Rho/ROCK signaling pathway and anti-cryodamage ability of human sperm.
- Author:
Jia TIAN
1
;
Shao-Hua ZHANG
2
;
Ke MA
2
;
Bao-Yun ZHAO
2
;
Bei YAN
1
;
Cheng-Bin PEI
1
;
Yue ZHOU
1
;
Hong-Hong WANG
1
;
Hong-Yan WANG
1
;
Liang-Hong MA
1
;
Xin-Zong ZHANG
3
Author Information
1. Ningxia Human Sperm Bank / Key Laboratory for Fertility Preservation and Maintenance of the Ministry of Education, General Hospital of Ningxia Medical University, Yinchuan, Ningxia 750004, China.
2. School of Clinical Medicine, Ningxia Medical University, Yinchuan, Ningxia 750004, China.
3. Guangdong Human Sperm Bank, Family Planning Hospital of Guangdong Province, Guangzhou, Guangdong 510600, China.
- Publication Type:Journal Article
- Keywords:
Rho/ROCK signaling pathway;
cryopreservation;
human sperm;
inhibitor
- From:
National Journal of Andrology
2019;25(5):322-328
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the influence of the Rho/ROCK signaling pathway on the anti-cryodamage ability of human sperm and provide some theoretical evidence for the development of high-efficiency semen cryoprotectants.
METHODS:We collected semen samples from 25 healthy males, each divided into a fresh, a normal cryopreservation control and an Rho-inhibition group. Before and after freezing, we detected sperm motility, viability, membrane integrity, morphology, DNA fragmentation index (DFI), acrosomal enzyme activity (AEA) and mitochondrial membrane potential (MMP) and determined the expressions of RhoA and ROCK proteins in the sperm by immunofluorescence staining.
RESULTS:Compared with the normal cryopreservation control, the frozen-thawed sperm of the Rho-inhibition group showed significantly increased sperm motility （ ［51.20 ± 7.70］% vs ［57.50 ± 6.83］%, P = 0.002), survival rate ( ［52.87 ± 5.07］% vs ［60.24 ± 5.53］%, P = 0.001), membrane integrity (［59.78±5.56］% vs ［67.10 ± 4.43］%, P = 0.001), percentage of morphologically normal sperm (［4.83 ± 1.11］% vs ［7.46 ± 1.28］, P = 0.001) and MMP (56.30 ± 4.28 vs 63.11 ± 2.97, P = 0.001), but decreased DFI (［27.64 ± 6.64］% vs ［18.87 ± 4.07］%, P = 0.001). There was no statistically significant difference in the AEA of the frozen-thawed sperm between the control and Rho-inhibition groups (97.65 ± 9.31 vs 98.30 ± 11.33, P > 0.05). Immunofluorescence staining revealed extensive expressions of RhoA and ROCK proteins in the head and neck of the sperm.
CONCLUSIONS:The Rho/ROCK signaling pathway plays a role in the cryodamage to human sperm, and inhibiting the activity of Rho/ROCK can significantly improve the ability of sperm to resist cryodamage.
