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Journal of Renin-Angiotensin-Aldosterone System
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Molecular mechanism(s) of insulin action on the expression of the angiotensinogen gene in kidney proximal tubular cells

Xiao-Hua Wu

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

Xing Chen

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

Shao-Ling Zhang

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

Li Pang

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

Catherine To

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

Tian-Tian Wang

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

Thomas C Hohman

Ayerst-Wyeth Research, Cardiovascular/ Metabolic Diseases, CN 8000, Princeton, NJ 08543-8000, USA

Janos G Filep

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4

John SD Chan

University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, 5415 boul. De l'Assomption, Montreal, Quebec, Canada, H1T 2M4, jchan{at}hmr.qc.ca

To investigate the molecular mechanism(s) of insulin action on the expression of the angiotensinogen (ANG) gene in kidney proximal tubular cells, we constructed a fusion gene, pOGH (hANG N-1064/+27), containing the 5'-flanking regulatory sequence of the human ANG gene fused with the human growth hormone (hGH) gene as a reporter and stably integrated the fusion gene into the opossum kidney (OK) cell genomes. The level of expression of pOGH (hANG N-1064/+27) was quantified by the amount of immunoreactive hGH secreted into the medium. The addition of a high level of D(+)-glucose (25 mM) or phorbol 12-myristate 13-acetate (PMA, 10-7 M) stimulated the expression of the fusion gene in OK cells. The stimulatory effect of glucose (25 mM) was blocked by insulin and tolrestat (an inhibitor of aldose reductase). Tolrestat also inhibited the increase of cellular DAG and PKC activity stimulated by 25 mM glucose. While insulin did not affect the cellular DAG and PKC activity, it did block the stimulatory effect of high glucose (25 mM) and PMA on the expression of the fusion gene. Finally, PD98059 (an inhibitor of mitogen-activated protein kinase kinase (MEK)) enhanced the stimulatory effect of high levels of glucose and blocked the inhibitory effect of insulin on the expression of the fusion gene as well as on the phosphorylation of MEK and mitogen-activated protein kinase (MAPK). In contrast, Wortmannin (an inhibitor of phosphatidylinositol-3-kinase) did not block the inhibitory effect of insulin on the ANG gene expression. These studies demonstrate that the action of insulin, blocking the stimulatory effect of a high level of D(+)-glucose (25 mM) on the ANG gene expression is mediated, at least in part, via the 5'flanking region of the ANG gene and MAPK signal transduction pathway.

Key Words: renin-angiotensin system • insulin • kidney

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Journal of Renin-Angiotensin-Aldosterone System, Vol. 1, No. 2, 166-174 (2000)
DOI: 10.3317/jraas.2000.021
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