Objective:

2

To verify the effect of Buyang Huanwu Decoction (BHD) in ameliorating erectile dysfunction (ED) after radical prostatectomy (RP).

Methods:

2

The composition of BHD was verified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) analysis. Bilateral cavernous nerve crush injury (BCNI) in rats was used to mimic the neurovascular injury occurring after RP. By the envelope method

forty rats were randomly divided into 4 groups as follows: sham (cavernous nerves exposed only)

model (BCNI)

low-dosage BHD [LBHD

12.8 g/(kg•d)]

and high-dosage BHD [HBHD

51.2 g/(kg•d)] groups

10 rats in each group

feeding for 3 weeks respectively. Erectile function was evaluated by measuring intracavernosal pressure (ICP). Changes in the histopathology of corpus cavernosum (CC) were examined by hematoxylin-eosin staining. Meanwhile

the fibrosis of CC was measured by Masson's trichrome staining and Western blot was used to detect the expressions of collagen Ⅰ

transforming growth factor beta 1 (TGF-β1) and α-smooth muscle actin (α-SMA). Apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and Western blot for determining the expressions of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax). The oxidative stress in the CC were assessed by the superoxide dismutase (SOD)

malondialdehyde (MDA) and reactive oxygen species (ROS) levels. The proteins expression of c-Jun N-terminal kinase (JNK) and c-Jun were detected by Western blot. In addition

the expression of α-SMA and p-c-Jun in the CC was observed by double immunofluorescence staining.

Results:

2

The UPLC-QTOF-MS/MS analysis showed that BHD contained calycosin-7-O-β-D-glucoside

ononin

calycosin and formononetin. Compared with the model group

LBHD and HBHD treatment improved the ICP and the circumference

area

and weight of CC (

P

<

0.05 or

P

<

0.01). Furthermore

LBHD and HBHD treatments increased CC smooth muscle content and decreased apoptosis index (

P

<

0.05 or

P

<

0.01). LBHD and HBHD also elevated SOD and expression level of α-SMA and Bcl-2

and reduced MDA and ROS levels

as well as expression of TGF-β1

collagen Ⅰ

Bax

p-c-JNK

p-JNK in the CC compared with the model group (

P

<

0.05 or

P

<

0.01). The double immunofluorescence staining showed that the fluorescence degree of p-c-Jun in both LBHD and HBHD treatment groups was significantly reduced

whereas the α-SMA expression increased (

P

<

0.05 or

P

<

0.01).

Conclusions:

2

BHD can improve ED of rats with BCNI

which is related to inhibiting fibrosis

apoptosis

and oxidative stress of CC. The ROS/JNK/c-Jun signaling pathway may play an important role in the process.