Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease
Article Title: DBZ (Danshensu Bingpian Zhi), a Novel Natural Compound Derivative, Attenuates Atherosclerosis in Apolipoprotein E–Deficient Mice
Figure Lengend Snippet: DBZ attenuates ox‐ LDL –induced foam cell formation and promotes cholesterol efflux in macrophages. A, THP ‐1 macrophages were cotreated with the indicated concentrations of DBZ (5, 10, and 20 μmol/L) and Ox LDL (50 μg/mL) for 48 h. Lipid accumulation was observed by Oil Red O staining (scale=20 μm). B, Intracellular cholesteryl ester levels were normalized to cellular protein content in THP ‐1 macrophages. Macrophages were treated with DBZ and NBD cholesterol. Cholesterol efflux to apoA‐1 (C) and HDL (D) from THP ‐1 macrophages. Cholesterol efflux was expressed as the percentage fluorescence in the medium relative to total fluorescence. E, THP ‐1 macrophages were treated with the indicated concentrations of DBZ for 24 h. The relative protein levels of ABCA 1 and ABCG 1 were determined by Western blot analysis. T1317 (1 μmol/L) was used as a positive control. F, The transcriptional activity of LXR α was assessed by the transactivation reporter assay in 293T cells. G and H, The model structure of the complex formed by the LXR α ligand‐binding pocket and DBZ by molecular docking. I, THP ‐1 macrophages were pretreated with GGPP (10 μmol/L) for 2 h and then treated with DBZ (20 μmol/L) for 24 h. The relative protein level of ABCA 1 was determined by Western blot analysis. Prior to treatment with DBZ (20 μmol/L), THP ‐1 macrophages were treated with GGPP for 2 h and then incubated with 50 μg/mL Ox LDL for 48 h. J, Representative Oil Red O staining of THP ‐1 macrophage foam cells (scale=20 μm). K, Intracellular cholesteryl ester levels that were normalized to cellular protein content in THP ‐1 macrophages. Values are presented as the mean± SEM of at least 3 experiments ( ### P
Article Snippet: NBD cholesterol (Cayman), apoA1, high‐density lipoprotein (HDL) (ProSpec), atorvastatin (Pfizer Ltd.), lipopolysaccharide, PMA (phorbol 12‐myristate 13‐acetate), T0901317 (hereafter referred to as T1317), and GGPP (geranylgeranyl pyrophosphate) were purchased from Sigma‐Aldrich.
Techniques: Staining, Fluorescence, Western Blot, Positive Control, Activity Assay, Reporter Assay, Ligand Binding Assay, Incubation