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AP-1 proteins mediate hyperglycemia-induced activation of the human TGF-beta1 promoter in mesangial cells.
J Am Soc Nephrol. 2000 Nov; 11(11):2007-2016.JA

Abstract

Hyperglycemia-induced overproduction of the prosclerotic cytokine transforming growth factor-beta1 (TGF-beta1) has been implicated in the pathogenesis of diabetic nephropathy. Because high glucose and phorbol esters (PMA) increase TGF-beta1 mRNA levels in mesangial cells, this study was designed to characterize these effects on the human TGF-beta1 promoter activity. With the use of luciferase reporter gene constructs containing TGF-beta1 5'-flanking sequence (from -453 to +11 bp) transfected into mesangial cells, it was found that 30 mM glucose induced a nearly twofold increase in TGF-beta1 promoter activity after 24 h of incubation in human and porcine mesangial cells. Stimulation by PMA was more effective (2.3-fold). Mutagenesis in either one of the two or both activating protein-1 (AP-1) binding sites abolished the high glucose and the PMA effect. Furthermore, addition of the AP-1 inhibitor curcumin obliterated the glucose response. Corresponding experiments revealed that the transcription factor stimulating protein 1 was not involved in mediating the glucose effect. The high glucose-induced TGF-beta1 promoter activation was also prevented by inhibitors of protein kinase C and p38 mitogen-activated proteinkinase. Electrophoretic mobility shift assays with oligonucleotides containing one of the two AP-1 binding sites showed that glucose treatment markedly enhanced the binding activity of nuclear proteins of mesangial cells, particularly to box B. Supershift assays demonstrated that JunD and c-Fos were present in the protein-DNA complexes under control and hyperglycemic conditions. The functional and structural results show that glucose regulates human TGF-beta1 gene expression through two adjacent AP-1 binding sites and gives rise to the involvement of protein kinase C and p38 mitogen-activated protein kinase in hyperglycemia-induced TGF-beta1 gene expression.

Authors+Show Affiliations

Department of Internal Medicine, Division of Endocrinology, Metabolism and Pathobiochemistry, University of Tübingen, Tübingen, Germany.Institute of Pathology, University of Munich, Munich, Germany.Department of Internal Medicine, Division of Endocrinology, Metabolism and Pathobiochemistry, University of Tübingen, Tübingen, Germany.Medical Center Hospital Bergmannsheil, Bochum, Germany.Department of Internal Medicine, Division of Endocrinology, Metabolism and Pathobiochemistry, University of Tübingen, Tübingen, Germany.Department of Internal Medicine, Division of Endocrinology, Metabolism and Pathobiochemistry, University of Tübingen, Tübingen, Germany.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11053476

Citation

Weigert, Cora, et al. "AP-1 Proteins Mediate Hyperglycemia-induced Activation of the Human TGF-beta1 Promoter in Mesangial Cells." Journal of the American Society of Nephrology : JASN, vol. 11, no. 11, 2000, pp. 2007-2016.
Weigert C, Sauer U, Brodbeck K, et al. AP-1 proteins mediate hyperglycemia-induced activation of the human TGF-beta1 promoter in mesangial cells. J Am Soc Nephrol. 2000;11(11):2007-2016.
Weigert, C., Sauer, U., Brodbeck, K., Pfeiffer, A., Häring, H. U., & Schleicher, E. D. (2000). AP-1 proteins mediate hyperglycemia-induced activation of the human TGF-beta1 promoter in mesangial cells. Journal of the American Society of Nephrology : JASN, 11(11), 2007-2016. https://doi.org/10.1681/ASN.V11112007
Weigert C, et al. AP-1 Proteins Mediate Hyperglycemia-induced Activation of the Human TGF-beta1 Promoter in Mesangial Cells. J Am Soc Nephrol. 2000;11(11):2007-2016. PubMed PMID: 11053476.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - AP-1 proteins mediate hyperglycemia-induced activation of the human TGF-beta1 promoter in mesangial cells. AU - Weigert,Cora, AU - Sauer,Ulrich, AU - Brodbeck,Katrin, AU - Pfeiffer,Andreas, AU - Häring,Hans U, AU - Schleicher,Erwin D, PY - 2000/10/29/pubmed PY - 2001/2/28/medline PY - 2000/10/29/entrez SP - 2007 EP - 2016 JF - Journal of the American Society of Nephrology : JASN JO - J Am Soc Nephrol VL - 11 IS - 11 N2 - Hyperglycemia-induced overproduction of the prosclerotic cytokine transforming growth factor-beta1 (TGF-beta1) has been implicated in the pathogenesis of diabetic nephropathy. Because high glucose and phorbol esters (PMA) increase TGF-beta1 mRNA levels in mesangial cells, this study was designed to characterize these effects on the human TGF-beta1 promoter activity. With the use of luciferase reporter gene constructs containing TGF-beta1 5'-flanking sequence (from -453 to +11 bp) transfected into mesangial cells, it was found that 30 mM glucose induced a nearly twofold increase in TGF-beta1 promoter activity after 24 h of incubation in human and porcine mesangial cells. Stimulation by PMA was more effective (2.3-fold). Mutagenesis in either one of the two or both activating protein-1 (AP-1) binding sites abolished the high glucose and the PMA effect. Furthermore, addition of the AP-1 inhibitor curcumin obliterated the glucose response. Corresponding experiments revealed that the transcription factor stimulating protein 1 was not involved in mediating the glucose effect. The high glucose-induced TGF-beta1 promoter activation was also prevented by inhibitors of protein kinase C and p38 mitogen-activated proteinkinase. Electrophoretic mobility shift assays with oligonucleotides containing one of the two AP-1 binding sites showed that glucose treatment markedly enhanced the binding activity of nuclear proteins of mesangial cells, particularly to box B. Supershift assays demonstrated that JunD and c-Fos were present in the protein-DNA complexes under control and hyperglycemic conditions. The functional and structural results show that glucose regulates human TGF-beta1 gene expression through two adjacent AP-1 binding sites and gives rise to the involvement of protein kinase C and p38 mitogen-activated protein kinase in hyperglycemia-induced TGF-beta1 gene expression. SN - 1046-6673 UR - https://www.unboundmedicine.com/medline/citation/11053476/AP_1_proteins_mediate_hyperglycemia_induced_activation_of_the_human_TGF_beta1_promoter_in_mesangial_cells_ L2 - https://jasn.asnjournals.org/cgi/pmidlookup?view=long&pmid=11053476 DB - PRIME DP - Unbound Medicine ER -