- Email: [email protected]
W08-P-024 A human adipocyte cell system as a tool to study cellular lipid metabolism
Workshops W8 Visceral fat, adipocytokines and cardiovascular risk
IW08-P-023]
W08-P-021 / THE IMPACT OF ADIPOCYTOKINES AND FAT COMPARTMENTS ON DYSLIPIDAEMIA AND INSULIN RESISTANCE: AN INDEPENDENT ROLE FOR ADIPONECTIN? T. Ng 1, M. Farvid 2, D. Chan 1, E Barrett 1, G. Watts 1. 1School of Medicine
and Pharmacology, University of Western Australia, Perth, Australia; 2Shaheed Beheshti University of Medical Sciences, Tehrarg Iran Objective: To investigate the relationship of plasma adipocytokines and body fat compartments with insulin sensitivity and plasma lipids in 41 men with wide ranging BMI (22 - 35 kg/m2). Methods: Plasma adipocytokine levels were measured using enzyme immunoassays. Insulin resistance was estimated by homeostasis model assessment (HOMA) score. Visceral and subcutaneous abdominal adipose tissue masses were determined using MRI and total adipose tissue mass (TATM) with bioelectrical impedance. Results: In univafiate regression, plasma adipouectin, leptin, visceral, subcutaneous, and total adipose tissue masses were associated with plasma triglyceride, HDL-cholesterol levels, and the HOMA score. In multivariate regression, plasma adipouectiu and visceral fat were independent predictors for plasma triglyceride levels and the HOMA score. LDL-cholesterol levels were not significantly associated with adipose tissue mass or plasma adipocytokine levels. Plasma resistin, TNF-~ and IL-6 levels were not significantly associated with plasma lipids or the HOMA score. Conclusion: In men, dyslipidaemia and insulin resistance appear to be determined by both visceral fat and plasma adiponectin levels. ]
W08-P-022 / SERUM ADIPONECTIN - ITS ASSOCIATION W I T H INFLAMMATION AND STATIN TREATMENT A.G. Olsson, E Cherfan, H. Blomqvist, L. Jonassou. Dpt of Medicine and
Care, Faculty of Health Sciences, Linkb'ping University, Linkb'ping, Sweden Objective: Adiponectin, a recently discovered adipocytokine, has potential anti-atherogenic and anti-inflammatory effects, as shown in experimental studies. An inverse relationship between adipouectin and C-reactive protein (CRP) has been demonstrated in clinical studies. In the present study, we measured serum levels of adiponnctin in non-diabetics with stable coronary artery disease (CAD) and related the findings to inflammatory and metabolic status. We also investigated the effects of simvastatin on serum adiponectin in a randomized study. Methods: In the cross-sectional study, 50 patients with stable angina and angiographically verified CAD and 50 healthy controls were included. In the randomized, double-blind and placebo-controlled study, 80 healthy male volonteers with light to moderate hyporcholesterolemia were assigned to receive either simvastatin 40 mg or placebo for 6 weeks. Adiponectiu, CRP and interleukin-6 (IL-6) in serum were determined by ELISA. Results: Ninety-six % of the CAD patients were on long-term treatment with simvastatin. They had significantly lower levels of adiponectin, LDL, HDL and CRP compared with controls (see Table). Adiponectin was related to HDL but not to CRP or IL-6. In the randomized study, simvastatin treatment led to significant decreases in LDL and CRP compared with placebo treatment (- 43% and - 21%, respectively). The simvastatiu group had a 16% reduction in adiponectin whereas the placebo group had a 14% increase. The difference in adiponectiu between groups was significant (p =
o.o4). Adiponcctin mg/l LDL mmol/l HDL mmol/l CRP mg/l IL-6 pg/l
Pat (n = 50)
Contr (n = 50)
p<
6.6 (4.8-9.7) 2.6 (2.2-3.0) 1.2 (1.0-1.4) 1.0 (0.6-2.1) 1.8 (1.5-3.2)
8.8 (5.5-14.6) 4.0 (3.5-4.5) 1.5 (1.2-1.7) 2.0 (1.0-3.3) 1.8 (1.4-2.9)
0.01 0.001 0.01 0.05 NS
Values are given as median (interquartile range). Conclusion: In CAD patients with 96% simvastatin treatment, serum adiponectin was low in spite of low CRP levels. In a randomized study, simvastatin reduced CRP but tended to decrease adiponectin. The results indicate the need of complementary treatment, consisting of intensified life style modifications and/or addition of new drags, in CAD patients.
35
ADIPONECTIN AND INFLAMMATORY MARKERS IN PATIENTS W I T H CORONARY ARTERY DISEASE
G. Olsson, H. Blomqvist, L. Jonasson. Internal medicine, Dept of Medicine and Care, Health Sciences, Linl~ping, Sweden Objectives: Low levels of the adipocyte-derived hormone adiponectin have been associated with diabetes, metabolic syndrome, inflammation and atherogenesis. We measured adiponectin in patients with stable coronary artery disease (CAD) and related the findings to inflammatory status in order to better understand the possible mechanism behind the antiatherogenic properties of adiponectin. Methods: Fifty non-diabetic patients with angiographically verified CAD, free of unstable symptoms the last 6 months, were included as well as 50 clinically healthy controls. Forty-nine patients had received more than 6 months treatment with statins. Serum levels of adiponectin and inflammatory markers were quantified by enzyme-linked immunosorbent assay (ELISA). Results: Patients and controls did not differ regarding age, sex, blood pressure, smoking status, fasting glucose, HDL cholesterol. Patients had lower mean LDL cholesterol and triglyceride levels than controls. Mean adiponectin levels were significantly decreased in the patient group compared to controls (7.5 +0.5 (SEM) ttg/ml vs. 10.1 -t-0.8 ttg/ml, p=0.01). Adiponectin showed a significant negative correlation to the chemokine GRO-c~, which is thought to promote monocyte arrest on the endothelium. Adiponectin levels were not related to any of the other laboratory or clinical parameters. Inflammatory markers hsCRP, VCAM-1, ICAM-1, P-selectin, MCP-1 and GRO-alpha did not differ between patients and controls. Conclusion: Stable CAD patients had significantly decreased levels of adiponectin although they did not show any signs of systemic inflammation or impaired metabolic status. The data supports the idea of low adiponectin as a strong indicator of atherosclerosis even in the absence of inflammation. HUMAN ADIPOCYTE CELL SYSTEM AS A IWOO-P-O24JATOOL TO STUDY CELLULAR LIPID METABOLISM J. Prawitt 1, J. Heeren 1, A. Niemeier 2, M. Kassem 3, U. Beisiegel 1. I1BM
li: lnatitute for Molecular Cell Biology, University Hospital Hamburg- Epp endo rf, Germany, 2Department of Orthopaedics, University Hospital Hamburg-Eppendo~ Germany, 3Department of Endocrinology, University Hospital Odenae, Denmark Objective: The increase of adipose tissue is connected with the manifestation of a wide range of diseases (e.g. diabetes, the metabolic syndrome). Since there are only very few and insufficient cell models available to study the metabolism of the human adipocyte under physiological and pathological conditions, we have established a human adipocyte cell culture system which allows us to analyze the lipid and lipoprotein metabolism with regard to insulin responsiveness in a human system. Methods: Cells of the human mesenchymal stem cell line hMSC-Tert (human marrow stromal cells - telomerase immortalized) are differentiated into mature adipocytes over a period of 21 days. The differentiated state is confirmed by morphological means (oil red oil staining), by RT-PCR detection of adipocyte specific markers and by analysis of intracellular lipid parameters. Results: hMSC-Tert cells were isolated from human bone marrow aspirates and immortalized by insertion of human telomerase. These stem cells have been shown to exhibit a normal karyotype [1]. We have successfully established a differentiation protocol which produces mature adipocytes from hMSC-Tert cells. After differentiation, these cells show advanced accumulation of lipid droplets and express a typical pattern of adipocyte markers (e.g. aP2, LPL, PPARv, apoE). Intracellular triglyceride levels are increased. In ongoing studies we will analyze the insulin sensitivity of the cell system, furthermore its lipid and lipoprotein metabolism with respect to insulin signalling. Conclusion: We have established a human adipose cell system which appears to be a well-functioning model of the human adipocyte. This cell model will be valuable to study the lipid and lipoprotein metabolism of human adipose tissue under set conditions and to elucidate the underlying molecular mechanisms.
Literature: [I] J. Simonsen, C. Rosada, N. Serakinci, J. Justesen, K. Stenderup, S. Rattan, T. Jensen, M. Kassem: Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells.Nature Biotechnology 2002 20:592-6
75th EAS Congress, 23-26 April 2005, Prague, Czech Republic
0 I 0 Go
IW08-P-023]
W08-P-021 / THE IMPACT OF ADIPOCYTOKINES AND FAT COMPARTMENTS ON DYSLIPIDAEMIA AND INSULIN RESISTANCE: AN INDEPENDENT ROLE FOR ADIPONECTIN? T. Ng 1, M. Farvid 2, D. Chan 1, E Barrett 1, G. Watts 1. 1School of Medicine
and Pharmacology, University of Western Australia, Perth, Australia; 2Shaheed Beheshti University of Medical Sciences, Tehrarg Iran Objective: To investigate the relationship of plasma adipocytokines and body fat compartments with insulin sensitivity and plasma lipids in 41 men with wide ranging BMI (22 - 35 kg/m2). Methods: Plasma adipocytokine levels were measured using enzyme immunoassays. Insulin resistance was estimated by homeostasis model assessment (HOMA) score. Visceral and subcutaneous abdominal adipose tissue masses were determined using MRI and total adipose tissue mass (TATM) with bioelectrical impedance. Results: In univafiate regression, plasma adipouectin, leptin, visceral, subcutaneous, and total adipose tissue masses were associated with plasma triglyceride, HDL-cholesterol levels, and the HOMA score. In multivariate regression, plasma adipouectiu and visceral fat were independent predictors for plasma triglyceride levels and the HOMA score. LDL-cholesterol levels were not significantly associated with adipose tissue mass or plasma adipocytokine levels. Plasma resistin, TNF-~ and IL-6 levels were not significantly associated with plasma lipids or the HOMA score. Conclusion: In men, dyslipidaemia and insulin resistance appear to be determined by both visceral fat and plasma adiponectin levels. ]
W08-P-022 / SERUM ADIPONECTIN - ITS ASSOCIATION W I T H INFLAMMATION AND STATIN TREATMENT A.G. Olsson, E Cherfan, H. Blomqvist, L. Jonassou. Dpt of Medicine and
Care, Faculty of Health Sciences, Linkb'ping University, Linkb'ping, Sweden Objective: Adiponectin, a recently discovered adipocytokine, has potential anti-atherogenic and anti-inflammatory effects, as shown in experimental studies. An inverse relationship between adipouectin and C-reactive protein (CRP) has been demonstrated in clinical studies. In the present study, we measured serum levels of adiponnctin in non-diabetics with stable coronary artery disease (CAD) and related the findings to inflammatory and metabolic status. We also investigated the effects of simvastatin on serum adiponectin in a randomized study. Methods: In the cross-sectional study, 50 patients with stable angina and angiographically verified CAD and 50 healthy controls were included. In the randomized, double-blind and placebo-controlled study, 80 healthy male volonteers with light to moderate hyporcholesterolemia were assigned to receive either simvastatin 40 mg or placebo for 6 weeks. Adiponectiu, CRP and interleukin-6 (IL-6) in serum were determined by ELISA. Results: Ninety-six % of the CAD patients were on long-term treatment with simvastatin. They had significantly lower levels of adiponectin, LDL, HDL and CRP compared with controls (see Table). Adiponectin was related to HDL but not to CRP or IL-6. In the randomized study, simvastatin treatment led to significant decreases in LDL and CRP compared with placebo treatment (- 43% and - 21%, respectively). The simvastatiu group had a 16% reduction in adiponectin whereas the placebo group had a 14% increase. The difference in adiponectiu between groups was significant (p =
o.o4). Adiponcctin mg/l LDL mmol/l HDL mmol/l CRP mg/l IL-6 pg/l
Pat (n = 50)
Contr (n = 50)
p<
6.6 (4.8-9.7) 2.6 (2.2-3.0) 1.2 (1.0-1.4) 1.0 (0.6-2.1) 1.8 (1.5-3.2)
8.8 (5.5-14.6) 4.0 (3.5-4.5) 1.5 (1.2-1.7) 2.0 (1.0-3.3) 1.8 (1.4-2.9)
0.01 0.001 0.01 0.05 NS
Values are given as median (interquartile range). Conclusion: In CAD patients with 96% simvastatin treatment, serum adiponectin was low in spite of low CRP levels. In a randomized study, simvastatin reduced CRP but tended to decrease adiponectin. The results indicate the need of complementary treatment, consisting of intensified life style modifications and/or addition of new drags, in CAD patients.
35
ADIPONECTIN AND INFLAMMATORY MARKERS IN PATIENTS W I T H CORONARY ARTERY DISEASE
G. Olsson, H. Blomqvist, L. Jonasson. Internal medicine, Dept of Medicine and Care, Health Sciences, Linl~ping, Sweden Objectives: Low levels of the adipocyte-derived hormone adiponectin have been associated with diabetes, metabolic syndrome, inflammation and atherogenesis. We measured adiponectin in patients with stable coronary artery disease (CAD) and related the findings to inflammatory status in order to better understand the possible mechanism behind the antiatherogenic properties of adiponectin. Methods: Fifty non-diabetic patients with angiographically verified CAD, free of unstable symptoms the last 6 months, were included as well as 50 clinically healthy controls. Forty-nine patients had received more than 6 months treatment with statins. Serum levels of adiponectin and inflammatory markers were quantified by enzyme-linked immunosorbent assay (ELISA). Results: Patients and controls did not differ regarding age, sex, blood pressure, smoking status, fasting glucose, HDL cholesterol. Patients had lower mean LDL cholesterol and triglyceride levels than controls. Mean adiponectin levels were significantly decreased in the patient group compared to controls (7.5 +0.5 (SEM) ttg/ml vs. 10.1 -t-0.8 ttg/ml, p=0.01). Adiponectin showed a significant negative correlation to the chemokine GRO-c~, which is thought to promote monocyte arrest on the endothelium. Adiponectin levels were not related to any of the other laboratory or clinical parameters. Inflammatory markers hsCRP, VCAM-1, ICAM-1, P-selectin, MCP-1 and GRO-alpha did not differ between patients and controls. Conclusion: Stable CAD patients had significantly decreased levels of adiponectin although they did not show any signs of systemic inflammation or impaired metabolic status. The data supports the idea of low adiponectin as a strong indicator of atherosclerosis even in the absence of inflammation. HUMAN ADIPOCYTE CELL SYSTEM AS A IWOO-P-O24JATOOL TO STUDY CELLULAR LIPID METABOLISM J. Prawitt 1, J. Heeren 1, A. Niemeier 2, M. Kassem 3, U. Beisiegel 1. I1BM
li: lnatitute for Molecular Cell Biology, University Hospital Hamburg- Epp endo rf, Germany, 2Department of Orthopaedics, University Hospital Hamburg-Eppendo~ Germany, 3Department of Endocrinology, University Hospital Odenae, Denmark Objective: The increase of adipose tissue is connected with the manifestation of a wide range of diseases (e.g. diabetes, the metabolic syndrome). Since there are only very few and insufficient cell models available to study the metabolism of the human adipocyte under physiological and pathological conditions, we have established a human adipocyte cell culture system which allows us to analyze the lipid and lipoprotein metabolism with regard to insulin responsiveness in a human system. Methods: Cells of the human mesenchymal stem cell line hMSC-Tert (human marrow stromal cells - telomerase immortalized) are differentiated into mature adipocytes over a period of 21 days. The differentiated state is confirmed by morphological means (oil red oil staining), by RT-PCR detection of adipocyte specific markers and by analysis of intracellular lipid parameters. Results: hMSC-Tert cells were isolated from human bone marrow aspirates and immortalized by insertion of human telomerase. These stem cells have been shown to exhibit a normal karyotype [1]. We have successfully established a differentiation protocol which produces mature adipocytes from hMSC-Tert cells. After differentiation, these cells show advanced accumulation of lipid droplets and express a typical pattern of adipocyte markers (e.g. aP2, LPL, PPARv, apoE). Intracellular triglyceride levels are increased. In ongoing studies we will analyze the insulin sensitivity of the cell system, furthermore its lipid and lipoprotein metabolism with respect to insulin signalling. Conclusion: We have established a human adipose cell system which appears to be a well-functioning model of the human adipocyte. This cell model will be valuable to study the lipid and lipoprotein metabolism of human adipose tissue under set conditions and to elucidate the underlying molecular mechanisms.
Literature: [I] J. Simonsen, C. Rosada, N. Serakinci, J. Justesen, K. Stenderup, S. Rattan, T. Jensen, M. Kassem: Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells.Nature Biotechnology 2002 20:592-6
75th EAS Congress, 23-26 April 2005, Prague, Czech Republic
0 I 0 Go