Fig 1.
Fasudil has no effect on endothelial morphology of aorta.
Representative microscopic photographs of aorta sections stained with H&E stain (scale bar = 30 μm; n = 6). The magnified image in the right.
Table 1.
Effects of fasudil on blood biochemistry and body weight, systolic blood pressures, heart rate in normal rats.
Fig 2.
Histopathological changes in aortas when subjected to CIH.
The aorta samples from different groups were analysed histochemically. Representative aorta histology in the Normoxia, CIH, and CIH + Fa groups were shown (magnification, 400 ×).
Fig 3.
Vasodilator responses in endothelium-intact and endothelium-denuded aortas.
(A) Results were expressed as an isometric tension in endothelium-intact rat aortas. (B) Results were expressed as an isometric tension endothelium-denuded in rat aortas. (C) Relaxation responses to ACh expressed as a percentage of PE-induced pre-contraction. (D) Relaxation responses to SNP expressed as a percentage of PE-induced pre-contraction. Values were the mean ± SE. * p < 0.05, CIH group vs Normoxia group; # p < 0.05, CIH + Fa group vs CIH group (n = 6 for each group).
Fig 4.
Levels NO in the serum and aorta.
(A) NO content was measured in serum from the Normoxia, CIH and CIH + Fa groups by the Griess assay. (B) NO production was measured in aortas isolated from the Normoxia, CIH and CIH + Fa groups with a Griess assay. (C) eNOS protein was measured in aortas isolated from the Normoxia, CIH and CIH + Fa groups by Western blotting. (D) p-eNOS (Ser1177) protein was measured in aortas isolated from the Normoxia, CIH and CIH + Fa groups by Western blotting. The results were expressed as the mean ± SE. * p < 0.05, CIH group vs Normoxia group; # p < 0.05, CIH + Fa group vs CIH group (n = 6 for each group).
Fig 5.
Levels ET-1 in the serum and aorta when subjected to CIH.
(A) ET-1 content was measured in serum from the Normoxia, CIH and CIH+Fa groups by radioimmunoassay. (B) ET-1 protein levels were measured in aortas isolated from the Normoxia, CIH and CIH + Fa groups by western blot. The results were expressed as the mean ± SE. *p < 0.05, CIH group vs Normoxia group; # p < 0.05, CIH + Fa group vs CIH group (n = 6 for each group).
Fig 6.
Expression of RhoA, ROCK-2, p-MYPT1, and t-MYPT1 proteins in aortas when subjected to CIH.
(A-B) RhoA and ROCK-2 protein levels were measured in aortas from Normoxia, CIH and CIH + Fa groups by Western blot. (C) p-MYPT1 (Thr853) and t-MYPT1 protein levels were measured in aortas isolated from the Normoxia, CIH and CIH + Fa groups by Western blotting. The results were expressed as the mean ± SE. * p < 0.05, CIH group vs Normoxia group; # p < 0.05, CIH + Fa group vs CIH group (n = 6 for each group).
Fig 7.
Expression of NFATc3 protein in aortas when subjected to CIH.
NFATc3 protein levels were measured in aortas from Normoxia, CIH and CIH + Fa groups by Western blotting. The results were expressed as the mean ± SE. * p < 0.05, CIH group vs Normoxia group; # p < 0.05, CIH + Fa group vs CIH group (n = 6 for each group).
Fig 8.
Schematic illustration of RhoA/ROCK/NFATc3 pathway contributing to endothelial dysfunction induced by CIH.
CIH increased RhoA, ROCK and NFATc3 protein expression in aortas, and decreased p-eNOS protein expression, which lead to endothelial dysfunction. Fasudil could improve these changes.