Skip to main content
Log in

Regulation of VEGF and bFGF mRNA expression and other proliferative compounds in skeletal muscle cells

  • Published:
Angiogenesis Aims and scope Submit manuscript

Abstract

The role of muscle contraction, prostanoids, nitric oxide and adenosine in the regulation of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and endothelial cell proliferative compounds in skeletal muscle cell cultures was examined. VEGF and bFGF mRNA, protein release as well as the proliferative effect of extracellular medium was determined in non-stimulated and electro-stimulated rat and human skeletal muscle cells. In rat skeletal muscle cells these aspects were also determined after treatment with inhibitors and/or donors of nitric oxide (NO), prostanoids and adenosine. Electro-stimulation caused an elevation in the VEGF and bFGF mRNA levels of rat muscle cells by 33% and 43% (P < 0.05), respectively, and in human muscle cells VEGF mRNA was elevated by 24%. Medium from electro-stimulated human, but not rat muscle cells induced a 126% higher (P < 0.05) endothelial cell proliferation than medium from non-stimulated cells. Cyclooxygenase inhibition of rat muscle cells induced a 172% increase (P < 0.05) in VEGF mRNA and a 104% increase in the basal VEGF release. Treatment with the NO donor SNAP (0.5 μM) decreased (P < 0.05) VEGF and bFGF mRNA by 42 and 38%, respectively. Medium from SNAP treated muscle cells induced a 45% lower (P < 0.05) proliferation of endothelial cells than control medium. Adenosine enhanced the basal VEGF release from muscle cells by 75% compared to control. The present data demonstrate that contractile activity, NO, adenosine and products of cyclooxygenase regulate the expression of VEGF and bFGF mRNA in skeletal muscle cells and that contractile activity and NO regulate endothelial cell proliferative compounds in muscle extracellular fluid.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • P Andersen J Henriksson (1977) ArticleTitleCapillary supply of the quadriceps femoris muscle of man: Adaptive response to exercise J Physiol 270 IssueID3 677–90

    Google Scholar 

  • F Goto K Goto K Weindel J Folkman (1993) ArticleTitleSynergistic effects of vascular endothelial growth factor and basic fibroblast growth factor on the proliferation and cord formation of bovine capillary endothelial cells within collagen gels [see comments] Lab Invest 69 IssueID5 508–17

    Google Scholar 

  • DW Leung G Cachianes WJ Kuang et al. (1989) ArticleTitleVascular endothelial growth factor is a secreted angiogenic mitogen Science 246 IssueID4935 1306–09

    Google Scholar 

  • EC Breen EC Johnson H Wagner et al. (1996) ArticleTitleAngiogenic growth factor mRNA responses in muscle to a single bout of exercise J Appl Physiol 81 IssueID1 355–61

    Google Scholar 

  • L Hoffner JJ Nielsen H Langberg Y Hellsten (2003) ArticleTitleExercise but not prostanoids enhance levels of vascular endothelial growth factor and other proliferative agents in human skeletal muscle interstitium J Physiol 550 IssueIDPt 1 217–25

    Google Scholar 

  • KG Peters S Werner G Chen LT Williams (1992) ArticleTitleTwo FGF receptor genes are differentially expressed in epithelial and mesenchymal tissues during limb formation and organogenesis in the mouse Development 114 IssueID1 233–43

    Google Scholar 

  • N Ferrara K Houck L Jakeman DW Leung (1992) ArticleTitleMolecular and biological properties of the vascular endothelial growth factor family of proteins Endocr Rev 13 IssueID1 18–32

    Google Scholar 

  • S Kanno N Oda M Abe et al. (1999) ArticleTitleEstablishment of a simple and practical procedure applicable to therapeutic angiogenesis Circulation 99 IssueID20 2682–87

    Google Scholar 

  • U Frandsen J Bangsbo H Langberg et al. (2000) ArticleTitleInhibition of nitric oxide synthesis by systemic N(G)-monomethyl-<sc>L</sc>-arginine administration in humans: Effects on interstitial adenosine, prostacyclin and potassium concentrations in resting and contracting skeletal muscle J Vasc Res 37 IssueID4 297–302

    Google Scholar 

  • M Karamouzis I Karamouzis E Vamvakoudis et al. (2001) ArticleTitleThe response of␣muscle interstitial prostaglandin E(2)(PGE(2)), prostacyclin I(2)(PGI(2)) and thromboxane A(2)(TXA(2)) levels during incremental dynamic exercise in humans determined by in vivo microdialysis Prostaglandins Leukot Essent Fatty Acids 64 IssueID4–5 259–63

    Google Scholar 

  • L Hoffner JJ Nielsen H Langberg Y Hellsten (2003) ArticleTitleExercise but not prostanoids enhance levels of vascular endothelial growth factor and other proliferative agents in human skeletal muscle interstitium J Physiol 550 IssueIDPt 1 217–25

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–18

    Google Scholar 

  • MK Jones H Wang BM Peskar et al. (1999) ArticleTitleInhibition of angiogenesis by nonsteroidal anti-inflammatory drugs: Insight into mechanisms and implications for cancer growth and ulcer healing Nat Med 5 IssueID12 1418–23

    Google Scholar 

  • JF Murphy C Steele O Belton DJ Fitzgerald (2003) ArticleTitleInduction of cyclo-oxygenase-1 and −2 modulates angiogenic responses to engagement of alphavbeta3 Br J Haematol 121 IssueID1 157–64

    Google Scholar 

  • D BenEzra (1978) ArticleTitleNeovasculogenic ability of prostaglandins, growth factors, and synthetic chemoattractants Am J Ophthalmol 86 IssueID4 455–61

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–18

    Google Scholar 

  • MM Hoper NF Voelkel TO Bates et al. (1997) ArticleTitleProstaglandins induce vascular endothelial growth factor in a human monocytic cell line and rat lungs via cAMP Am J Respir Cell Mol Biol 17 IssueID6 748–56

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–18

    Google Scholar 

  • S Harada JA Nagy KA Sullivan et al. (1994) ArticleTitleInduction of vascular endothelial growth factor expression by prostaglandin E2 and E1 in osteoblasts J Clin Invest 93 IssueID6 2490–96

    Google Scholar 

  • SC Pearce O. MD Hudlicka (2000) ArticleTitleEffect of indomethacin on capillary growth and microvasculature in chronically stimulated rat skeletal muscles J Physiol 526 IssueID(Pt 2 435–43

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–18

    Google Scholar 

  • TP Gavin DA Spector H Wagner et al. (2000) ArticleTitleNitric oxide synthase inhibition attenuates the skeletal muscle VEGF mRNA response to exercise J Appl Physiol 88 IssueID4 1192–98

    Google Scholar 

  • A RayChaudhury H Frischer AB Malik (1996) ArticleTitleInhibition of endothelial cell proliferation and bFGF-induced phenotypic modulation by nitric oxide J Cell Biochem 63 IssueID2 125–34

    Google Scholar 

  • PC Lee MR Kibbe MJ Schuchert et al. (2000) ArticleTitleNitric oxide induces angiogenesis and upregulates alpha(v)beta(3) integrin expression on endothelial cells Microvasc Res 60 IssueID3 269–80

    Google Scholar 

  • E Hashimoto K Kage T Ogita et al. (1994) ArticleTitleAdenosine as an endogenous mediator of hypoxia for induction of vascular endothelial growth factor mRNA in U-937 cells Biochem Biophys Res Commun 204 IssueID1 318–24

    Google Scholar 

  • JW Gu AL Brady V Anand et al. (1999) ArticleTitleAdenosine upregulates VEGF expression in cultured myocardial vascular smooth muscle cells Am J Physiol 277 IssueID2 Pt 2 H595–602

    Google Scholar 

  • J Bergström (1962) ArticleTitleMuscle electrolytes in man Determined by neuron activation analysis on needle biopsy specimen A study on normal subjects, kidney patients and patients with chronic diarrhoea Scand J Clin Lab Invest 14 87–93

    Google Scholar 

  • Y Hellsten U Frandsen (1997) ArticleTitleAdenosine formation in contracting primary rat skeletal muscle cells and endothelial cells in culture J Physiol 504 IssueIDPt 3 695–704

    Google Scholar 

  • IH Jonsdottir P Schjerling K Ostrowski et al. (2000) ArticleTitleMuscle contractions induce interleukin-6 mRNA production in rat skeletal muscles J Physiol 528 IssueIDPt 1 157–63

    Google Scholar 

  • J Higginson H Wackerhage N Woods et al. (2002) ArticleTitleBlockades of mitogen-activated protein kinase and calcineurin both change fibre-type markers in skeletal muscle culture Pflugers Arch 445 IssueID3 437–43

    Google Scholar 

  • E Tischer R Mitchell T Hartman et al. (1991) ArticleTitleThe human gene for vascular endothelial growth factor Multiple protein forms are encoded through alternative exon splicing. J Biol Chem 266 IssueID18 11947–54

    Google Scholar 

  • CY Cheung M Singh MJ Ebaugh RA Brace (1995) ArticleTitleVascular endothelial growth factor gene expression in ovine placenta and fetal membranes Am J Obstet Gynecol 173 IssueID3 Pt 1 753–59

    Google Scholar 

  • T Gustafsson K Bodin C Sylven et al. (2001) ArticleTitleIncreased expression of VEGF following exercise training in patients with heart failure Eur J Clin Invest 31 IssueID4 362–66

    Google Scholar 

  • L Jensen J Bangsbo Y Hellsten (2004) ArticleTitleEffect of high intensity training on capillarization and presence of angiogenic factors in human skeletal muscle J Physiol 557 IssueIDPt 2 571–82

    Google Scholar 

  • S Kastner MC Elias AJ Rivera Z Yablonka-Reuveni (2000) ArticleTitleGene expression patterns of the fibroblast growth factors and their receptors during myogenesis of rat satellite cells J Histochem Cytochem 48 IssueID8 1079–96

    Google Scholar 

  • J Alterio Y Courtois J Robelin et al. (1990) ArticleTitleAcidic and basic fibroblast growth factor mRNAs are expressed by skeletal muscle satellite cells Biochem Biophys Res Commun 166 IssueID3 1205–12

    Google Scholar 

  • EC Breen EC Johnson H Wagner et al. (1996) ArticleTitleAngiogenic growth factor mRNA responses in muscle to a single bout of exercise J Appl Physiol 81 IssueID1 355–61

    Google Scholar 

  • T Gustafsson A Puntschart L Kaijser et al. (1999) ArticleTitleExercise-induced expression of angiogenesis-related transcription and growth factors in human skeletal muscle Am J Physiol 276 IssueID2 Pt 2 H679–85

    Google Scholar 

  • RS Richardson H Wagner SR Mudaliar et al. (1999) ArticleTitleHuman VEGF gene expression in skeletal muscle: Effect of acute normoxic and hypoxic exercise Am J Physiol 277 IssueID6 Pt 2 H2247–52

    Google Scholar 

  • TP Gavin PD Wagner (2001) ArticleTitleEffect of short-term exercise training on angiogenic growth factor gene responses in rats J Appl Physiol 90 IssueID4 1219–26

    Google Scholar 

  • EC Breen EC Johnson H Wagner et al. (1996) ArticleTitleAngiogenic growth factor mRNA responses in muscle to a single bout of exercise J Appl Physiol 81 IssueID1 355–61

    Google Scholar 

  • T Gustafsson A Puntschart Kaijser et al. (1999) ArticleTitleExercise-induced expression of angiogenesis-related transcription and growth factors in human skeletal muscle Am J Physiol 276 IssueID2 Pt 2 H679–85

    Google Scholar 

  • RS Richardson H Wagner SR Mudaliar et al. (1999) ArticleTitleHuman VEGF gene expression in skeletal muscle: Effect of acute normoxic and hypoxic exercise Am J Physiol 277 IssueID6 Pt 2 H2247–52

    Google Scholar 

  • TP Gavin PD Wagner (2001) ArticleTitleEffect of short-term exercise training on angiogenic growth factor gene responses in rats J Appl Physiol 90 IssueID4 1219–26

    Google Scholar 

  • Jensen L, Pilegaard H, Neufer PD, Hellsten Y Effect of acute exercise and Exercise training on VEGF splice variants in human Skeletal Muscle. Am J Physiol Regul Integr Comp Physiol 2004

  • S Kanno N Oda M Abe et al. (1999) ArticleTitleEstablishment of a simple and practical procedure applicable to therapeutic angiogenesis Circulation 99 IssueID20 2682–87

    Google Scholar 

  • EC Breen EC Johnson H Wagner et al. (1996) ArticleTitleAngiogenic growth factor mRNA responses in muscle to a single bout of exercise J Appl Physiol 81 IssueID1 355–61

    Google Scholar 

  • T Gustafsson A Puntschart L Kaijser et al. (1999) ArticleTitleExercise-induced expression of angiogenesis-related transcription and growth factors in human skeletal muscle Am J Physiol 276 IssueID2 Pt 2 H679–85

    Google Scholar 

  • RS Richardson H Wagner SR Mudaliar et al. (1999) ArticleTitleHuman VEGF gene expression in skeletal muscle: Effect of acute normoxic and hypoxic exercise Am J Physiol 277 IssueID6 Pt 2 H2247–52

    Google Scholar 

  • EC Breen EC Johnson H Wagner et al. (1996) ArticleTitleAngiogenic growth factor mRNA responses in muscle to a single bout of exercise J Appl Physiol 81 IssueID1 355–61

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–18

    Google Scholar 

  • MM Hoper NF Voelkel TO Bates et al. (1997) ArticleTitleProstaglandins induce vascular endothelial growth factor in a human monocytic cell line and rat lungs via cAMP Am J Respir Cell Mol Biol 17 IssueID6 748–56

    Google Scholar 

  • MK Jones H Wang BM Peskar et al. (1999) ArticleTitleInhibition of angiogenesis by nonsteroidal anti-inflammatory drugs: Insight into mechanisms and implications for cancer growth and ulcer healing Nat Med 5 IssueID12 1418–23

    Google Scholar 

  • AJ Knox L Corbett J Stocks et al. (2001) ArticleTitleHuman airway smooth muscle cells secrete vascular endothelial growth factor: Up-regulation by bradykinin via a protein kinase C and prostanoid-dependent mechanism FASEB J 15 IssueID13 2480–8

    Google Scholar 

  • Y Sakai K Fujita H Sakai K Mizuno (2001) ArticleTitleProstaglandin E2 regulates the expression of basic fibroblast growth factor messenger RNA in normal human fibroblasts Kobe J Med Sci 47 IssueID1 35–45

    Google Scholar 

  • R Pai IL Szabo BA Soreghan et al. (2001) ArticleTitlePGE(2) stimulates VEGF expression in endothelial cells via ERK2/JNK1 signaling pathways Biochem Biophys Res Commun 286 IssueID5 923–8

    Google Scholar 

  • JF Murphy C Steele O Belton DJ Fitzgerald (2003) ArticleTitleInduction of cyclooxygenase-1 and −2 modulates angiogenic responses to engagement of alphavbeta3 Br J Haematol 121 IssueID1 157–64

    Google Scholar 

  • D BenEzra (1978) ArticleTitleNeovasculogenic ability of prostaglandins, growth factors, and synthetic chemoattractants Am J Ophthalmol 86 IssueID4 455–61

    Google Scholar 

  • D Nie M Lamberti A Zacharek et al. (2000) ArticleTitleThromboxane A(2) regulation of endothelial cell migration, angiogenesis, and tumor metastasis Biochem Biophys Res Commun 267 IssueID1 245–51

    Google Scholar 

  • K Fukuo T Inoue S Morimoto et al. (1995) ArticleTitleNitric oxide mediates cytotoxicity and basic fibroblast growth factor release in cultured vascular smooth muscle cells. A possible mechanism of neovascularization in atherosclerotic plaques J Clin Invest 95 IssueID2 669–76

    Google Scholar 

  • J Dulak A Jozkowicz A Dembinska-Kiec et al. (2000) ArticleTitleNitric oxide induces the synthesis of vascular endothelial growth factor by rat vascular smooth muscle cells Arterioscler Thromb Vasc Biol 20 IssueID3 659–66

    Google Scholar 

  • LR Silveira L Pereira-Da-Silva et al. (2003) ArticleTitleFormation of hydrogen peroxide and nitric oxide in rat skeletal muscle cells during contractions Free Radic Biol Med 35 IssueID5 455–64

    Google Scholar 

  • TP Gavin DA Spector H Wagner et al. (2000) ArticleTitleNitric oxide synthase inhibition attenuates the skeletal muscle VEGF mRNA response to exercise J Appl Physiol 88 IssueID4 1192–98

    Google Scholar 

  • LR Silveira L Pereira-Da-Silva C Juel Y Hellsten (2003) ArticleTitleFormation of hydrogen peroxide and nitric oxide in rat skeletal muscle cells during contractions Free Radic Biol Med 35 IssueID5 455–64

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–8

    Google Scholar 

  • A RayChaudhury H Frischer AB Malik (1996) ArticleTitleInhibition of endothelial cell proliferation and bFGF-induced phenotypic modulation by nitric oxide J Cell Biochem 63 IssueID2 125–34

    Google Scholar 

  • H Benoit M Jordan H Wagner PD Wagner (1999) ArticleTitleEffect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angiogenic growth factor gene expression J Appl Physiol 86 IssueID5 1513–8

    Google Scholar 

  • J Lynge G Schulte N Nordsborg et al. (2003) ArticleTitleAdenosine A 2B receptors modulate cAMP levels and induce CREB but not ERK1/2 and p38 phosphorylation in rat skeletal muscle cells Biochem Biophys Res Commun 307 IssueID1 180–7

    Google Scholar 

  • L Hoffner JJ Nielsen H Langberg Y Hellsten (2003) ArticleTitleExercise but not prostanoids enhance levels of vascular endothelial growth factor and other proliferative agents in human skeletal muscle interstitium J Physiol 550 IssueIDPt 1 217–25

    Google Scholar 

  • L Jensen J Bangsbo Y Hellsten (2004) ArticleTitleEffect of high intensity training on capillarization and presence of angiogenic factors in human skeletal muscle J Physiol 557 IssueIDPt 2 571–82

    Google Scholar 

  • S Kanno N Oda M Abe et al. (1999) ArticleTitleEstablishment of a simple and practical procedure applicable to therapeutic angiogenesis Circulation 99 IssueID20 2682–7

    Google Scholar 

  • I Kosmidou A Xagorari C Roussos A Papapetropoulos (2001) ArticleTitleReactive oxygen species stimulate VEGF production from C(2)C(12) skeletal myotubes through a PI3K/Akt pathway Am J Physiol Lung Cell Mol Physiol 280 IssueID4 L585–92

    Google Scholar 

  • G Wu AP Mannam J Wu et al. (2003) ArticleTitleHypoxia induces myocyte-dependent COX-2 regulation in endothelial cells: Role of VEGF Am J Physiol Heart Circ Physiol 285 IssueID6 H2420–9

    Google Scholar 

  • W Risau (1997) ArticleTitleMechanisms of angiogenesis Nature 386 IssueID6626 671–74

    Google Scholar 

  • A Bikfalvi S Klein G Pintucci DB Rifkin (1997) ArticleTitleBiological roles of fibroblast growth factor-2 Endocr Rev 18 IssueID1 26–45

    Google Scholar 

  • JE Cartwright AP Johnstone GS Whitley (2000) ArticleTitleEndogenously produced nitric oxide inhibits endothelial cell growth as demonstrated using novel antisense cell lines Br J Pharmacol 131 IssueID1 131–7

    Google Scholar 

  • K Norrby (2000) ArticleTitleConstitutively synthesized nitric oxide is a physiological negative regulator of mammalian angiogenesis mediated by basic fibroblast growth factor Int J Exp Pathol 81 IssueID6 423–7

    Google Scholar 

  • PG Phillips LM Birnby A Narendran WL Milonovich (2001) ArticleTitleNitric oxide modulates capillary formation at the endothelial cell-tumor cell interface Am J Physiol Lung Cell Mol Physiol 281 IssueID1 L278–90

    Google Scholar 

  • A RayChaudhury H Frischer AB Malik (1996) ArticleTitleInhibition of endothelial cell proliferation and bFGF-induced phenotypic modulation by nitric oxide J Cell Biochem 63 IssueID2 125–34

    Google Scholar 

  • LR Silveira L Pereira-Da-Silva C Juel Y Hellsten (2003) ArticleTitleFormation of hydrogen peroxide and nitric oxide in rat skeletal muscle cells during contractions Free Radic Biol Med 35 IssueID5 455–64

    Google Scholar 

  • P Chomczynski N Sacchi (1987) ArticleTitleSingle-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction Analytical Biochemistry 162 156–59

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Y. Hellsten.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jensen, L., Schjerling, P. & Hellsten, Y. Regulation of VEGF and bFGF mRNA expression and other proliferative compounds in skeletal muscle cells. Angiogenesis 7, 255–267 (2004). https://doi.org/10.1007/s10456-004-4184-4

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10456-004-4184-4

Keywords

Navigation