Fig 1.
Cavernous nerve (CN) traction model.
The CN is separated from the surface of prostate between the major pelvic ganglion (MPG) and apex of the prostate, and then pulled by a glass hook with a force of 0.2 Newton.
Fig 2.
Comparison of erectile function in each group.
(A) Representative recordings of intracavernosal pressure (ICP) and arterial pressure during 1-min electrical stimulation at 2.5V, 5.0V and 7.5V. (B,C,D) The ratio of ICP to mean arterial pressure (MAP) for each group, at 2.5V, 5.0V and 7.5V. N = 8 in each group. * P<0.05 compared with control group. # P<0.05 compared with 2-min CN crush group.
Fig 3.
Changes of neuronal nitric oxide synthase(nNOS) and neurofilament content in the corpus cavernosum and MPG.
(A) Neurofilament staining (green) and nNOS staining (red) in the dorsal penile nerve from different groups were performed with immunofluorescence. Nuclei were stained with DAPI (blue). Magnification: 400×. (B,C) Ratio of neurofilament positive area to the whole dorsal penile nerve area and nNOS positive area to the whole dorsal penile nerve area of four groups. Data are shown in the form of percentage. (D) Representative western blot bands for nNOS in the corpus cavernosum and MPG respectively. (E,F) Relative density of nNOS compared with β-actin in the corpus cavernosum and MPG. Data are shown as the fold changes over the control group. N = 6 in each group.* P<0.05 compared with control group. # P<0.05 compared with 2-min CN crush group.
Fig 4.
Changes of smooth muscle and collagen content in corpus cavernosum.
(A) Smooth muscle staining (green) in corpus cavernosum in each group using immunofluorescence with anti-α-smooth muscle actin (α-SMA) antibody. Nuclei were stained with DAPI (blue). Magnification: 200×. (B) Ratio of α-SMA positive area to the whole cavernous tissue area. Data are shown in the form of percentage. (C) Masson’s trichrome staining of corpus cavernosum: smooth muscle cells are stained red, and collagen fibers are stained blue. (D) Ratio of collagen to smooth muscle, reflecting the level of fibrosis in penile tissue. N = 6 in each group. * P<0.05 compared with control group. # P<0.05 compared with 2-min CN crush group.
Fig 5.
Changes of the transforming growth factor-β1 (TGF-β1)–Smad2/3 pathway in corpus cavernosum.
(A) The expression of TGF-β1, Smad2/3 and phosphorylated Smad2/3 (p-Smad2/3) in penile tissue detected by western blot. (B,C,D) Relative density of TGF-β1, p-Smad2/3, and Smad2/3 compared with β-actin. Data are shown as the fold changes over the control group. N = 6 in each group. * P<0.05 compared with control group. # P<0.05 compared with 2-min CN crush group.
Fig 6.
Changes of the apoptosis in corpus cavernosum.
(A) The expression of Bcl-2 and Bax detected by western blot. (B) Ratio of Bax to Bcl-2. Data are shown as the fold changes over the control group. (C) Bar graph depicts caspase3 activity in penile tissues measured by a Caspase3 Activity Assay Kit, which was calculated according to a standard curve and normalized by the protein concentration. N = 6 in each group. * P<0.05 compared with control group. # P<0.05 compared with 2-min CN crush group.