A new model of primary human adipocytes reveals reduced early insulin signalling in type 2 diabetes
Standard
A new model of primary human adipocytes reveals reduced early insulin signalling in type 2 diabetes. / Algenstaedt, P; Rosenblatt, N; Kolb, I; Krützelmann, A; Schwarzloh, B; Böttcher, A; Wiesner, L; Greten, H; Hansen-Algenstaedt, N.
in: HORM METAB RES, Jahrgang 36, Nr. 8, 01.08.2004, S. 531-7.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - A new model of primary human adipocytes reveals reduced early insulin signalling in type 2 diabetes
AU - Algenstaedt, P
AU - Rosenblatt, N
AU - Kolb, I
AU - Krützelmann, A
AU - Schwarzloh, B
AU - Böttcher, A
AU - Wiesner, L
AU - Greten, H
AU - Hansen-Algenstaedt, N
PY - 2004/8/1
Y1 - 2004/8/1
N2 - The aim of this study was to establish a diabetic model of primary human adipocytes for investigating potential defects in early insulin signalling. Specimens of human subcutaneous adipose tissue were obtained during orthopaedic surgical procedures. Preadipocytes were isolated and differentiated to adipocytes. Western blot analysis and immunoprecipitation were performed to determine protein content of IRS-1, IRS-2, p85, phosphorylation of IRS-1, IRS-2, Akt and MAPK as well as association between p85 and IRS-1/IRS-2. In addition to short-term insulin stimulation, the effect of hyperinsulinaemia was investigated by treating cells with insulin over a period of 36 hours. We found a significantly reduced basal expression of IRS-1 (54 +/- 15%) in adipocytes from type 2 diabetic subjects compared to controls with a further significant reduction in expression after long-term treatment (30 +/- 12%) compared to short-term treatment. IRS-2 expression also showed a significant reduction under hyperinsulinaemic conditions (20 +/- 2%) in diabetics vs. controls. Furthermore, long-term treatment with insulin in diabetic adipocytes led to a significant reduction in the phosphorylation of IRS-1(68 +/- 11%), IRS-2 (82 +/- 11%), Akt (42 +/- 2%), and MAPK (92 +/- 12%) and in the subsequent association between p85 to IRS-1 and IRS-2 (100 +/- 16% and 96 +/- 12%) in comparison to controls. Investigating glucose uptake diabetic adipocytes revealed a significant reduction of 90 +/- 2%. In this study, we were able to establish a new diabetic model of primary human adipocytes. A defect in early insulin signalling in type 2 diabetic patients under hyperinsulinaemic conditions was determined. These results might help to give further insights in early insulin action; additionally, this human model represents a useful target for the study of new therapeutic approaches.
AB - The aim of this study was to establish a diabetic model of primary human adipocytes for investigating potential defects in early insulin signalling. Specimens of human subcutaneous adipose tissue were obtained during orthopaedic surgical procedures. Preadipocytes were isolated and differentiated to adipocytes. Western blot analysis and immunoprecipitation were performed to determine protein content of IRS-1, IRS-2, p85, phosphorylation of IRS-1, IRS-2, Akt and MAPK as well as association between p85 and IRS-1/IRS-2. In addition to short-term insulin stimulation, the effect of hyperinsulinaemia was investigated by treating cells with insulin over a period of 36 hours. We found a significantly reduced basal expression of IRS-1 (54 +/- 15%) in adipocytes from type 2 diabetic subjects compared to controls with a further significant reduction in expression after long-term treatment (30 +/- 12%) compared to short-term treatment. IRS-2 expression also showed a significant reduction under hyperinsulinaemic conditions (20 +/- 2%) in diabetics vs. controls. Furthermore, long-term treatment with insulin in diabetic adipocytes led to a significant reduction in the phosphorylation of IRS-1(68 +/- 11%), IRS-2 (82 +/- 11%), Akt (42 +/- 2%), and MAPK (92 +/- 12%) and in the subsequent association between p85 to IRS-1 and IRS-2 (100 +/- 16% and 96 +/- 12%) in comparison to controls. Investigating glucose uptake diabetic adipocytes revealed a significant reduction of 90 +/- 2%. In this study, we were able to establish a new diabetic model of primary human adipocytes. A defect in early insulin signalling in type 2 diabetic patients under hyperinsulinaemic conditions was determined. These results might help to give further insights in early insulin action; additionally, this human model represents a useful target for the study of new therapeutic approaches.
KW - Adipocytes
KW - Aged
KW - Case-Control Studies
KW - Cell Differentiation
KW - Cells, Cultured
KW - Deoxyglucose
KW - Diabetes Mellitus, Type 2
KW - Female
KW - Humans
KW - Hyperinsulinism
KW - Insulin
KW - Insulin Receptor Substrate Proteins
KW - Intracellular Signaling Peptides and Proteins
KW - Male
KW - Middle Aged
KW - Mitogen-Activated Protein Kinases
KW - Phosphatidylinositol 3-Kinases
KW - Phosphoproteins
KW - Phosphorylation
KW - Protein-Serine-Threonine Kinases
KW - Proto-Oncogene Proteins
KW - Proto-Oncogene Proteins c-akt
KW - Signal Transduction
KW - Time Factors
U2 - 10.1055/s-2004-825798
DO - 10.1055/s-2004-825798
M3 - SCORING: Journal article
C2 - 15326562
VL - 36
SP - 531
EP - 537
JO - HORM METAB RES
JF - HORM METAB RES
SN - 0018-5043
IS - 8
ER -