Figure 6: Flunarizine lowers Angiopoietin-2 (Angpt-2) and vascular inflammation in vivo.

Adult male C57BL/6 J mice were pretreated daily with Flunarizine (25 mg/kg body weight (bw) p.o.) or vehicle for three days before the injection with 17.5 mg/kg bw gram-negative endotoxin (lipopolysaccharides [LPS] from Escherichia coli i.p.) or control. Expression of pulmonary mRNA and Angpt-2 serum levels were measured 12 h after LPS (n = 3–9). (A) Real-time polymerase chain reaction (RT-PCR) from lung homogenates for tumor necrosis factor α (TNFα). (B) RT-PCR from lung homogenates for Interleukin-6 (IL-6). (C) RT-PCR from lung homogenates for Angpt-2. (D) Angpt-2 serum levels were quantified by enzyme-linked immunosorbent assay (ELISA). (E) RT-PCR from lung homogenates for intercellular adhesion molecule-1 (ICAM-1). (F) Immunohistochemistry from murine lungs for Gr-1 (red), endothelial-specific lectin (green) and nuclear staining (4′,6-diamidino-2-phenylindole, blue) (G) Kaplan Meier Survival after LPS administration (20 mg/kg bw) in mice that were pretreated with 25 mg/kg bw FLU or vehicle for three days once a day (n = 8–10, p = 0.408). Columns are presented as mean ± SEM.