Reduction of intracellular cholesterol accumulation in THP-1 macrophages by a combination of rosiglitazone and atorvastatin.

Rosiglitazone and atorvastatin combination therapy has beneficial effects on both glycemic control and plasma lipid levels in type 2 diabetic patients. In the present study, we sought to determine whether this combination can also exert direct antiatherosclerotic effects in macrophages. Our results show that 2 microM rosiglitazone, alone or combined with 5 microM atorvastatin, significantly upregulated the expression of the ATP-binding cassette transporter ABCA1 and of the class B scavenger receptor CLA-1 (CD36 and LIMPII analog), both involved in cholesterol efflux from macrophages. On the other hand, the combination with atorvastatin attenuated the inductive response elicited by rosiglitazone alone on CD36 mRNA (34%, P < 0.05) and protein (16%, P < 0.05), while the uptake of oxidized low density lipoprotein (LDL) remained unaffected. When we examined the effects of the drugs on acetyl-LDL-induced cholesterol accumulation, we found that only the combination of atorvastatin with rosiglitazone caused a net depletion in the cholesteryl ester content of macrophages (35%, P < 0.05). Our data suggest that this reduction was not mediated by effects on proteins that regulate cholesterol flux, but it may be related to the inhibition of cholesteryl ester formation elicited by the statin.

[1]  P. Weber,et al.  Lovastatin decreases the receptor-mediated degradation of acetylated and oxidized LDLs in human blood monocytes during the early stage of differentiation into macrophages. , 1999, Arteriosclerosis, thrombosis, and vascular biology.

[2]  G. L. Mills,et al.  A guidebook to lipoprotein technique , 1984 .

[3]  M. Vázquez-Carrera,et al.  Avasimibe and atorvastatin synergistically reduce cholesteryl ester content in THP-1 macrophages. , 2002, European journal of pharmacology.

[4]  H. Kempen,et al.  Vastatins inhibit cholesterol ester accumulation in human monocyte-derived macrophages. , 1991, Arteriosclerosis and thrombosis : a journal of vascular biology.

[5]  F. Bernini,et al.  HMG-CoA reductase inhibitors reduce acetyl LDL endocytosis in mouse peritoneal macrophages. , 1995, Arteriosclerosis, thrombosis, and vascular biology.

[6]  T. Langmann,et al.  Molecular cloning of the human ATP-binding cassette transporter 1 (hABC1): evidence for sterol-dependent regulation in macrophages. , 1999, Biochemical and biophysical research communications.

[7]  E. Ros,et al.  Walnut-enriched diet increases the association of LDL from hypercholesterolemic men with human HepG2 cells. , 2001, Journal of lipid research.

[8]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[9]  J. Auwerx,et al.  Reduction of atherosclerosis in apolipoprotein E knockout mice by activation of the retinoid X receptor , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. McManus,et al.  ABCA1 mRNA and Protein Distribution Patterns Predict Multiple Different Roles and Levels of Regulation , 2002, Laboratory Investigation.

[11]  M. Jaye,et al.  PPAR-α and PPAR-γ activators induce cholesterol removal from human macrophage foam cells through stimulation of the ABCA1 pathway , 2001, Nature Medicine.

[12]  S. Perrey,et al.  Troglitazone Inhibits Atherosclerosis in Apolipoprotein E–Knockout Mice: Pleiotropic Effects on CD36 Expression and HDL , 2001, Arteriosclerosis, thrombosis, and vascular biology.

[13]  A. von Eckardstein,et al.  Differential effects of lovastatin on the trafficking of endogenous and lipoprotein-derived cholesterol in human monocyte-derived macrophages. , 1998, Arteriosclerosis, thrombosis, and vascular biology.

[14]  Yong Ji,et al.  Scavenger Receptor BI Promotes High Density Lipoprotein-mediated Cellular Cholesterol Efflux* , 1997, The Journal of Biological Chemistry.

[15]  S. Haffner,et al.  Effects of Diabetes and Level of Glycemia on All-Cause and Cardiovascular Mortality: The San Antonio Heart Study , 1998, Diabetes Care.

[16]  S. Horiuchi,et al.  Differential effects of an acyl‐coenzyme A: cholesterol acyltransferase inhibitor on HDL‐induced cholesterol efflux from rat macrophage foam cells , 1995, FEBS letters.

[17]  C. Argmann,et al.  Activation of Peroxisome Proliferator–Activated Receptor Gamma and Retinoid X Receptor Results in Net Depletion of Cellular Cholesteryl Esters in Macrophages Exposed to Oxidized Lipoproteins , 2003, Arteriosclerosis, thrombosis, and vascular biology.

[18]  D. Altshuler,et al.  The role of PPAR-γ in macrophage differentiation and cholesterol uptake , 2001, Nature Medicine.

[19]  R. Evans,et al.  PPARγ Promotes Monocyte/Macrophage Differentiation and Uptake of Oxidized LDL , 1998, Cell.

[20]  R. Evans,et al.  Oxidized LDL Regulates Macrophage Gene Expression through Ligand Activation of PPARγ , 1998, Cell.

[21]  T. Hayek,et al.  Atorvastatin therapy in hypercholesterolemic patients suppresses cellular uptake of oxidized-LDL by differentiating monocytes. , 2002, Atherosclerosis.

[22]  A. Tedgui,et al.  CLA-1/SR-BI is expressed in atherosclerotic lesion macrophages and regulated by activators of peroxisome proliferator-activated receptors. , 2000, Circulation.

[23]  M. Brown,et al.  Degradation of cationized low density lipoprotein and regulation of cholesterol metabolism in homozygous familial hypercholesterolemia fibroblasts. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Ye,et al.  Microarray analysis of peroxisome proliferator-activated receptor-gamma induced changes in gene expression in macrophages. , 2003, Biochemical and biophysical research communications.

[25]  S. Devaraj,et al.  α-Tocopherol decreases CD36 expression in human monocyte-derived macrophages , 2001 .

[26]  L. Bernier,et al.  Effect of Atorvastatin on ApoE and ApoC-I Synthesis and Secretion by THP-1 Macrophages , 2003, Journal of cardiovascular pharmacology.

[27]  J. Lehmann,et al.  An Antidiabetic Thiazolidinedione Is a High Affinity Ligand for Peroxisome Proliferator-activated Receptor γ (PPARγ) (*) , 1995, The Journal of Biological Chemistry.

[28]  K. Matsumoto,et al.  Expression of human scavenger receptor class B type I in cultured human monocyte-derived macrophages and atherosclerotic lesions. , 1999, Circulation research.

[29]  S. Walker,et al.  Quantitative RT-PCR : Pitfalls and Potential , 1999 .

[30]  R. Lorenz,et al.  TGF-beta1 downregulates CD36 and scavenger receptor A but upregulates LOX-1 in human macrophages. , 2000, American journal of physiology. Heart and circulatory physiology.

[31]  S. Grundy,et al.  Diabetes and cardiovascular disease: a statement for healthcare professionals from the American Heart Association. , 1999, Circulation.

[32]  J. Brunzell,et al.  Effects of rosiglitazone alone and in combination with atorvastatin on the metabolic abnormalities in type 2 diabetes mellitus. , 2002, The American journal of cardiology.

[33]  M. Alegret,et al.  Differential effects of peroxisome proliferator-activated receptor activators on the mRNA levels of genes involved in lipid metabolism in primary human monocyte-derived macrophages. , 2003, Metabolism: clinical and experimental.

[34]  R. Evans,et al.  PPAR-γ dependent and independent effects on macrophage-gene expression in lipid metabolism and inflammation , 2001, Nature Medicine.

[35]  C. Glass,et al.  Conditional Disruption of the Peroxisome Proliferator-Activated Receptor γ Gene in Mice Results in Lowered Expression of ABCA1, ABCG1, and apoE in Macrophages and Reduced Cholesterol Efflux , 2002, Molecular and Cellular Biology.

[36]  I. Björkhem,et al.  Elimination of Cholesterol in Macrophages and Endothelial Cells by the Sterol 27-Hydroxylase Mechanism , 1997, The Journal of Biological Chemistry.

[37]  K. Moore,et al.  Scavenger Receptors Class A-I/II and CD36 Are the Principal Receptors Responsible for the Uptake of Modified Low Density Lipoprotein Leading to Lipid Loading in Macrophages* , 2002, The Journal of Biological Chemistry.

[38]  H. Kruth,et al.  Apolipoprotein E Produced by Human Monocyte-derived Macrophages Mediates Cholesterol Efflux That Occurs in the Absence of Added Cholesterol Acceptors* , 1996, The Journal of Biological Chemistry.

[39]  A. Vaughan,et al.  The Tangier disease gene product ABC1 controls the cellular apolipoprotein-mediated lipid removal pathway. , 1999, The Journal of clinical investigation.

[40]  W. Hsueh,et al.  Troglitazone Inhibits Formation of Early Atherosclerotic Lesions in Diabetic and Nondiabetic Low Density Lipoprotein Receptor–Deficient Mice , 2001, Arteriosclerosis, thrombosis, and vascular biology.

[41]  E. Donetti,et al.  Non-lipid-related effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors. , 1996, Cardiology.

[42]  R. Evans,et al.  A PPARγ-LXR-ABCA1 Pathway in Macrophages Is Involved in Cholesterol Efflux and Atherogenesis , 2001 .

[43]  H. Sone,et al.  PPARγ ligands, troglitazone and pioglitazone, up‐regulate expression of HMG‐CoA synthase and HMG‐CoA reductase gene in THP‐1 macrophages , 2002 .

[44]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[45]  Andrew C. Li,et al.  Peroxisome proliferator–activated receptor γ ligands inhibit development of atherosclerosis in LDL receptor–deficient mice , 2000 .

[46]  G. Schmitz,et al.  ABCG1 (ABC8), the human homolog of the Drosophila white gene, is a regulator of macrophage cholesterol and phospholipid transport. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[47]  T. Rea,et al.  HMG-CoA reductase and ACAT inhibitors act synergistically to lower plasma cholesterol and limit atherosclerotic lesion development in the cholesterol-fed rabbit. , 1998, Atherosclerosis.