Stimulation of TGF-β type II receptor by high glucose in mouse mesangial cells and in diabetic kidney

2000 ◽  
Vol 278 (5) ◽  
pp. F830-F838 ◽  
Author(s):  
Motohide Isono ◽  
András Mogyorósi ◽  
Dong Cheol Han ◽  
Brenda B. Hoffman ◽  
Fuad N. Ziyadeh
Keyword(s):  
High Glucose ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Kidney Cortex ◽  
Mrna Levels ◽  
Type Ii ◽  
Diabetic Kidney ◽  
Normal Glucose ◽  
Cells Cultured ◽  
Stimulation Of

Transforming growth factor-β (TGF-β) is important in the pathogenesis of diabetic nephropathy, but little is known about the regulation of the ligand-binding TGF-β type II signaling receptor (TβIIR). There were significant increases in TβIIR protein and mRNA levels in kidney cortex after 1–6 wk of streptozotocin-induced diabetes. Mouse mesangial cells cultured in high glucose demonstrated significantly increased TβIIR protein and mRNA levels compared with normal glucose. This effect was independent of stimulation of TGF-β bioactivity by high glucose. Consistent with transcriptional activation by high glucose, the half-life (∼4 h) of TβIIR mRNA was not affected by glucose concentration. Moreover, mouse mesangial cells transiently transfected with reporter constructs containing the first 47- or 274-bp promoter fragments of TβIIR demonstrated significantly increased reporter activity in high glucose. Cells grown in high glucose demonstrated increased responsiveness to a relatively small dose of exogenous TGF-β1 (0.5 ng/ml): [3H]proline incorporation and α1(IV) collagen mRNA were significantly greater in cells cultured in high than in normal glucose. Hence, the expression of TβIIR is increased in the diabetic kidney and in mesangial cells cultured in high glucose, primarily because of stimulation of gene transcription. TβIIR upregulation by high ambient glucose may contribute to the increased sensitivity of mesangial cells to the profibrogenic action of TGF-β1.

scholarly journals Extracellular Signal-Regulated Kinase Mediates Stimulation of TGF-β1 and Matrix by High Glucose in Mesangial Cells

2000 ◽  
Vol 11 (12) ◽  
pp. 2222-2230
Author(s):  
MOTOHIDE ISONO ◽  
M. CARMEN IGLESIAS-DE LA CRUZ ◽  
SHELDON CHEN ◽  
SOON WON HONG ◽  
FUAD N. ZIYADEH
Keyword(s):  
High Glucose ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Specific Inhibitor ◽  
Mrna Levels ◽  
Erk Activation ◽  
Extracellular Signal Regulated Kinase ◽  
Extracellular Signal ◽  
Tgf Β1 ◽  
Stimulation Of

Abstract. High ambient glucose exerts its injurious effects on renal cells through nonenzymatic and enzymatic pathways, including altered signal transduction and upregulation of the transforming growth factor-β (TGF-β) system. Extracellular signal-regulated kinase (ERK), a member of the mitogen-activated protein kinase (MAPK) cascade, is activated in mesangial cells cultured in high glucose and in glomeruli of diabetic rats. However, the biologic consequences of ERK activation in the kidney have not been investigated. To clarify the role of ERK activation, mouse mesangial cells were exposed to normal (5.5 mM) or high (25 mM) glucose with or without addition of PD98059, a specific inhibitor of MAPK/ERK kinase (MEK), an upstream kinase activator of ERK. Cells that were exposed to high glucose exhibited significant increases in ERK activity, TGF-β1 expression (total protein, mRNA levels, and promoter activity), [3H]-proline uptake, and α1(I) collagen and fibronectin mRNA levels. Treatment with PD98059 (up to 25 μM) significantly inhibited these parameters. In contrast, 25 μM PD98059 had no significant effect on any of the parameters measured in cells that were exposed to normal glucose. Overexpression of MAPK phosphatase CL 100 prevented TGF-β1 promoter activation by high glucose, confirming the involvement of the MEK-ERK pathway in response to high glucose. The conclusion is that activation of ERK in mesangial cells is responsible for high-glucose-induced stimulation of TGF-β1 and contributes to the increased extracellular matrix expression.

scholarly journals BKCa Mediates Dysfunction in High Glucose Induced Mesangial Cell Injury via TGF-β1/Smad2/3 Signaling Pathways

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Zhigui Wu ◽  
Wenxian Yin ◽  
Mengqi Sun ◽  
Yuankai Si ◽  
Xiaoxiao Wu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Kidney Disease ◽  
Signaling Pathways ◽  
Transforming Growth Factor Beta ◽  
High Glucose ◽  
Diabetic Kidney Disease ◽  
Transforming Growth Factor ◽  
Mesangial Cell ◽  
Mesangial Cells ◽  
Diabetic Kidney

Objective. To explore the role and mechanism of BKCa in diabetic kidney disease. Methods. Rat mesangial cells (MCs) HBZY-1 were cultured with high glucose to simulate the high-glucose environment of diabetic kidney disease in vivo. The effects of large conductance calcium-activated potassium channel (BKCa) on proliferation, migration, and apoptosis of HBZY-1 cells were observed. The contents of transforming growth factor beta 1 (TGF-β1), Smad2/3, collagen IV (Col IV), and fibronectin (FN) in the extracellular matrix were also observed. Results. High glucose significantly damaged HBZY-1 cells, which enhanced the ability of cell proliferation, migration, and apoptosis, and increased the secretion of Col IV and FN. Inhibition of BKCa and TGF-β1/Smad2/3 signaling pathways can inhibit the proliferation, migration, and apoptosis of HBZY-1 cells and suppress the secretion of Col IV and FN. The effect of excitation is the opposite. Conclusions. BKCa regulates mesangial cell proliferation, migration, apoptosis, and secretion of Col IV and FN and is associated with TGF-β1/Smad2/3 signaling pathway.

Increased decorin mRNA in diabetic mouse kidney and in mesangial and tubular cells cultured in high glucose

1998 ◽  
Vol 275 (5) ◽  
pp. F827-F832 ◽  
Author(s):  
Andras Mogyorosi ◽  
Fuad N. Ziyadeh
Keyword(s):  
High Glucose ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Core Protein ◽  
Tissue Injury ◽  
Streptozotocin Diabetes ◽  
Mouse Kidney ◽  
Kidney Cortex ◽  
Tubular Cells ◽  
Proximal Tubular

The core protein of the proteoglycan decorin binds and neutralizes transforming growth factor-β (TGF-β). Activation of TGF-β is crucial to tissue injury in diabetic nephropathy, but it is not currently known whether decorin plays a role in this disease. Mouse kidney cortex demonstrates more than a twofold increase in decorin mRNA after 1, 2, 3, and 6 wk of streptozotocin diabetes. Various mouse and rat renal cell types are studied in culture under normal or high-glucose conditions. Mouse glomerular mesangial and proximal tubular epithelial cells constitutively express decorin, and high glucose (450 mg/dl) increases decorin mRNA fourfold compared with 100 mg/dl glucose. Unlike rat mesangial cells, rat glomerular epithelial and endothelial cells do not constitutively express decorin, and no induction is observed in high glucose. When mouse mesangial and proximal tubular cells are exposed to TGF-β1 (1 ng/ml), decorin mRNA is significantly decreased. Our findings suggest that the increased decorin expression in the diabetic kidney may counteract the hypertrophic and prosclerotic effects of increased TGF-β levels and that a negative feedback loop may exist between decorin and TGF-β.

Abstract 612: Apelinergic System within the Kidney is Altered in Diabetic Mice

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Anastasia Kalea ◽  
Minghao Ye ◽  
Jan Wysocki ◽  
Ivy Hsieh ◽  
Daniel Batlle
Keyword(s):  
Angiotensin Ii ◽  
Real Time ◽  
Real Time Pcr ◽  
Mesangial Cells ◽  
Diabetic Mouse ◽  
Kidney Cortex ◽  
Mrna Levels ◽  
Diabetic Kidney ◽  
Glomerular Epithelial Cells ◽  
Rapid Signaling

Background: The Apelin receptor (APJ)-Apelin system has been implicated in cardiovascular function but little is known regarding diabetic kidney disease. Apelin’s effects appear related, in part, to an interaction with angiotensin II. Apelin, like angiotensin II, is a substrate for ACE2, which we previously showed to localize in tubular and glomerular epithelial cells (podocytes). Here we wanted to investigate the localization of this peptide within the glomerulus and the overall expression of apelin and its receptor APJ within normal and diabetic mouse kidney. METHODS: We employed RT real time PCR, western blot, immunostaining, tissue and cell confocal microscopy to study apelin and APJ in kidneys from female C57BLKS mice (8 wks of age) and in cultured immortalized podocytes. Mouse podocytes were stimulated with Pyr 1 Apelin-13 (100nM) for 0, 1, 5, 15 and 60min and tested for phospho-status of AKT, p70S6K and ERK. Kidneys from 8 weeks old db/db mice were also studied. Results: Both Apelin and APJ were abundantly present in the kidney being expressed in most glomerular tufts, proximal tubules, and arterioles. In glomeruli, apelin colocalized with nephrin, PECAM-1, and a-SMA indicating its presence in podocytes, endothelial cells, and mesangial cells. APJ, by contrast, did not colocalize with PECAM-1 but also colocalized with several podocyte markers. In the proximal tubule, the APJ receptor colocalized with ACE2. Apelin and APJ mRNA were also expressed in cultured mouse podocytes where apelin-13 induced AKT, p70S6K and ERK rapid signaling (at 15min). By RT real time PCR, kidney cortex from db/db mice (n=6) had reduced APJ (0.19±0.02 vs 0.62±0.08, p< 0.005) and pre-proapelin mRNA levels (0.12±0.08 vs 1.13±0.04, p< 0.001) as compared to db/m controls (n=6). Conclusions: Apelin and its receptor are widely expressed in the kidney, including podocytes where apelin-13 is involved in AKT, p70S6K and ERK signaling. Reduced apelinergic activity in the diabetic kidney could be possibly compensatory to the known overactivity of the RAS.

scholarly journals Inhibitor of myogenic differentiation family isoform a, a new positive regulator of fibronectin production by glomerular mesangial cells

2020 ◽  
Vol 318 (3) ◽  
pp. F673-F682
Author(s):  
Parisa Yazdizadeh Shotorbani ◽  
Sarika Chaudhari ◽  
Yu Tao ◽  
Leonidas Tsiokas ◽  
Rong Ma
Keyword(s):  
Diabetic Nephropathy ◽  
High Glucose ◽  
Protein Level ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Myogenic Differentiation ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Glomerular Mesangial Cells ◽  
Significant Difference

Overproduction of extracellular matrix proteins, including fibronectin by mesangial cells (MCs), contributes to diabetic nephropathy. Inhibitor of myogenic differentiation family isoform a (I-mfa) is a multifunctional cytosolic protein functioning as a transcriptional modulator or plasma channel protein regulator. However, its renal effects are unknown. The present study was conducted to determine whether I-mfa regulated fibronectin production by glomerular MCs. In human MCs, overexpression of I-mfa significantly increased fibronectin abundance. Silencing I-mfa significantly reduced the level of fibronectin mRNA and blunted transforming growth factor-β1-stimulated production of fibronectin. We further found that high glucose increased I-mfa protein content in a time course (≥48 h) and concentration (≥25 mM)-dependent manner. Although high glucose exposure increased I-mfa at the protein level, it did not significantly alter transcripts of I-mfa in MCs. Furthermore, the abundance of I-mfa protein was significantly increased in the renal cortex of rats with diabetic nephropathy. The I-mfa protein level was also elevated in the glomerulus of mice with diabetic kidney disease. However, there was no significant difference in glomerular I-mfa mRNA levels between mice with and without diabetic nephropathy. Moreover, H2O2 significantly increased I-mfa protein abundance in a dose-dependent manner in cultured human MCs. The antioxidants polyethylene glycol-catalase, ammonium pyrrolidithiocarbamate, and N-acetylcysteine significantly blocked the high glucose-induced increase of I-mfa protein. Taken together, our results suggest that I-mfa, increased by high glucose/diabetes through the production of reactive oxygen species, stimulates fibronectin production by MCs.

Increased extracellular matrix synthesis and mRNA in mesangial cells grown in high-glucose medium

1991 ◽  
Vol 260 (2) ◽  
pp. F185-F191 ◽  
Author(s):  
S. H. Ayo ◽  
R. A. Radnik ◽  
W. F. Glass ◽  
J. A. Garoni ◽  
E. R. Rampt ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Protein Synthesis ◽  
High Glucose ◽  
Mesangial Cells ◽  
Matrix Proteins ◽  
Cell Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Glomerular Mesangial Cells ◽  
Glomerular Mesangium

Nodular expansion of glomerular mesangium with increased amounts of extracellular matrix (ECM) material is pathognomic of diabetic nephropathy. The precise mechanisms involved in this accumulation are unknown. Recently, we reported using a solid-phase enzyme-linked immunosorbent assay (ELISA) technique that glomerular mesangial cells, the principal cell type residing in glomerular mesangium, accumulate 50–60% more fibronectin (FN), laminin (LM), and type IV collagen (T-IV) when cultured in medium containing high glucose (30 mM) (S. H. Ayo, R. A. Rodnik, J. Garoni, W. F. Glass II, and J. I. Kreiberg. Am. J. Pathol. 136: 1339-1348, 1990). ECM assembly is controlled by its rate of synthesis and degradation, as well as its binding and rate of incorporation into the ECM. To elucidate the mechanisms involved, pulse-chase experiments were designed to estimate ECM protein synthesis from the incorporation of Trans-35S [( 35S]methionine, [35S]cysteine) into immunoprecipitated FN, LM, and T-IV. mRNA levels were examined, and degradation rates were estimated from the disappearance of radioactivity from matrix proteins in mesangial cells previously incubated with Trans-35S. One week of growth in 30 mM glucose resulted in approximately 40–50% increase in the synthesis of all three matrix proteins compared with 10 mM glucose-grown cells. This was accompanied by a significant increase in the transcripts for all three matrix proteins (approximately twofold). The specific activity of the radiolabel in trichloroacetic acid-precipitable cell protein showed no difference between cells grown in 10 or 30 mM glucose, indicating that total protein synthesis was unchanged. After 1 wk, the rate of FN, LM, and T-IV collagen degradation was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Renal hypertrophy is associated with upregulation of TGF-beta 1 gene expression in diabetic BB rat and NOD mouse

1994 ◽  
Vol 267 (6) ◽  
pp. F1094-F1001 ◽  
Author(s):  
K. Sharma ◽  
F. N. Ziyadeh
Keyword(s):  
Transforming Growth Factor ◽  
Nod Mouse ◽  
Dependent Diabetes Mellitus ◽  
Bb Rat ◽  
Mrna Levels ◽  
Glomerular Mesangial Cells ◽  
Tgf Beta ◽  
Renal Hypertrophy ◽  
Diabetic Kidney ◽  
Gene And Protein Expression

Renal hypertrophy is an early feature of diabetes, and it may predispose the kidney to the eventual development of parenchymal dysfunction. Since we have previously demonstrated that short-term culture in high glucose concentration stimulates production of transforming growth factor-beta 1 (TGF-beta 1) in proximal tubular and glomerular mesangial cells, we postulated that this cytokine, which has potent regulatory effects on cellular growth and extracellular matrix production, is important in mediating diabetic renal disease. In this study we evaluated the gene and protein expression of TGF-beta 1 in the kidney of two rodent models of spontaneous insulin-dependent diabetes mellitus [the biobreeding (BB) rat and the nonobese diabetic (NOD) mouse]. In association with the appearance in both models of significant renal hypertrophy, TGF-beta 1 mRNA levels were increased threefold in the kidney of the diabetic BB rat after 3 days of diabetes and also threefold after 7–9 days in the NOD mouse. There was no increase in TGF-beta 1 transcripts in the livers of the diabetic animals, suggesting that this response is tissue specific. Immunohistochemical studies revealed that TGF-beta 1 protein is concordantly elevated in the cortical tubular cells of the diabetic kidney in both models. These results suggest that the stimulated expression of renal TGF-beta is an early manifestation of the involvement of the kidney by diabetes. Whether increased TGF-beta production in the diabetic kidney is a key promoter of diabetic renal manifestations (e.g., hypertrophy) deserves additional studies.

Hypoxia and high glucose cause exaggerated mesangial cell growth and collagen synthesis: role of osteopontin

2001 ◽  
Vol 280 (4) ◽  
pp. F667-F674 ◽  
Author(s):  
Chhinder P. Sodhi ◽  
Sarojini A. Phadke ◽  
Daniel Batlle ◽  
Atul Sahai
Keyword(s):  
Cell Growth ◽  
High Glucose ◽  
Collagen Synthesis ◽  
Type Iv Collagen ◽  
Mesangial Cell ◽  
Mesangial Cells ◽  
Neutralizing Antibody ◽  
Cell Number ◽  
Mrna Levels ◽  
Type Iv

The effect of hypoxia on the proliferation and collagen synthesis of cultured rat mesangial cells was examined under normal-glucose (NG, 5 mM) and high-glucose (HG, 25 mM)-media conditions. In addition, a role for osteopontin (OPN) in mediating these processes was assessed. Quiescent cultures were exposed to hypoxia (3% O2) and normoxia (18% O2) in a serum-free medium with NG or HG, and cell proliferation, collagen synthesis, and OPN expression were assessed. Cells exposed to hypoxia in NG medium resulted in significant increases in [3H]thymidine incorporation, cell number, and [3H]proline incorporation, respectively. HG incubations also produced significant stimulation of these parameters under normoxic conditions, which were markedly enhanced in cells exposed to hypoxia in HG medium. In addition, hypoxia and HG stimulated the mRNA levels of type IV collagen, and the combination of hypoxia and HG resulted in additive increases in type IV collagen expression. Hypoxia and HG also stimulated OPN mRNA and protein levels in an additive fashion. A neutralizing antibody to OPN or its β3-integrin receptor significantly blocked the effect of hypoxia and HG on proliferation and collagen synthesis. In conclusion, these results demonstrate for the first time that hypoxia in HG medium produces exaggerated mesangial cell growth and type IV collagen synthesis. In addition, OPN appears to play a role in mediating the accelerated mesangial cell growth and collagen synthesis found in a hyperglycemic and hypoxic environment.

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