Dihydrotestosterone modulates endothelial progenitor cell function via RhoA/ROCK pathway.
BACKGROUND
Previous findings indicate that testosterone level is negatively correlated with the incidence and mortality of cardiovascular diseases in men. Endothelial progenitor cells (EPCs) play a critical role in endothelial healing and vascular integrity. This study aimed to examine the effects of dihydrotestosterone (DHT), an active metabolite of testosterone, on human EPC function and investigate the underlying mechanism.


METHODS
EPCs were isolated from peripheral blood of healthy adult males and incubated with a series of concentrations (1, 10, and 100 nmol/L in dimethyl sulfoxide) of DHT for 24 h or with 10 nmol/L DHT for different periods (6, 12, 24, 36, and 48 h). EPC proliferation, migration, and adhesion were determined by MTT assay, modified Boyden chamber assay, and cell counting, respectively. Furthermore, vascular endothelial growth factor (VEGF) production was examined by ELISA, RhoA activity was determined through pull-down assay. The protein level of RhoA was quantified by Western blot analysis.


RESULTS
DHT significantly increased the proliferative, migratory, and adhesive abilities of EPCs in a dose- and time-dependent manner and upregulated the levels of VEGF and activated RhoA. However, RhoA inhibitor C3 exoenzyme or ROCK inhibitor Y-27632 significantly inhibited DHT-induced proliferation, migration, and adhesion, as well as VEGF production. Moreover, C3 exoenzyme inhibited the activation of RhoA stimulated by DHT.


CONCLUSIONS
DHT promotes EPC proliferation, migration, and adhesion activities via RhoA/ROCK pathway.
Emergency Medicine Center, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Hao
H
Zhang
Department of Nuclear Medicine, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Liang
L
Shi
Emergency Medicine Center, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Guo-Qing
GQ
Ren
Emergency Medicine Center, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Wen-Wen
WW
Sun
Department of Cardiology, Chunan Branch of Zhejiang Provincial People's Hospital 311700, Zhejiang, China.
Yi-Bin
YB
Wang
Emergency Medicine Center, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Yi-Kun
YK
Chen
Emergency Medicine Center, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Jiang-Ning
JN
Yin
Department of ICU, Affiliated Hospital of Jiangsu University Zhenjiang 212001, China.
Bing
B
Wan
Endothelial progenitor cells; RhoA/ROCK pathway; cardiovascular disease; dihydrotestosterone