liu.seSearch for publications in DiVA
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
IGF-I/insulin hybrid receptors in human endothelial cells
Linköpings universitet, Institutionen för klinisk och experimentell medicin, Cellbiologi. Linköpings universitet, Hälsouniversitetet.
Linköpings universitet, Institutionen för klinisk och experimentell medicin, Cellbiologi. Linköpings universitet, Hälsouniversitetet.
Linköpings universitet, Institutionen för klinisk och experimentell medicin, Cellbiologi. Linköpings universitet, Hälsouniversitetet.
Department of Pathology, University of Washington School of Medicine, Seattle, USA.
Vise andre og tillknytning
2005 (engelsk)Inngår i: Molecular and Cellular Endocrinology, ISSN 0303-7207, E-ISSN 1872-8057, Vol. 229, nr 1-2, s. 31-37Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Vascular complications are common in diabetes. IGF-I receptors (IGF-IR) and insulin receptors (IR) in endothelial cells might respond to altered levels of IGF-I and insulin, resulting in altered endothelial function in diabetes. We therefore studied IGF-IR and IR gene expression, ligand binding, receptor protein, and phosphorylation in human umbilical vein endothelial cells (HUVEC). IGF-IR mRNA was more abundant than IRmRNAin freshly isolatedHUVEC(IGF-IR/IR ratio 7.1±1.5) and in culturedHUVEC(ratio 3.5±0.51). Accordingly, specific binding of 125I-IGF-I (0.64±0.25%) was higher than that of 125I-insulin (0.25±0.09%). Protein was detected for both  eceptors and IGF-I/insulin hybrid receptors. IGF-IR phosphorylation was stimulated by 10−10 to 10−8M IGF-I. IR were activated by 10−9 to 10−8M insulin and IGF-I. We conclude that HUVEC express more IGF-IR than IR, and also express hybrid receptors. Both IGF-I and insulin phosphorylate their own receptors but only IGF-I seems to phosphorylate hybrid receptors.

sted, utgiver, år, opplag, sider
2005. Vol. 229, nr 1-2, s. 31-37
Emneord [en]
IGF-I receptor; Insulin receptor; Insulin/IGF-I hybrid receptor; Endothelial cells; Human
HSV kategori
Identifikatorer
URN: urn:nbn:se:liu:diva-12553DOI: 10.1016/j.mce.2004.10.003ISI: 000226596400005OAI: oai:DiVA.org:liu-12553DiVA, id: diva2:1692
Tilgjengelig fra: 2008-09-15 Laget: 2008-09-15 Sist oppdatert: 2017-12-14
Inngår i avhandling
1. Expression and function of IGF-I and insulin receptors in human micro- and macrovascular cells
Åpne denne publikasjonen i ny fane eller vindu >>Expression and function of IGF-I and insulin receptors in human micro- and macrovascular cells
2008 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Insulin-like growth factor and insulin are phylogenetically closely related polypeptides and have large structural and biological similarities. Low circulating insulin-like growth factor-I (IGF-I), diabetes as well as insulin resistance have been implicated in the pathogenesis of cardiovascular disease, but the mechanisms involved are still not clear. Furthermore, little is known about direct effects of insulin-like growth factor-I (IGF-I) and insulin on human micro- and macrovascular cells.

In these studies we investigated the expression and function of insulin-like growth factor-I receptors (IGF-IR) and insulin receptors (IR) in human micro- and macrovascular endothelial cells and in human coronary artery smooth muscle cells.

Our results showed expression of both IGF-IR and IR in human dermal microvascular (HMVEC), aortic (HAEC) umbilical vein (HUVEC) and coronary artery (HCAEC) endothelial cells as well as in human coronary artery smooth muscle cells (HASMC). The gene expression of IGF-IR was several times more abundant than that of IR. Ligand binding studies confirmed that the IGF-IR was severalfold more abundant than the IR. It also demonstrated that insulin and glargine interacted with the IGF-IR with thousand- and hundredfold, respectively, less potency than IGF-I itself. The presence of IGF-IR and IR proteins and activation of their β-subunits was revealed by immunoprecipitation and Western blot analysis in human macrovascular endothelial cells and in coronary artery smooth muscle cells. At physiological concentrations (≤10-9 M) IGF-I and insulin activated their cognate receptors. The presence of hybrid IR/IGF-IR was shown through detection of the β-subunit for IGF-IR and IR on the same membrane by Western blot after immunoprecipitation with specific antibodies against either IGF-IR or IR, implying coprecipitation of the IGF-IR β-subunit and the IR β-subunit. The inability of physiological concentrations of insulin to phosphorylate IR β-subunit immunoprecipitated with IGF-IR antibodies and that IGF-I at physiological concentration activates the IR β-subunit is another evidence for the presence of the hybrid IR/IGF-IR. At physiological concentrations (≤10-9 M) IGF-I stimulated DNA synthesis and glucose incorporation into human coronary artery smooth muscle cells (HCASMC) and DNA synthesis in microvascular endothelial cells (HMVEC), but not in human macrovascular endothelial cells (HCAEC or HUVEC). No effect of insulin was found. Although physiological concentrations of insulin (≤10-9 M) were able to activate IR, insulin had no biological effects on the vascular cells studied. A possible explanation is that the insulin receptor signalling is too attenuated due to the presence of hybrid IR/IGF-IR and low number of IR expressed in the cells studied. Regarding the safety in the use of glargine, we show that glargine has 10-fold higher affinity for IGF-IR than human insulin. However, the glargine concentrations obtained in vivo during diabetes treatment is too low to affect the IGF-IR.

In conclusions our studies provide experimental evidence that human micro- and macrovascular endothelial and vascular smooth muscle cells express both IGF-IR and IR. Our in vitro data suggest that the cells studied are sensitive to IGF-I, but insensitive to insulin and this is due to the preponderance of IGF-IR and presence of hybrid IR/IGF-IR.

sted, utgiver, år, opplag, sider
Linköping: Linköping University Electronic Press, 2008. s. 89
Serie
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1045
HSV kategori
Identifikatorer
urn:nbn:se:liu:diva-12558 (URN)978-91-7393-972-0 (ISBN)
Disputas
2008-03-14, Berzeliussalen (hus 463, ingång 65), Hälsouniversitet, Campus US, Linköpings universitet, Linköping, 09:00 (svensk)
Opponent
Veileder
Tilgjengelig fra: 2008-09-15 Laget: 2008-09-15 Sist oppdatert: 2020-03-29bibliografisk kontrollert
2. IGF-I receptors, insulin receptors and insulin/IGF-I hybrid receptors in human endothelial cells: with special reference to diabetes
Åpne denne publikasjonen i ny fane eller vindu >>IGF-I receptors, insulin receptors and insulin/IGF-I hybrid receptors in human endothelial cells: with special reference to diabetes
2005 (engelsk)Licentiatavhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Patients with diabetes mellitus are known to develop vascular complications, which occur as macroangiopathy, atherosclerosis and mediasclerosis, as well as microangiopathy, e.g. retinopathy and nephropathy. The precise mechanisms causing these complications have not yet been elucidated. The microvascular complications are closely associated with the glycaemic control, which also is a risk factor for the diabetic macroangiopathy. The possible roles of insulin and the related peptide IGF-I, whose levels are affected by diabetes mellitus, are not clear. This study aims to characterise the presence and function of insulin and IGF-I receptors in human endothelial cells.

Two types of human endothelial cells were studied; human umbilical vein endothelial cells (HUVEC) and human coronary artery endothelial cells (HCAEC). The presence of insulin receptors and IGF-I receptors was studied at mRNA level by real-time PCR and at protein level by ligand binding and by Western blot analysis after immunoprecipitation. Receptor activation was determined as tyrosine phosphorylation.

Both HUVEC and HCAEC were found to express IGF-I receptors and insulin receptors at mRNA and protein levels. The amount of IGF-I receptor mRNA exceeded insulin receptor mRNA by 3.5 and 14-fold in HUVEC and HCAEC, respectively. In HUVEC, the higher expression of IGF-I receptor mRNA compared to insulin receptor mRNA was present in both freshly isolated and cultured cells. Ligand binding studies showed a higher specific binding of 125I-IGF-I than of 125I-insulin which also suggest the presence of more IGF-I receptors than insulin receptors. In HUVEC, the specific binding was 0.64 ± 0.25% (mean ± SEM) for 125I-IGF-I and 0.25 ± 0.092% for 125I-insulin. The EC50 for 125I-IGF-I displacement was 3.6 x 10-10 M for IGF-I vs. 8.25 x 10-8 M for insulin. The EC50 for 125I-insulin displacement was 2.6 x 10-10 M for insulin and 7.39 x 10-9 M for IGF-I. In HCAEC, the specific binding was 1.37 ± 0.09% and for insulin 0.17 ± 0.03%. The EC50 value for IGF-I displacement were 6.9 X 10-10 M for IGF-I, 8.7 X 10-6 M for insulin and 7.5 X 10-8 M for the insulin analogue glargine. Due to the very low specific binding of 125I-insulin, it was not possible to calculate the concentration needed to give half-maximal displacement, EC50, of 125I-labelled insulin. Both cell types expressed insulin/IGF-I hybrid receptors. Receptor phosphorylation studies showed that IGF-I receptor could be activated by 10-10 to 10-8 M IGF-I in both cell types. Insulin receptors were activated by 10-9 to 10-8 M insulin in HUVEC and HCAEC. IGF-I was able to activate insulin receptor phosphorylation at low concentrations, 10-9 to 10-8 M, which also is an indication of the presence of hybrid receptors.

In conclusion, two types of human endothelial cells, HUVEC and HCAEC, express more IGF-I receptors than insulin receptors, and they also express insulin/IGF-I hybrid receptors. IGF-I and insulin were found to phosphorylate their own receptor while IGF-I also seemed to be able to phosphorylate hybrid receptors. The results suggest an important role of the IGF-I receptor in human endothelial cells.

sted, utgiver, år, opplag, sider
Linköping: Linköpings universitet, 2005. s. 39
Serie
Linköping Studies in Health Sciences. Thesis, ISSN 1100-6013 ; 70
HSV kategori
Identifikatorer
urn:nbn:se:liu:diva-31179 (URN)16919 (Lokal ID)91-7393-857-3 (ISBN)16919 (Arkivnummer)16919 (OAI)
Tilgjengelig fra: 2009-10-09 Laget: 2009-10-09 Sist oppdatert: 2013-09-10

Open Access i DiVA

Fulltekst mangler i DiVA

Andre lenker

Forlagets fulltekstLink to Ph.D. thesis

Personposter BETA

Dekker Nitert, MarloesChisalitaa, Simona I.Arnqvist, Hans J.

Søk i DiVA

Av forfatter/redaktør
Dekker Nitert, MarloesChisalitaa, Simona I.Arnqvist, Hans J.
Av organisasjonen
I samme tidsskrift
Molecular and Cellular Endocrinology

Søk utenfor DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric

doi
urn-nbn
Totalt: 115 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf