations occurring at higher concentrations associated with human renal failure. We demonstrated that indoxyl sulfate induces the production of intracellular reactive oxygen species (ROS) in mesangial cells (EC50 = 550 μM) by using the ROS-sensitive fluorescent dye CM-DCF. ROS generation was only partially (∼50%) inhibited by the NADPH oxidase inhibitor diphenylene iodinium at low (≤300 μM) indoxyl sulfate concentrations. Diphenylene iodinium was without effect at higher concentrations of indoxyl sulfate. We also used electron paramagnetic spin resonance spectroscopy with extracellular and intracellular spin traps to show that indoxyl sulfate increases extracellular SOD-sensitive O2−· production and intracellular hydroxyl radical production that may derive from an initial O2−· burst. These results document that indoxyl sulfate, when applied to renal mesangial cells at pathological concentrations, induces rapid and complex changes in mesangial cell redox.

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Tumor necrosis factor alpha activates soluble guanylate cyclase in bovine glomerular mesangial cells via an L-arginine-dependent mechanism.

Journal of Experimental Medicine ◽

10.1084/jem.172.6.1843 ◽

1990 ◽

Vol 172 (6) ◽

pp. 1843-1852 ◽

Cited By ~ 80

Author(s):

P A Marsden ◽

B J Ballermann

Keyword(s):

Guanylate Cyclase ◽

Mesangial Cell ◽

Soluble Guanylate Cyclase ◽

Mesangial Cells ◽

Tnf Alpha ◽

Dependent Manner ◽

Glomerular Mesangial Cells ◽

Concentration Dependent Manner ◽

Necrosis Factor Alpha ◽

Factor Alpha

Endothelium-derived nitric oxide (NO) causes vasodilatation by activating soluble guanylate cyclase, and glomerular mesangial cells respond to NO with elevations of intracellular guanosine 3',5'-cyclic monophosphate (cGMP). We explored whether mesangial cells can be stimulated to produce NO and whether NO modulates mesangial cell function in an autocrine or paracrine fashion. Tumor necrosis factor alpha (TNF-alpha) raised mesangial cell cGMP levels in a time- and concentration-dependent manner (threshold dose 1 ng/ml, IC50 13.8 ng/ml, maximal response 100 ng/ml). TNF-alpha-induced increases in mesangial cGMP content were evident at 8 h and maximal at 18-24 h. The TNF-alpha-induced stimulation of mesangial cell cGMP production was abrogated by actinomycin D or cycloheximide suggesting dependence on new RNA or protein synthesis. Hemoglobin and methylene blue, both known to inhibit NO action, dramatically reduced TNF-alpha-induced mesangial cell cGMP production. Superoxide dismutase, known to potentiate NO action, augmented the TNF-alpha-induced effect. Ng-monomethyl-L-arginine (L-NMMA) decreased cGMP levels in TNF-alpha-treated, but not vehicle-treated mesangial cells in a concentration-dependent manner (IC50 53 microM). L-arginine had no effect on cGMP levels in control or TNF-alpha-treated mesangial cells but reversed L-NMMA-induced inhibition. Interleukin 1 beta and lipopolysaccharide (LPS), but not interferon gamma, also increased mesangial cell cGMP content. Transforming growth factor beta 1 blunted the mesangial cell response to TNF-alpha. TNF-alpha-induced L-arginine-dependent increases in cGMP were also evident in bovine renal artery vascular smooth muscle cells, COS-1 cells, and 1502 human fibroblasts. These findings suggest that TNF-alpha induces expression in mesangial cell of an enzyme(s) involved in the formation of L-arginine-derived NO. Moreover, the data indicate that NO acts in an autocrine and paracrine fashion to activate mesangial cell soluble guanylate cyclase. Cytokine-induced formation of NO in mesangial and vascular smooth muscle cells may be implicated in the pathogenesis of septic shock.

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Regulation of Rat Mesangial Cell Migration by Platelet-Derived Growth Factor, Angiotensin II, and Adrenomedullin

Journal of the American Society of Nephrology ◽

10.1681/asn.v10122495 ◽

1999 ◽

Vol 10 (12) ◽

pp. 2495-2502 ◽

Cited By ~ 6

Author(s):

MASAKAZU KOHNO ◽

KENICHI YASUNARI ◽

MIEKO MINAMI ◽

HIROAKI KANO ◽

KENSAKU MAEDA ◽

...

Keyword(s):

Cell Migration ◽

Angiotensin Ii ◽

Growth Factor ◽

Mesangial Cell ◽

Mesangial Cells ◽

Inhibitory Effect ◽

Platelet Derived Growth Factor ◽

Dependent Manner ◽

Glomerular Mesangial Cells ◽

Concentration Dependent Manner

Abstract. This study sought to determine whether platelet-derived growth factor (PDGF) and angiotensin II (AngII) stimulate migration of cultured rat glomerular mesangial cells. After finding that this was so, the effects of adrenomedullin (ADM) and cAMP-elevating agents on basal and stimulated mesangial cell migration were examined. Two isoforms of PDGF, AB and BB, stimulated migration in a concentration-dependent manner between 1 and 50 ng/ml, while the AA isoform lacked significant effect. AngII modestly but significantly stimulated migration in a concentration-dependent manner between 10-7 and 10-6 mol/L. Rat ADM significantly inhibited the PDGF BB- and AngII-stimulated migration in a concentration-dependent manner between 10-8 and 10-7 mol/L. Inhibition by rat ADM was accompanied by an increase in cellular cAMP. cAMP agonists or inducers such as 8-bromo cAMP, forskolin, and prostaglandin I2 also significantly reduced the stimulated migration. H 89, a protein kinase A (PKA) inhibitor, attenuated the inhibitory effect of ADM, and a calcitonin gene-related peptide (CGRP) receptor antagonist, human CGRP (8-37), abolished the inhibitory effects of rat ADM. These results suggest that PDGF AB and BB as well as AngII stimulate rat mesangial cell migration and that ADM can inhibit PDGF BB- and AngII-stimulated migration, at least in part through cAMP-dependent mechanisms likely to involve specific ADM receptors with which CGRP interacts. The adenylate cyclase/cAMP/PKA system may be involved in the migration-inhibitory effect of ADM in these cells.

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A mechanism by which atrial natriuretic factor mediates its glomerular actions

AJP Renal Physiology ◽

10.1152/ajprenal.1986.251.6.f1036 ◽

1986 ◽

Vol 251 (6) ◽

pp. F1036-F1042 ◽

Cited By ~ 4

Author(s):

R. G. Appel ◽

J. Wang ◽

M. S. Simonson ◽

M. J. Dunn

Keyword(s):

Mesangial Cell ◽

Mesangial Cells ◽

Dependent Manner ◽

Ang Ii ◽

Glomerular Mesangial Cells ◽

Sprague Dawley ◽

Cell Contraction ◽

Concentration Dependent Manner ◽

Cell Contractility

Differential in vivo glomerular effects of atriopeptin I (AP I) and atriopeptin III (AP III) were studied in parallel with in vitro physiological and biochemical parameters. In anesthetized Sprague-Dawley rats, AP III, but not AP I, significantly increased glomerular filtration rate. Image analysis microscopy was used to assess the effect of AP I and AP III on angiotensin II (ANG II)-induced contraction of cultured rat glomerular mesangial cells. AP III, but not AP I, inhibited ANG II-induced mesangial cell contraction in a concentration-dependent manner. Additional inhibitory agents included exogenous DBcGMP, 8-BrcGMP, Na nitroprusside, and DBcAMP. AP III stimulated mesangial cell cGMP with a lower threshold and greater maximum stimulation than AP I. Neither agent stimulated cAMP accumulation. Since mesangial cell contractility may regulate the glomerular capillary surface area, these results suggest that AP III partially mediates its glomerular effects through inhibition of ANG II-induced mesangial cell contraction. Whereas cGMP is not clearly implicated as the mediator of this effect, it appears that both cGMP and cAMP may regulate the state of mesangial cell contractility.

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Natriuretic peptides inhibit mesangial cell production of endothelin induced by arginine vasopressin

AJP Renal Physiology ◽

10.1152/ajprenal.1993.264.4.f678 ◽

1993 ◽

Vol 264 (4) ◽

pp. F678-F683 ◽

Cited By ~ 1

Author(s):

M. Kohno ◽

T. Horio ◽

M. Ikeda ◽

K. Yokokawa ◽

T. Fukui ◽

...

Keyword(s):

Arginine Vasopressin ◽

Natriuretic Peptides ◽

High Performance ◽

Culture Medium ◽

Mesangial Cell ◽

Mesangial Cells ◽

Dependent Manner ◽

Glomerular Mesangial Cells ◽

Concentration Dependent Manner ◽

Medium Level

The present study examined the effects of atrial, brain, and C-type natriuretic peptides (ANP, BNP, and CNP, respectively) on endothelin-1 (ET-1) secretion after stimulation with arginine vasopressin (AVP), using cultured rat glomerular mesangial cells. AVP stimulated immunoreactive (ir) ET-1 secretion in a concentration-dependent manner via a receptor-mediated process. Rat ANP-(1-28) and rat BNP-45 potently inhibited this stimulated secretion in a concentration-dependent manner. Inhibition by ANP and BNP of AVP-stimulated ET-1 secretion was paralleled by an increase in the medium level of guanosine 3',5'-cyclic monophosphate (cGMP). The addition of a cGMP analogue, 8-bromo-cGMP, reduced the stimulated ET-1 secretion. CNP was much less effective than rat ANP-(1-28) or rat BNP-45 with respect to inhibiting irET-1 secretion and increasing cGMP levels. High-performance liquid chromatography indicated that the major component of irET-1 in the culture medium corresponds to ET-1-(1-21). These findings indicate that AVP stimulates ET-1 secretion in cultured rat mesangial cells and that rat ANP and BNP inhibit this stimulated secretion, probably through a cGMP-dependent process.

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Blockade of the Effects of TGF-β1 on Mesangial Cells by Overexpression of Smad7

Journal of the American Society of Nephrology ◽

10.1681/asn.v134887 ◽

2002 ◽

Vol 13 (4) ◽

pp. 887-893 ◽

Cited By ~ 1

Author(s):

Ruihua Chen ◽

Cancan Huang ◽

Thomas A. Morinelli ◽

Maria Trojanowska ◽

Richard V. Paul

Keyword(s):

Transforming Growth Factor ◽

Mesangial Cell ◽

Mesangial Cells ◽

Glomerular Disease ◽

Luciferase Reporter ◽

Target Cells ◽

Dependent Manner ◽

Luciferase Reporter Gene ◽

Concentration Dependent Manner ◽

Tgf Β1

ABSTRACT. Smad7, a protein induced by transforming growth factor–β1 (TGF-β1) in many target cells, inhibits TGF-β1 signal transduction and is thought to mediate an intracellular negative feedback response that limits TGF-β1 effects. It is possible that overexpression of Smad7 could block specified effects of TGF-β1 on mesangial cells, a TGF-β target in glomerular disease. Smad7 mRNA was induced by TGF-β1 within 1 h in a concentration-dependent manner in a transformed mouse mesangial cell (MMC) line. Uptake of 14C-spermidine from the medium by MMC and the transcriptional activity of a segment of the human collagen pro-α2 type 1 chain (COL1A2) promoter fused to a luciferase reporter gene were used as indices of TGF-β1. Treatment with TGF-β1 increased 14C-spermidine uptake rate in a time-, concentration-, and temperature-dependent manner. For example, exposure to 1 ng/ml TGF-β1 for 15 h increased uptake approximately twofold, a response that was attenuated by cycloheximide. Transfection of Smad7 expression vector into MMC abrogated both TGF-β1-dependent stimulation of spermidine uptake and COL1A2 promoter activity. It is concluded that: (1) TGF-β1 induces Smad7 in MMC; (2) 14C-spermidine uptake is a convenient quantitative index of TGF-β1 effect in these cells; and (3) overexpression of Smad7 is a highly effective method of blocking at least some mesangial cell effects of TGF-β1 that may warrant evaluation in vivo in experimental glomerular disease.

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Faculty Opinions recommendation of Indoxyl sulfate, a representative uremic toxin, suppresses erythropoietin production in a HIF-dependent manner.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13477015.14857121 ◽

2012 ◽

Author(s):

Toshio Miyata

Keyword(s):

Indoxyl Sulfate ◽

Uremic Toxin ◽

Dependent Manner ◽

Erythropoietin Production

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Indoxyl-Sulfate-Induced Redox Imbalance in Chronic Kidney Disease

Antioxidants ◽

10.3390/antiox10060936 ◽

2021 ◽

Vol 10 (6) ◽

pp. 936

Author(s):

Chien-Lin Lu ◽

Cai-Mei Zheng ◽

Kuo-Cheng Lu ◽

Min-Tser Liao ◽

Kun-Lin Wu ◽

...

Keyword(s):

Oxidative Stress ◽

Chronic Kidney Disease ◽

Kidney Disease ◽

Muscle Wasting ◽

Mesangial Cells ◽

Target Organ Damage ◽

Organ Damage ◽

Indoxyl Sulfate ◽

Uremic Toxin ◽

Tubulointerstitial Injury

The accumulation of the uremic toxin indoxyl sulfate (IS) induces target organ damage in chronic kidney disease (CKD) patients, and causes complications including cardiovascular diseases, renal osteodystrophy, muscle wasting, and anemia. IS stimulates reactive oxygen species (ROS) production in CKD, which impairs glomerular filtration by a direct cytotoxic effect on the mesangial cells. IS further reduces antioxidant capacity in renal proximal tubular cells and contributes to tubulointerstitial injury. IS-induced ROS formation triggers the switching of vascular smooth muscular cells to the osteoblastic phenotype, which induces cardiovascular risk. Low-turnover bone disease seen in early CKD relies on the inhibitory effects of IS on osteoblast viability and differentiation, and osteoblastic signaling via the parathyroid hormone. Excessive ROS and inflammatory cytokine releases caused by IS directly inhibit myocyte growth in muscle wasting via myokines’ effects. Moreover, IS triggers eryptosis via ROS-mediated oxidative stress, and elevates hepcidin levels in order to prevent iron flux in circulation in renal anemia. Thus, IS-induced oxidative stress underlies the mechanisms in CKD-related complications. This review summarizes the underlying mechanisms of how IS mediates oxidative stress in the pathogenesis of CKD’s complications. Furthermore, we also discuss the potential role of oral AST-120 in attenuating IS-mediated oxidative stress after gastrointestinal adsorption of the IS precursor indole.

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Neurochemical Correlates of Brain Atrophy in Fibromyalgia Syndrome: A Magnetic Resonance Spectroscopy and Cortical Thickness Study

Brain Sciences ◽

10.3390/brainsci10060395 ◽

2020 ◽

Vol 10 (6) ◽

pp. 395

Author(s):

Paola Feraco ◽

Salvatore Nigro ◽

Luca Passamonti ◽

Alessandro Grecucci ◽

Maria Eugenia Caligiuri ◽

...

Keyword(s):

Magnetic Resonance ◽

Cortical Thickness ◽

Inferior Frontal Gyrus ◽

Pain Modulation ◽

Anterior Cingulate ◽

Resonance Spectroscopy ◽

Dependent Manner ◽

Cingulate Gyrus ◽

Concentration Dependent Manner ◽

The Right

(1) Background: Recently, a series of clinical neuroimaging studies on fibromyalgia (FM) have shown a reduction in cortical volume and abnormally high glutamate (Glu) and glutamate + glutamine (Glx) levels in regions associated with pain modulation. However, it remains unclear whether the volumetric decreases and increased Glu levels in FM are related each other. We hypothesized that higher Glu levels are related to decreases in cortical thickness (CT) and volume in FM patients. (2) Methods: Twelve females with FM and 12 matched healthy controls participated in a session of combined 3.0 Tesla structural magnetic resonance imaging (MRI) and single-voxel MR spectroscopy focused on the thalami and ventrolateral prefrontal cortices (VLPFC). The thickness of the cortical and subcortical gray matter structures and the Glu/Cr and Glx/Cr ratios were estimated. Statistics included an independent t-test and Spearman’s test. (3) Results: The Glu/Cr ratio of the left VLPFC was negatively related to the CT of the left inferior frontal gyrus (pars opercularis (p = 0.01; r = −0.75) and triangularis (p = 0.01; r = −0.70)). Moreover, the Glx/Cr ratio of the left VLPFC was negatively related to the CT of the left middle anterior cingulate gyrus (p = 0.003; r = −0.81). Significantly lower CTs in FM were detected in subparts of the cingulate gyrus on both sides and in the right inferior occipital gyrus (p < 0.001). (4) Conclusions: Our findings are in line with previous observations that high glutamate levels can be related, in a concentration-dependent manner, to the morphological atrophy described in FM patients.

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Sauchinone Protects Renal Mesangial Cell Dysfunction against Angiotensin II by Improving Renal Fibrosis and Inflammation

International Journal of Molecular Sciences ◽

10.3390/ijms21197003 ◽

2020 ◽

Vol 21 (19) ◽

pp. 7003

Author(s):

Jung Joo Yoon ◽

Hyeon Kyoung Lee ◽

Hye Yoom Kim ◽

Byung Hyuk Han ◽

Ho Sub Lee ◽

...

Keyword(s):

Cell Proliferation ◽

Diabetic Nephropathy ◽

Angiotensin Ii ◽

Mesangial Cell ◽

Mesangial Cells ◽

Tissue Growth ◽

Biologically Active ◽

Dependent Manner ◽

Saururus Chinensis ◽

Mesangial Cell Proliferation

Abnormal and excessive growth of mesangial cells is important in the pathophysiologic processes of diabetes-associated interstitial fibrosis and glomerulosclerosis, leading to diabetic nephropathy, which eventually turns into end-stage renal disease. Sauchinone, a biologically-active lignan isolated from aerial parts of Saururus chinensis, has anti-inflammatory and anti-viral activities effects on various cell types. However, there are no studies reporting the effects of sauchinone on diabetic nephropathy. The present study aims to investigate the role of sauchinone in mesangial cell proliferation and fibrosis induced by angiotensin II, as well as the underlying mechanisms of these processes. Human renal mesangial cells were induced by angiotensin II (AngII, 10 μM) in the presence or absence of sauchinone (0.1–1 μM) and incubated for 48 h. In this study, we found that AngII induced mesangial cell proliferation, while treatment with sauchinone inhibited the cell proliferation in a dose-dependent manner. Pre-treatment with sauchinone induced down-regulation of cyclins/CDKs and up-regulation of CDK inhibitor, p21, and p27kip1 expression. In addition, AngII-enhanced expression of fibrosis biomarkers such as fibronectin, collagen IV, and connective tissue growth factor (CTGF), which was markedly attenuated by sauchinone. Sauchinone also decreased AngII-induced TGF-β1 and Smad-2, Smad-3, and Smad-4 expression. This study further revealed that sauchinone ameliorated AngII-induced mesangial inflammation through disturbing activation of inflammatory factors, and NLRP3 inflammasome, which is composed of the NLRP3 protein, procaspase-1, and apoptosis-associated speck-like protein containing a CARD (ASC). Moreover, pretreatment of sauchinone inhibited NF-κB translocation and ROS production in AngII-exposed mesangial cells. These data suggest that sauchinone has a protective effect on renal proliferation, fibrosis and inflammation. Therefore, sauchinone might be a potential pharmacological agent in prevention of AngII-induced renal damage leading to diabetic nephropathy.

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