Angiotensin AT1 receptor activation mediates high glucose-induced epithelial-mesenchymal transition in renal proximal tubular cells

Li Zhou; Hong Xue; Ping Yuan; Jun Ni; Chen Yu; Yu Huang; Li-Min Lu

doi:10.1111/j.1440-1681.2010.05421.x

Angiotensin AT₁ receptor activation mediates high glucose-induced epithelial-mesenchymal transition in renal proximal tubular cells

Li Zhou, Hong Xue, Ping Yuan, Jun Ni, Chen Yu, Yu Huang, Li-Min Lu

Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review

56 Citations (Scopus)

Abstract

1. Renal tubular epithelial cells can undergo epithelial to mesenchymal transition (EMT) under hyperglycaemic conditions, which is associated with renal interstitial fibrosis. Activation of the renin-angiotensin system (RAS) is involved in diabetic nephropathy. The present study investigated the positive role of angiotensin AT₁ receptors in high glucose-induced EMT in cultured tubular epithelial cells. 2. A rat kidney proximal tubular epithelial cell line (NRK-52E) was used in the present study. Levels of EMT makers, namely E-cadherin and vimentin, were estimated using fluorescence immunocytochemistry, mRNA levels of angiotensinogen (AGT), angiotensin-converting enzyme (ACE) and AT₁ receptors were determined by real-time polymerase chain reaction, protein levels of E-cadherin, vimentin, fibronectin, matrix metallopeptidase (MMP)-9 and phosphorylated extracellular signal-regulated kinase (ERK) 1/2 were analysed by western blotting and the concentrations of angiotensin (Ang) II and transforming growth factor (TGF)-β1 in the culture medium were determined by enzyme immunoassay and ELISA. 3. High glucose (30 mmol/L) induced EMT and increased the synthesis of fibronectin and MMP-9. Furthermore, high glucose increased AGT, ACE and AT₁ receptor mRNA levels, as well as AngII and TGF-β1 concentrations in the culture medium and ERK1/2 phosphorylation. Pretreatment of cells for 15 min with the AT₁ receptor antagonist losartan (10^-5 mol/L) attenuated high glucose-induced increases in TGF-β1 and ERK1/2 phosphorylation and reduced EMT, as well as the consequent synthesis of fibronectin and MMP-9. 4. The results of the present study suggest that the activated local RAS mediates high glucose-induced EMT. By activating AT₁ receptors and stimulating TGF-β1 synthesis, the elevated local RAS participates in high glucose-induced EMT and increased extracellular matrix secretion. © 2010 Blackwell Publishing Asia Pty Ltd.

Original language	English
Journal	Clinical and Experimental Pharmacology and Physiology
Volume	37
Issue number	9
DOIs	https://doi.org/10.1111/j.1440-1681.2010.05421.x
Publication status	Published - Sept 2010
Externally published	Yes

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Publication details (e.g. title, author(s), publication statuses and dates) are captured on an “AS IS” and “AS AVAILABLE” basis at the time of record harvesting from the data source. Suggestions for further amendments or supplementary information can be sent to [email protected].

Research Keywords

diabetic nephropathy
epithelial-mesenchymal transition
proximal tubular epithelial cell
renin-angiotensin system

UN SDGs

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@article{b38c820d80474c4a917d84b8b6bcc408,

title = "Angiotensin AT1 receptor activation mediates high glucose-induced epithelial-mesenchymal transition in renal proximal tubular cells",

abstract = "1. Renal tubular epithelial cells can undergo epithelial to mesenchymal transition (EMT) under hyperglycaemic conditions, which is associated with renal interstitial fibrosis. Activation of the renin-angiotensin system (RAS) is involved in diabetic nephropathy. The present study investigated the positive role of angiotensin AT1 receptors in high glucose-induced EMT in cultured tubular epithelial cells. 2. A rat kidney proximal tubular epithelial cell line (NRK-52E) was used in the present study. Levels of EMT makers, namely E-cadherin and vimentin, were estimated using fluorescence immunocytochemistry, mRNA levels of angiotensinogen (AGT), angiotensin-converting enzyme (ACE) and AT1 receptors were determined by real-time polymerase chain reaction, protein levels of E-cadherin, vimentin, fibronectin, matrix metallopeptidase (MMP)-9 and phosphorylated extracellular signal-regulated kinase (ERK) 1/2 were analysed by western blotting and the concentrations of angiotensin (Ang) II and transforming growth factor (TGF)-β1 in the culture medium were determined by enzyme immunoassay and ELISA. 3. High glucose (30 mmol/L) induced EMT and increased the synthesis of fibronectin and MMP-9. Furthermore, high glucose increased AGT, ACE and AT1 receptor mRNA levels, as well as AngII and TGF-β1 concentrations in the culture medium and ERK1/2 phosphorylation. Pretreatment of cells for 15 min with the AT1 receptor antagonist losartan (10-5 mol/L) attenuated high glucose-induced increases in TGF-β1 and ERK1/2 phosphorylation and reduced EMT, as well as the consequent synthesis of fibronectin and MMP-9. 4. The results of the present study suggest that the activated local RAS mediates high glucose-induced EMT. By activating AT1 receptors and stimulating TGF-β1 synthesis, the elevated local RAS participates in high glucose-induced EMT and increased extracellular matrix secretion. {\textcopyright} 2010 Blackwell Publishing Asia Pty Ltd.",

keywords = "diabetic nephropathy, epithelial-mesenchymal transition, proximal tubular epithelial cell, renin-angiotensin system",

author = "Li Zhou and Hong Xue and Ping Yuan and Jun Ni and Chen Yu and Yu Huang and Li-Min Lu",

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AU - Zhou, Li

AU - Xue, Hong

AU - Yuan, Ping

AU - Ni, Jun

AU - Yu, Chen

AU - Huang, Yu

AU - Lu, Li-Min

N1 - Publication details (e.g. title, author(s), publication statuses and dates) are captured on an “AS IS” and “AS AVAILABLE” basis at the time of record harvesting from the data source. Suggestions for further amendments or supplementary information can be sent to [email protected].

PY - 2010/9

Y1 - 2010/9

N2 - 1. Renal tubular epithelial cells can undergo epithelial to mesenchymal transition (EMT) under hyperglycaemic conditions, which is associated with renal interstitial fibrosis. Activation of the renin-angiotensin system (RAS) is involved in diabetic nephropathy. The present study investigated the positive role of angiotensin AT1 receptors in high glucose-induced EMT in cultured tubular epithelial cells. 2. A rat kidney proximal tubular epithelial cell line (NRK-52E) was used in the present study. Levels of EMT makers, namely E-cadherin and vimentin, were estimated using fluorescence immunocytochemistry, mRNA levels of angiotensinogen (AGT), angiotensin-converting enzyme (ACE) and AT1 receptors were determined by real-time polymerase chain reaction, protein levels of E-cadherin, vimentin, fibronectin, matrix metallopeptidase (MMP)-9 and phosphorylated extracellular signal-regulated kinase (ERK) 1/2 were analysed by western blotting and the concentrations of angiotensin (Ang) II and transforming growth factor (TGF)-β1 in the culture medium were determined by enzyme immunoassay and ELISA. 3. High glucose (30 mmol/L) induced EMT and increased the synthesis of fibronectin and MMP-9. Furthermore, high glucose increased AGT, ACE and AT1 receptor mRNA levels, as well as AngII and TGF-β1 concentrations in the culture medium and ERK1/2 phosphorylation. Pretreatment of cells for 15 min with the AT1 receptor antagonist losartan (10-5 mol/L) attenuated high glucose-induced increases in TGF-β1 and ERK1/2 phosphorylation and reduced EMT, as well as the consequent synthesis of fibronectin and MMP-9. 4. The results of the present study suggest that the activated local RAS mediates high glucose-induced EMT. By activating AT1 receptors and stimulating TGF-β1 synthesis, the elevated local RAS participates in high glucose-induced EMT and increased extracellular matrix secretion. © 2010 Blackwell Publishing Asia Pty Ltd.

AB - 1. Renal tubular epithelial cells can undergo epithelial to mesenchymal transition (EMT) under hyperglycaemic conditions, which is associated with renal interstitial fibrosis. Activation of the renin-angiotensin system (RAS) is involved in diabetic nephropathy. The present study investigated the positive role of angiotensin AT1 receptors in high glucose-induced EMT in cultured tubular epithelial cells. 2. A rat kidney proximal tubular epithelial cell line (NRK-52E) was used in the present study. Levels of EMT makers, namely E-cadherin and vimentin, were estimated using fluorescence immunocytochemistry, mRNA levels of angiotensinogen (AGT), angiotensin-converting enzyme (ACE) and AT1 receptors were determined by real-time polymerase chain reaction, protein levels of E-cadherin, vimentin, fibronectin, matrix metallopeptidase (MMP)-9 and phosphorylated extracellular signal-regulated kinase (ERK) 1/2 were analysed by western blotting and the concentrations of angiotensin (Ang) II and transforming growth factor (TGF)-β1 in the culture medium were determined by enzyme immunoassay and ELISA. 3. High glucose (30 mmol/L) induced EMT and increased the synthesis of fibronectin and MMP-9. Furthermore, high glucose increased AGT, ACE and AT1 receptor mRNA levels, as well as AngII and TGF-β1 concentrations in the culture medium and ERK1/2 phosphorylation. Pretreatment of cells for 15 min with the AT1 receptor antagonist losartan (10-5 mol/L) attenuated high glucose-induced increases in TGF-β1 and ERK1/2 phosphorylation and reduced EMT, as well as the consequent synthesis of fibronectin and MMP-9. 4. The results of the present study suggest that the activated local RAS mediates high glucose-induced EMT. By activating AT1 receptors and stimulating TGF-β1 synthesis, the elevated local RAS participates in high glucose-induced EMT and increased extracellular matrix secretion. © 2010 Blackwell Publishing Asia Pty Ltd.

KW - diabetic nephropathy

KW - epithelial-mesenchymal transition

KW - proximal tubular epithelial cell

KW - renin-angiotensin system

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DO - 10.1111/j.1440-1681.2010.05421.x

M3 - RGC 21 - Publication in refereed journal

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SN - 0305-1870

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JO - Clinical and Experimental Pharmacology and Physiology

JF - Clinical and Experimental Pharmacology and Physiology

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