Abstract
Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen that promotes angiogenesis, vasculogenesis, and increases vascular permeability. VEGF is expressed in renal tubular epithelial cells and urinary VEGF excretion is increased in various glomerular disorders. However, the mechanisms underlying expression of VEGF in renal tubular epithelial cells have not been fully elucidated. In the present study, we attempted to define a predominant regulator of VEGF expression using a cultured murine renal proximal tubular epithelial cell line (mProx24). VEGF protein concentration in the culture supernatant was measured by sandwich enzyme-linked immunosorbent assay. mProx24 constitutively produced VEGF at low level. Major isoforms expressed in this cell line were VEGF164 and VEGF120 determined by reverse transcription-polymerase chain reaction method. Among various stimuli including angiotensin II, transforming growth factor-beta1 (TGF-beta1), lipopolysaccharides, interleukin-1beta, interleukin-10 and interferon-gamma, only TGF-beta1 significantly increased the level of VEGF protein at 24 h in a dose-dependent manner. The steady-state mRNA level of VEGF was dose dependently increased by TGF-beta1 detected by Northern blotting. Treatment with neutralizing anti-TGF-beta1 antibody abolished TGF-beta1-induced VEGF expression by 70%. Inhibitors of protein kinase C (PKC), Ro-31-8220 and staurosporin, significantly suppressed TGF-beta1-induced VEGF protein expression. These results demonstrate the role of TGF-beta1 on the expression of VEGF in proximal tubular epithelial cells mediated potentially via PKC pathway. This regulatory mechanism may be associated with the progression of tubulointerstitial lesions in renal disorders.
| Original language | English |
|---|---|
| Journal | Nephron - Experimental Nephrology |
| Volume | 95 |
| Issue number | 2 |
| Publication status | Published - 2003 |
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Transforming growth factor-beta 1 induces vascular endothelial growth factor expression in murine proximal tubular epithelial cells. / Kitamura, Shinji; Maeshima, Yohei; Sugaya, Takeshi; Sugiyama, Hitoshi; Yamasaki, Yasushi; Makino, Hirofumi.
In: Nephron - Experimental Nephrology, Vol. 95, No. 2, 2003.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Transforming growth factor-beta 1 induces vascular endothelial growth factor expression in murine proximal tubular epithelial cells.
AU - Kitamura, Shinji
AU - Maeshima, Yohei
AU - Sugaya, Takeshi
AU - Sugiyama, Hitoshi
AU - Yamasaki, Yasushi
AU - Makino, Hirofumi
PY - 2003
Y1 - 2003
N2 - Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen that promotes angiogenesis, vasculogenesis, and increases vascular permeability. VEGF is expressed in renal tubular epithelial cells and urinary VEGF excretion is increased in various glomerular disorders. However, the mechanisms underlying expression of VEGF in renal tubular epithelial cells have not been fully elucidated. In the present study, we attempted to define a predominant regulator of VEGF expression using a cultured murine renal proximal tubular epithelial cell line (mProx24). VEGF protein concentration in the culture supernatant was measured by sandwich enzyme-linked immunosorbent assay. mProx24 constitutively produced VEGF at low level. Major isoforms expressed in this cell line were VEGF164 and VEGF120 determined by reverse transcription-polymerase chain reaction method. Among various stimuli including angiotensin II, transforming growth factor-beta1 (TGF-beta1), lipopolysaccharides, interleukin-1beta, interleukin-10 and interferon-gamma, only TGF-beta1 significantly increased the level of VEGF protein at 24 h in a dose-dependent manner. The steady-state mRNA level of VEGF was dose dependently increased by TGF-beta1 detected by Northern blotting. Treatment with neutralizing anti-TGF-beta1 antibody abolished TGF-beta1-induced VEGF expression by 70%. Inhibitors of protein kinase C (PKC), Ro-31-8220 and staurosporin, significantly suppressed TGF-beta1-induced VEGF protein expression. These results demonstrate the role of TGF-beta1 on the expression of VEGF in proximal tubular epithelial cells mediated potentially via PKC pathway. This regulatory mechanism may be associated with the progression of tubulointerstitial lesions in renal disorders.
AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen that promotes angiogenesis, vasculogenesis, and increases vascular permeability. VEGF is expressed in renal tubular epithelial cells and urinary VEGF excretion is increased in various glomerular disorders. However, the mechanisms underlying expression of VEGF in renal tubular epithelial cells have not been fully elucidated. In the present study, we attempted to define a predominant regulator of VEGF expression using a cultured murine renal proximal tubular epithelial cell line (mProx24). VEGF protein concentration in the culture supernatant was measured by sandwich enzyme-linked immunosorbent assay. mProx24 constitutively produced VEGF at low level. Major isoforms expressed in this cell line were VEGF164 and VEGF120 determined by reverse transcription-polymerase chain reaction method. Among various stimuli including angiotensin II, transforming growth factor-beta1 (TGF-beta1), lipopolysaccharides, interleukin-1beta, interleukin-10 and interferon-gamma, only TGF-beta1 significantly increased the level of VEGF protein at 24 h in a dose-dependent manner. The steady-state mRNA level of VEGF was dose dependently increased by TGF-beta1 detected by Northern blotting. Treatment with neutralizing anti-TGF-beta1 antibody abolished TGF-beta1-induced VEGF expression by 70%. Inhibitors of protein kinase C (PKC), Ro-31-8220 and staurosporin, significantly suppressed TGF-beta1-induced VEGF protein expression. These results demonstrate the role of TGF-beta1 on the expression of VEGF in proximal tubular epithelial cells mediated potentially via PKC pathway. This regulatory mechanism may be associated with the progression of tubulointerstitial lesions in renal disorders.
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UR - http://www.scopus.com/inward/citedby.url?scp=1542545613&partnerID=8YFLogxK
M3 - Article
C2 - 14610327
AN - SCOPUS:1542545613
VL - 95
JO - Experimental Nephrology
JF - Experimental Nephrology
SN - 0028-2766
IS - 2
ER -