Cyclical mechanical stretching increases the expression of vascular endothelial growth factor in rat vascular smooth muscle cells

Kou Gi Shyu, Mei Lung Chang, Bao Wei Wang, Peiliang Kuan, Hang Chang

研究成果: 雜誌貢獻文章

19 引文 (Scopus)

摘要

Background and Purpose: Mechanical forces have profound effects on vascular smooth muscle cells (VSMCs). The mechanism by which mechanical stimuli regulate vascular endothelial growth factor (VEGF) expression and regulation has yet to be elucidated. We investigated the effect of cyclical mechanical stretching on regulation of the VEGF gene in VSMCs. Materials and Methods: Cultured rat VSMCs grown on a flexible membrane base were stretched by applying a vacuum at 60 cycles/minute. VEGF concentration in the cultured media was determined by enzyme-linked immunoassay. VEGF gene expression was determined by Western blot and Northern blot. The location of VEGF in the VSMC was studied immunohistochemically. Chimeric constructs of the VEGF promoter were deleted and the promoter activity was determined by luciferase activity. Results: VEGF concentration increased by 21 to 32% as early as 10 minutes after stretching and remained at this level for up to 12 hours. The concentration of VEGF reached a maximum of 2.8-fold over that in control cells by 2 hours after stretching and declined slightly thereafter. The amount of VEGF mRNA in stretched cells increased as early as 1 hour after stretching, reached a maximum of 3.2-fold over the amount in control cells by 2 hours, and remained at this level for up to 6 hours after stretching. Immunohistochemical study confirmed increased VEGF expression in VSMCs after stretching. Stretched cells transfected with a Sac-Nhe fragment showed only 46% of the luciferase activity of unstretched control cells. However, stretched cells transfected with chimeric plasmids containing a Spe-Nhe fragment showed 2.8-fold luciferase activity over that in control cells. Conclusions: Cyclical mechanical stretching upregulates expression of the VEGF gene in VSMCs at the transcription level. The VEGF 5′-flanking region contains a negative stretch-response element located in the 0.4-kb Sac-Pst fragment and a positive stretch-response element located in the 0.6-kb Spe-Sac fragment.

原文英語
頁(從 - 到)741-747
頁數7
期刊Journal of the Formosan Medical Association = Taiwan yi zhi
100
發行號11
出版狀態已發佈 - 2001
對外發佈Yes

指紋

Vascular Smooth Muscle
Vascular Endothelial Growth Factor A
Smooth Muscle Myocytes
Luciferases
Response Elements
5' Flanking Region
Vacuum
Immunoenzyme Techniques
Northern Blotting
Genes
Plasmids
Up-Regulation
Western Blotting
Gene Expression
Messenger RNA

ASJC Scopus subject areas

  • Medicine(all)

引用此文

Cyclical mechanical stretching increases the expression of vascular endothelial growth factor in rat vascular smooth muscle cells. / Shyu, Kou Gi; Chang, Mei Lung; Wang, Bao Wei; Kuan, Peiliang; Chang, Hang.

於: Journal of the Formosan Medical Association = Taiwan yi zhi, 卷 100, 編號 11, 2001, p. 741-747.

研究成果: 雜誌貢獻文章

Shyu, Kou Gi ; Chang, Mei Lung ; Wang, Bao Wei ; Kuan, Peiliang ; Chang, Hang. / Cyclical mechanical stretching increases the expression of vascular endothelial growth factor in rat vascular smooth muscle cells. 於: Journal of the Formosan Medical Association = Taiwan yi zhi. 2001 ; 卷 100, 編號 11. 頁 741-747.
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title = "Cyclical mechanical stretching increases the expression of vascular endothelial growth factor in rat vascular smooth muscle cells",
abstract = "Background and Purpose: Mechanical forces have profound effects on vascular smooth muscle cells (VSMCs). The mechanism by which mechanical stimuli regulate vascular endothelial growth factor (VEGF) expression and regulation has yet to be elucidated. We investigated the effect of cyclical mechanical stretching on regulation of the VEGF gene in VSMCs. Materials and Methods: Cultured rat VSMCs grown on a flexible membrane base were stretched by applying a vacuum at 60 cycles/minute. VEGF concentration in the cultured media was determined by enzyme-linked immunoassay. VEGF gene expression was determined by Western blot and Northern blot. The location of VEGF in the VSMC was studied immunohistochemically. Chimeric constructs of the VEGF promoter were deleted and the promoter activity was determined by luciferase activity. Results: VEGF concentration increased by 21 to 32{\%} as early as 10 minutes after stretching and remained at this level for up to 12 hours. The concentration of VEGF reached a maximum of 2.8-fold over that in control cells by 2 hours after stretching and declined slightly thereafter. The amount of VEGF mRNA in stretched cells increased as early as 1 hour after stretching, reached a maximum of 3.2-fold over the amount in control cells by 2 hours, and remained at this level for up to 6 hours after stretching. Immunohistochemical study confirmed increased VEGF expression in VSMCs after stretching. Stretched cells transfected with a Sac-Nhe fragment showed only 46{\%} of the luciferase activity of unstretched control cells. However, stretched cells transfected with chimeric plasmids containing a Spe-Nhe fragment showed 2.8-fold luciferase activity over that in control cells. Conclusions: Cyclical mechanical stretching upregulates expression of the VEGF gene in VSMCs at the transcription level. The VEGF 5′-flanking region contains a negative stretch-response element located in the 0.4-kb Sac-Pst fragment and a positive stretch-response element located in the 0.6-kb Spe-Sac fragment.",
keywords = "Cell culture, Gene expression, Smooth muscle cell, Stretching, VEGF",
author = "Shyu, {Kou Gi} and Chang, {Mei Lung} and Wang, {Bao Wei} and Peiliang Kuan and Hang Chang",
year = "2001",
language = "English",
volume = "100",
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journal = "Journal of the Formosan Medical Association",
issn = "0929-6646",
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TY - JOUR

T1 - Cyclical mechanical stretching increases the expression of vascular endothelial growth factor in rat vascular smooth muscle cells

AU - Shyu, Kou Gi

AU - Chang, Mei Lung

AU - Wang, Bao Wei

AU - Kuan, Peiliang

AU - Chang, Hang

PY - 2001

Y1 - 2001

N2 - Background and Purpose: Mechanical forces have profound effects on vascular smooth muscle cells (VSMCs). The mechanism by which mechanical stimuli regulate vascular endothelial growth factor (VEGF) expression and regulation has yet to be elucidated. We investigated the effect of cyclical mechanical stretching on regulation of the VEGF gene in VSMCs. Materials and Methods: Cultured rat VSMCs grown on a flexible membrane base were stretched by applying a vacuum at 60 cycles/minute. VEGF concentration in the cultured media was determined by enzyme-linked immunoassay. VEGF gene expression was determined by Western blot and Northern blot. The location of VEGF in the VSMC was studied immunohistochemically. Chimeric constructs of the VEGF promoter were deleted and the promoter activity was determined by luciferase activity. Results: VEGF concentration increased by 21 to 32% as early as 10 minutes after stretching and remained at this level for up to 12 hours. The concentration of VEGF reached a maximum of 2.8-fold over that in control cells by 2 hours after stretching and declined slightly thereafter. The amount of VEGF mRNA in stretched cells increased as early as 1 hour after stretching, reached a maximum of 3.2-fold over the amount in control cells by 2 hours, and remained at this level for up to 6 hours after stretching. Immunohistochemical study confirmed increased VEGF expression in VSMCs after stretching. Stretched cells transfected with a Sac-Nhe fragment showed only 46% of the luciferase activity of unstretched control cells. However, stretched cells transfected with chimeric plasmids containing a Spe-Nhe fragment showed 2.8-fold luciferase activity over that in control cells. Conclusions: Cyclical mechanical stretching upregulates expression of the VEGF gene in VSMCs at the transcription level. The VEGF 5′-flanking region contains a negative stretch-response element located in the 0.4-kb Sac-Pst fragment and a positive stretch-response element located in the 0.6-kb Spe-Sac fragment.

AB - Background and Purpose: Mechanical forces have profound effects on vascular smooth muscle cells (VSMCs). The mechanism by which mechanical stimuli regulate vascular endothelial growth factor (VEGF) expression and regulation has yet to be elucidated. We investigated the effect of cyclical mechanical stretching on regulation of the VEGF gene in VSMCs. Materials and Methods: Cultured rat VSMCs grown on a flexible membrane base were stretched by applying a vacuum at 60 cycles/minute. VEGF concentration in the cultured media was determined by enzyme-linked immunoassay. VEGF gene expression was determined by Western blot and Northern blot. The location of VEGF in the VSMC was studied immunohistochemically. Chimeric constructs of the VEGF promoter were deleted and the promoter activity was determined by luciferase activity. Results: VEGF concentration increased by 21 to 32% as early as 10 minutes after stretching and remained at this level for up to 12 hours. The concentration of VEGF reached a maximum of 2.8-fold over that in control cells by 2 hours after stretching and declined slightly thereafter. The amount of VEGF mRNA in stretched cells increased as early as 1 hour after stretching, reached a maximum of 3.2-fold over the amount in control cells by 2 hours, and remained at this level for up to 6 hours after stretching. Immunohistochemical study confirmed increased VEGF expression in VSMCs after stretching. Stretched cells transfected with a Sac-Nhe fragment showed only 46% of the luciferase activity of unstretched control cells. However, stretched cells transfected with chimeric plasmids containing a Spe-Nhe fragment showed 2.8-fold luciferase activity over that in control cells. Conclusions: Cyclical mechanical stretching upregulates expression of the VEGF gene in VSMCs at the transcription level. The VEGF 5′-flanking region contains a negative stretch-response element located in the 0.4-kb Sac-Pst fragment and a positive stretch-response element located in the 0.6-kb Spe-Sac fragment.

KW - Cell culture

KW - Gene expression

KW - Smooth muscle cell

KW - Stretching

KW - VEGF

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C2 - 11802532

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JO - Journal of the Formosan Medical Association

JF - Journal of the Formosan Medical Association

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