Figure 7: Stage-Specific Effects of ApoA-I Infusions on the Anti-Inflammatory Effects and Cholesterol Efflux Capacity of Mouse HDL Ex Vivo
2019-06-11T07:08:43Z (GMT) by
HCAECS were incubated with rHDL, apoA-I, mouse HDL (50 μg/ml), or PBS (vehicle) for 16 h, then stimulated with TNF-α (0.6 ng/ml) for 4 h. RNA was isolated to determine VCAM-1 (A) and MCP-1 (B) mRNA using RT-PCR. To measure cholesterol efflux (C), J774A.1 mouse macrophages were labeled with 8 μCi/ml [14C]cholesterol for 24 h, then treated with rHDL, apoA-I, mouse HDL (50 μg/ml), or PBS (vehicle) for 16 h. Radioactivity was measured in the supernatant and cell lysates using a liquid scintillation counter. Cholesterol efflux was determined as a percentage of secreted versus intracellular [14C]cholesterol. Data are mean ± SEM; n = 6 mice HDL/treatment group performed in duplicate. *p ≤ 0.05 compared with TNF-α–only controls, #p ≤ 0.05 compared with cells treated with mouse HDL from saline-infused mice, γp ≤ 0.05 compared with cells treated with mouse HDL from apoA-I–infused early-stage mice. HCAEC = human coronary artery endothelial cell; HDL = high-density lipoprotein; PBS = phosphate-buffered saline; rHDL = reconstituted high-density lipoprotein; RT-PCR = reverse-transcription polymerase chain reaction; TNF = tumor necrosis factor; other abbreviations as in Figure 1.