Quantification of glomerular TGF-β1 mRNA in patients with diabetes mellitus

Iwano, Masayuki; Kubo, Atsushi; Nishino, Toshihiko; Sato, Hiroaki; Nishioka, Hisayuki; Akai, Yasuhiro; Kurioka, Hideyuki; Fujii, Yukihiko; Kanauchi, Masao; Sasaki, Hideo; Dohi, Kazuhiro

doi:10.1038/ki.1996.162

Cited by 176 publications

(113 citation statements)

References 31 publications

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“…Experiments in vitro have shown that exposure of renal cell cultures to high glucose concentrations results in elevated expression of TGF-β and ECM components [24,25]. Other researchers have confirmed these findings in experimental models for diabetic nephropathy and in patients with diabetes mellitus [26,27], suggesting that these factors play a role in the development of diabetic kidney disease [28]. Studies done in vitro and in vivo have shown that TGF-β elevates the synthesis of many ECM components, including collagen I, collagen III, collagen IV, fibronectin, laminin, and also of matrix glycoproteins such as osteopontin, osteonectin, tenascin, biglycan, and decorin [13,14,[29][30][31][32][33][34].…”

Section: Transforming Growth Factor-betamentioning

confidence: 82%

ECM homeostasis in renal diseases: a genomic approach

Eikmans

Baelde

Heer

et al. 2003

The Journal of Pathology

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Chronic renal disease is in general histologically accompanied by a vast amount of scar tissue, ie glomerulosclerosis and interstitial fibrosis. Scarring results from excessive accumulation of extracellular matrix (ECM) components, a process driven by a plethora of cytokines and growth factors. Studies in experimental renal disease which target these regulators using gene therapy limit or prevent the development of scarring. This review focuses specifically on the role of transforming growth factor-beta, platelet-derived growth factor, connective tissue growth factor, hepatocyte growth factor, and epidermal growth factor. The results obtained in animal models hold promise for molecular intervention strategies in human renal disease. Microarray technology allows large-scale gene expression profiling in kidney tissue to identify common molecular pathways in a step towards discovery of new drug targets. Molecular techniques are expected to be used for diagnostic and prognostic purposes in nephrological practice to supplement renal biopsy. Several studies already show that molecular techniques might be of use in routine diagnostic practice. Improvement of diagnosis and prediction of outcome in renal patients might lead to more efficient and earlier therapeutic intervention.

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Section: Transforming Growth Factor-betamentioning

confidence: 82%

ECM homeostasis in renal diseases: a genomic approach

Eikmans

Baelde

Heer

et al. 2003

The Journal of Pathology

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“…In fact, the in vitro findings of TGF-␤ induction by high glucose concentrations are accompanied by in vivo studies demonstrating increased TGF-␤ expression during hyperglycemia in animal models of diabetes. 1 Iwano et al 44 observed Less information is available on TGF-␤ receptor protein expression. 45 We observed increased expression of both a 105-kDa and a 65-kDa TGF-␤ receptor after exposure to glucose.…”

Section: Discussionmentioning

confidence: 99%

Glucose-Induced TGF-β1 and TGF-β Receptor-1 Expression in Vascular Smooth Muscle Cells Is Mediated by Protein Kinase C-α

Lindschau

Quass²,

Menne³

et al. 2003

Hypertension

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Abstract-Sclerosis and increased matrix expression in diabetes are mediated by glucose-induced transforming growth factor (TGF)-␤1 expression. The intracellular effects of high glucose occur at least in part by way of protein kinase C (PKC). We previously described a role for PKC-␣ in glucose-induced permeability. We now investigated the hypothesis that glucose-induced expression of TGF-␤1 and its receptors (TGF-␤-R1 and -R2) are mediated by activation of this PKC isoform. TGF-␤1 and TGF-␤-R expressions were determined in vascular smooth muscle cells (VSMCs) by immunocytochemistry and Western blotting. PKC isoforms were assessed by confocal microscopy. PKC isoforms were inhibited with antisense oligodeoxynucleotides. PKC-␣ was upregulated by overexpression or microinjection. High glucose (20 mmol/L) increased VSMC TGF-␤1 and TGF-␤-R1 expression but not TGF-␤-R2 expression. PKC inhibitors and specific PKC-␣ downregulation by antisense treatment prevented this effect, whereas antisense treatment against PKC-␤, -⑀, and -had no influence. PKC-␣ overexpression increased TGF-␤1 and TGF-␤-R1 expression but not TGF-␤-R2 expression. PKC-␣ microinjection into individual VSMCs also increased TGF-␤1 and TGF-␤-R immunofluorescence. Last, VSMCs from PKC-␣-deficient mice did not respond to high glucose compared with VSMCs from wild-type mice. We propose that high glucose-induced TGF-␤1 and TGF-␤-R1 expression is mediated by PKC-␣. Our findings suggest an autocrine feedback mechanism and a possible role for PKC-␣ in diabetic vascular disease. Key Words: glucose Ⅲ growth substances Ⅲ protein kinases Ⅲ diabetes Ⅲ muscle, vascular, smooth Ⅲ atherosclerosis R enal hypertrophy, basement membrane thickening, and extracellular matrix accumulation characterize diabetic nephropathy. 1 Several cytokines with fibrogenic potential contribute to this, particularly transforming growth factor (TGF)-␤1. 2 TGF-␤1 increases matrix synthesis, inhibits matrix degradation, upregulates adhesion molecules, and enhances chemoattraction. 3,4 The possible importance of TGF-␤1 in diabetic nephropathy has been demonstrated in vitro by glucose-induced upregulation of this cytokine in different cell types. 5-13 Increased TGF-␤1 expression has also been demonstrated in diabetic animal models and human patients. 14,15 TGF-␤ binds specific serine/threonine kinase type I and type II receptors. The receptors form heteromeric cell surface complexes with TGF-␤. 16,17 The type II receptor determines ligand specificity, whereas the type I receptor interacts with the type II receptor and might not have a specific ligand. The type II receptor, likely in conjunction with the type I receptor, is probably required for the antiproliferative effect, whereas the type I receptor is the probable mediator of extracellular gene expression. 16,17 High glucose upregulates TGF-␤1 receptors in vitro and in vivo. 18,19 Hyperglycemia might mediate the effect of protein kinase C (PKC) activation. 20 -24 PKC consists of several isoforms with distinct cellular functions and different ex...

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“…TGFβ1 mRNA expression was assessed with competitive RT-PCR using the primers 5′-AG-CGACTCGCCAGAGTGGTTATCTT-3′ and 5′-TTATGCTG-GTTGTACAGGGCCAGGA-3′ for TGFβ1 [20], and the primers 5′-CACCCACACTGTGCCCATCTA-3′ and 5′-ACTCGT-CATACTCCTGCTTGC-3′ [20] for the housekeeping gene β-actin which respectively give a 473 bp and 617 bp product. Each amplified product was confirmed by restricting endonuclease digestion with BamHI for TGFβ1 and BanI for β-actin.…”

Section: Generation Of Defective Adenovirus Expressing Gfat Cdnamentioning

confidence: 99%

P38 mitogen-activated protein kinase mediates hexosamine-induced TGFβ1 mRNA expression in human mesangial cells

et al. 2003

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Aims/hypothesis. The hexosamine pathway has been implicated in the induction of TGFβ1 expression and in the pathophysiology of diabetic glomerulopathy. Glucose-induced TGFβ1 expression is mediated by p38 mitogen-activated-protein-kinase (p38-MAPK) and this kinase is activated in the diabetic glomeruli. We examined whether the p38-MAPK is implicated in hexosamine-induced TGFβ1 mRNA expression in human mesangial cells. Methods. The products of the hexosamine biosynthetic pathway were increased by the addition of glucosamine or by the overexpression of the rate-limiting enzyme of the hexosamine pathway, glutamine: fructose-6-phosphate amidotransferase (GFAT).Results. Glucosamine addition resulted in cell death. UDP-N-Acetylglucosamine, one of the major hexosamine end-products, was increased in normal (7 mmol/l) and high (25 mmol/l) glucose conditions in GFAT-transfected cells compared to control transfected cells by twofold and 1.7-fold respectively (p≤0.04) and this was accompanied by a 1.6-and 2.3-fold increase (p≤0.02) in TGFβ1 mRNA expression. Addition of the GFAT inhibitor azaserine (10 µmol/l) prevented the induction of TGFβ1 in GFAT transfected cells. GFAT overexpression induced an increase in p38-MAPK activation after 6 and 12 h incubation in normal glucose, and this was prevented by the GFAT inhibitor azaserine. Furthermore, high glucose enhanced p38-MAPK activation in GFAT tranfected cells (p≤0.04). P38-MAPK inhibition using SB202190 (1 µmol/l) reduced hexosamine-induced TGFβ1 expression in normal and high glucose. The activation of the p38-MAPK was dependent on protein kinase-C. Conclusion/interpretation. Overexpression of GFAT increases hexosamine accumulation which mediates TGFβ1 expression via a protein kinase-C and p38-MAPK dependent mechanism. Increased glucose concentrations magnify these effects. [Diabetologia (2003) 46:531-537]

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Quantification of glomerular TGF-β1 mRNA in patients with diabetes mellitus

Cited by 176 publications

References 31 publications

ECM homeostasis in renal diseases: a genomic approach

ECM homeostasis in renal diseases: a genomic approach

Glucose-Induced TGF-β1 and TGF-β Receptor-1 Expression in Vascular Smooth Muscle Cells Is Mediated by Protein Kinase C-α

P38 mitogen-activated protein kinase mediates hexosamine-induced TGFβ1 mRNA expression in human mesangial cells

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