n-caproylsphingosine and Carotid-Artery-Diseases

Apoptotic and inflammatory processes occur in human arteriosclerotic lesions. We examined the hypothesis whether both processes are possibly associated by studying the colocalization of corresponding markers. In 11 human arteriosclerotic carotid arteries, proapoptotic markers (CPP32 (caspase-3), poly(ADP-ribose) polymerase, apoptosis-inducing factor, c-Jun/AP-1, and p53) and proinflammatory markers (macrophage migration inhibitory factor (MIF) and cyclooxygenase-2) were found in macrophages (MPhi) evaluated by computer-assisted immunohistomorphometry. Double-labeling studies demonstrated a colocalization of, both, proapoptotic and proinflammatory markers in these MPhi. Moreover, these MPhi also contained oxidized low-density lipoproteins (oxLDL). Exposure of cultured human MPhi to oxLDL, C6-ceramide, and tumor necrosis factor-alpha or H2O2 resulted in a significant increase of the apoptosis rate as well as of the MIF protein expression. Our study of MPhi in arteriosclerotic carotid arteries and in vitro experiments provide evidence that markers of apoptosis and inflammation are not only significantly increased but are also coexpressed. We conclude there are reciprocal modulatory interactions between apoptotic and inflammatory pathways in human plaque MPhi, which might importantly modify plaque progression or stability.

Topics: Apoptosis; Arteriosclerosis; Carotid Artery Diseases; Caspase 3; Caspases; Cells, Cultured; Ceramides; Cyclooxygenase 2; Humans; Hydrogen Peroxide; Immunohistochemistry; Lipoproteins, LDL; Macrophage Migration-Inhibitory Factors; Macrophages; Tumor Necrosis Factor-alpha

Growth differentiation factor-15/macrophage inhibitory cytokine-1 (GDF-15/MIC-1) is a new member of the transforming growth factor beta (TGF-beta) superfamily, which has most recently been found in activated macrophages (MPhi). We have now investigated GDF-15/MIC-1 in human MPhi after exposure to oxidized low-density lipoproteins (oxLDL) related mediators in vitro and in arteriosclerotic carotid arteries. Using RT-PCR and Western blotting a pronounced induction of GDF-15/MIC-1 expression by oxLDL, C6-ceramide, tumor necrosis factor (TNFalpha) and hydrogen peroxide (H2O2) was found in cultured human MPhi. In 11 human arteriosclerotic carotid arteries, immunohistochemical analyses supported by computer-assisted morphometry and regression analyses demonstrated a significant colocalization of GDF-15/MIC-1 immunoreactivity (IR) with oxLDL IR and manganese superoxide dismutase (MnSOD) IR in CD68 immunoreactive (ir) MPhi, which were also expressing AIF-IR (apoptosis-inducing factor), caspase-3-IR (CPP32), PARP-IR, c-Jun/AP-1-IR and p53-IR. Our data suggest that GDF-15/MIC-1 is inducible in human MPhi by oxLDL and its mediators in vitro and is supposed to contribute to oxidative stress dependent consequences in arteriosclerotic plaques, e.g. modulating apoptosis and inflammatory processes in activated MPhi.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Apoptosis; Apoptosis Inducing Factor; Arteriosclerosis; Bone Morphogenetic Proteins; Carotid Arteries; Carotid Artery Diseases; Caspase 3; Caspases; Cells, Cultured; Ceramides; Collagen Type XI; Flavoproteins; Growth Differentiation Factor 15; Humans; Hydrogen Peroxide; Image Processing, Computer-Assisted; Lipoproteins, LDL; Macrophages; Membrane Proteins; Oxidative Stress; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases; Proto-Oncogene Proteins c-jun; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Superoxide Dismutase; Tumor Necrosis Factor-alpha