Figure 5.

Effect of LXRα agonist on pristane-induced lung hemorrhage. (A) In vitro treatment of RAW-264.7 cells with GW3965 (GW, 1 µM), T0901317 (1 µM), or DMSO for 24 h. Oxygen consumption rate (OCR) was measured (XF96 Analyzer). Experimental treatments were performed with six technical replicates. **P < 0.01; ***P < 0.001 vs. control (Student’s unpaired t-test). (B) Adherent peritoneal Mϕ from pristane-treated B6 mice were incubated for 24-h with GW3965, T0901317, or DMSO followed by measurement of OCR. Experimental treatments were performed with six technical replicates. **P < 0.01 vs. control (Student’s unpaired t-test). (C–E), B6 mice were injected once with pristane and treated i.p. with T0901317 (200 μg/mouse/day) or DMSO (n = 10) starting on the day of pristane treatment. One group received T0901317 daily from d1–d14 (n = 10), another from d1–d3 (n = 15), another from d3–d14 (n = 11), and another from d7–d14 (n = 6). (C) Frequency of lung hemorrhage in the four groups. 5/10 control mice and 0/10 mice treated with T0901317 (d1–d14) developed DAH (**P < 0.01, χ²). (D, E) Flow cytometry of CD11b⁺CD138⁺ Mϕ from mice treated with pristane plus T0901317 (d1–d14) vs. DMSO (Control). MFI, mean fluorescence intensity. (D) Intracellular staining for Abca1 in CD11b⁺CD138⁺ cells. **P < 0.01 vs. control (Welch’s unpaired t-test). (E), Surface staining for CD11b and intracellular staining for TNFα. CD11b⁺CD138⁺ cells were gated to analyze the expression level (MFI) of CD11b and TNFα. *P < 0.05; ***P < 0.001 vs. control (Student’s unpaired t-test).