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. 2013 Jul 15;123(8):3600–3613. doi: 10.1172/JCI65592

Figure 5. Low-dose FK506 reverses monocrotaline-induced PAH in rats, inhibits proliferation, and induces apoptosis in PASMCs.

Figure 5

(A) RVSP (mmHg) in rats 21 days after monocrotaline (MCT) treatment (60 mg s.c. once), with or without 3-week treatment with vehicle (VEH) or low-dose FK506 (n = 6–8; **P < 0.01 compared with VEH treated; #P < 0.05 compared with day 21 monocrotaline; §§§P < 0.001 compared with day 42 monocrotaline vehicle, 1-way ANOVA, Bonferroni post test). (B) RVH (Fulton index) in same group as above (***P < 0.001 compared with vehicle treated; §P < 0.05 compared with day 42 monocrotaline vehicle, 1-way ANOVA, Bonferroni post test). (C) Represented lung histology (Movat stain) (scale bar: 100 μm). (D) MTT proliferation assay with PASMCs in the presence or absence of PDGF (20 ng/ml) 72 hours after stimulation with vehicle (CON), BMP4 (10 ng/ml), or FK506 (0.2 and 2.0 ng/ml) (n = 5; **P < 0.01, ***P < 0.001, BMP4 vs. CON [in the presence of PDGF], 1-way ANOVA, Dunnett’s). (E) Assessment of apoptosis with measurement of caspase-3/7 luminescence under 24-hour serum starvation or treatment with BMP4 (10 ng/ml) or FK506 (0.2, 1.0, 2.0, and 15 ng/ml) (n = 5; *P < 0.05 1-way ANOVA, Dunnett’s post test). (F) Luciferase NFAT reporter activation assay in PASMCs stimulated with PDGF (30 ng/ml) and doses of FK506 between 0.2 and 15 ng/ml. (##P < 0.01 and ###P < 0.001, vs. stimulation by PDGF alone, 1-way ANOVA with Bonferroni’s multiple pairs test). (G) Il2/Gapdh mRNA expression in whole lungs of rats 21 days after monocrotaline treatment (60 mg s.c. once), with or without 3-week treatment with vehicle or low-dose FK506 (n = 6–8; nonsignificant, 2-way ANOVA). Mean ± SEM.