Fig. 8.

Enhanced induction of monocyte CD36 expression on coincubation with VSMCs under diabetic conditions. A and B: THP-1 cells were cocultured for 24 h with HVSMCs that were pretreated with medium alone or medium containing HG or S100B for 3 days. As a control, THP-1 cells were either left alone (basal) or treated for 24 h with HG alone (HG) or S100B alone (S100B). Cell surface expression of CD36 in both control THP-1 cells and THP-1 cells cocultured with HVSMCs was analyzed by flow cytometry. A: representative flow cytometry data showing percent changes in cell surface expression of CD36 in THP-1 cells. B: bar graph shows mean CD36 fluorescence intensity, expressed as relative fluorescence units, of THP-1 cells under the indicated conditions. *P < 0.05; **P < 0.01; ***P < 0.001 vs. basal. #P < 0.001; $P < 0.05 vs. coculture without HG or S100B. Man/Coculture, CD36 expression in THP-1 monocytes cocultured with osmotic control mannitol (5.5 mM glucose + 19.5 mM mannitol). C: mouse monocytes (WEHI78/24) were cocultured with db/+ or db/db MVSMCs for 30 min. Bound WEHI78/24 cells were collected, and CD36 mRNA expression was analyzed by real-time quantitative RT-PCR using CD36 and β-actin (internal control) primers. Bar graph shows CD36 mRNA normalized to 18S. *P < 0.0001 vs. db/+. D: bound WEHI78/24 cells were also fixed and immunostained with CD36 antibody. Fluorescence due to CD36 immunostaining was quantified and expressed as a percentage of WEHI78/24 cells bound to db/+ cells. Values are means ± SE (n = 3). *P < 0.01.