Expression changes of superoxide dismutase 2 and peroxiredoxin 6 in asthenospermia and their correlation with sperm motility

Zihan YANG; Xiaoling HAO; Hong WANG; Liu YE; Run HUANG; Jiangping MENG

doi:10.16016/j.1000-5404.202104129

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Expression changes of superoxide dismutase 2 and peroxiredoxin 6 in asthenospermia and their correlation with sperm motility

Basic Medicine | 更新时间：2026-01-09

- Expression changes of superoxide dismutase 2 and peroxiredoxin 6 in asthenospermia and their correlation with sperm motility
- Journal of Third Military Medical University Vol. 43, Issue 20, Pages: 2199-2207(2021)
- 作者机构：
  
  1.400016 重庆，重庆医科大学附属第一医院生殖医学中心
  2.400016 重庆，重庆医科大学检验医学院临床检验诊断学教育部重点实验室
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202104129
  CLC： R321.1;R345;R698.202
- Received：20 April 2021，
  
  Revised：2021-05-22，
  
  Published：30 October 2021
- 稿件说明：
移动端阅览
Zihan YANG, Xiaoling HAO, Hong WANG, et al. Expression changes of superoxide dismutase 2 and peroxiredoxin 6 in asthenospermia and their correlation with sperm motility[J]. Journal of Third Military Medical University, 2021, 43(20): 2199-2207.
DOI：

Zihan YANG, Xiaoling HAO, Hong WANG, et al. Expression changes of superoxide dismutase 2 and peroxiredoxin 6 in asthenospermia and their correlation with sperm motility[J]. Journal of Third Military Medical University, 2021, 43(20): 2199-2207. DOI： 10.16016/j.1000-5404.202104129.

摘要

目的

探究活性氧(reactive oxygen species，ROS)对精子的损伤作用，分析抗氧化蛋白超氧化物歧化酶2(SOD2)和过氧化物酶6(PRDX6)在弱精症患者精子中的表达变化，并分析其与精子运动能力的相关性。

方法

选取2020年1-11月我中心收集的弱精症患者精液标本(PR

32%，PR+NP

40%，精子密度≥15×10

/mL)84例，正常精液标本(PR≥32%，PR+NP≥40%，精子密度≥15×10

/mL)55例。通过精液计算机辅助分析系统(computer-aided sperm analysis，CASA)检测精液常规参数及精子运动参数；不同浓度H

处理正常精子后通过CASA检测精子活力和运动参数变化，伊红染色检测精子存活率变化，Western blot检测精子SOD2和PRDX6表达变化；运用实时荧光定量PCR(RT-qPCR)和Western blot同时对弱精症组精子和正常组精子SOD2和PRDX6表达进行定量检测，进一步统计分析SOD2和PRDX6的表达变化及其与精液参数的相关性。

结果

CASA检测结果显示精子活力在0.10、0.35、1.00 mmol/L H

处理后显著降低；精子运动参数中曲线速率(VCL)、直线速率(VSL)、平均路径速率(VAP)和平均角位移(MAD)在0.05、0.10、0.35 mmol/L H

处理后剂量依赖式降低，侧摆幅度(ALH)、直线性(LIN)、摆动性(WOB)和前向性(STR)在0.10、0.35 mmol/L H

处理后降低；伊红染色结果显示精子存活率在0.35、1.00 mmol/L H

处理后降低，而Western blot检测结果显示精子SOD2和PRDX6表达呈H

(0.10、0.35和1.00 mmol/L)剂量依赖式增高。RT-qPCR检测结果显示弱精症组精子SOD2[(0.297±0.038)

(0.895±0.140)，

0.01

]

与PRDX6[(0.743±0.107)

(1.416±0.163)，

0.01

]

mRNA相对表达量低于正常组。Western blot结果显示弱精症组精子SOD2[(0.689±0.147)

(1.466±0.212)，

0.01

]

与PRDX6[(0.726±0.084)

(1.183±0.100)，

0.01

]

蛋白相对表达量低于正常组。相关性分析结果显示精子SOD2 mRNA相对表达量与前向运动精子百分率(

=0.267，

0.01)和总活力(

=0.329，

0.01)呈正相关；PRDX6 mRNA相对表达量与前向运动精子百分率(

=0.453，

0.01)、总活力(

=0.476，

0.01)、STR(

=0.270，

0.01)、VCL(

=0.313，

0.01)、VSL(

=0.349，

0.01)、VAP(

=0.410，

0.01)呈正相关。

结论

ROS可降低精子活力、运动能力和存活率并上调精子SOD2和PRDX6表达；弱精症患者精子SOD2和PRDX6表达下调且其表达与精子运动能力正相关。

Abstract

Objective

To explore the damaging effects of reactive oxygen species (ROS) on spermatozoa

and investigate the expression changes of superoxide dismutase 2 (SOD2) and peroxiredoxin 6 (PRDX6) in spermatozoa of asthenospermia patients

so as to analyze their correlation with sperm motility.

Methods

A total of 84 semen samples from asthenospermia patients [progressive motility (PR)

32%

PR+ non-progressive (NP)

40%

sperm density ≥15×10

/mL

]

and 55 normal semen samples (PR ≥32%

PR+NP≥40%

sperm density ≥15×10

/mL) were collected in our center from January to November 2020. The normal semen parameters were detected using a computer-aided sperm analysis (CASA) system. After the samples were treated with different concentrations of H

and the altered parameters were measured by CASA. The changes in sperm survival rate were determined by eosin staining

and the expression changes of SOD2 and PRDX6 were evaluated by Western blotting. Finally

RT-qPCR and Western blotting were adopted to compare the levels of SOD2 and PRDX6 in spermatozoa between the asthenospermia patients and normal controls

and the correlation of the levels with semen parameters were further analyzed.

Results

CASA results showed that sperm motility was significantly restrained after 0.10

0.35 and 1.00 mmol/L H

treatment; and the motility parameters like curvilinear velocity (VCL)

straight line velocity (VSL)

average path velocity (VAP) and mean angular displacement (MAD) were decreased in a dose-dependent manner with the increase of H

concentration

while the amplitude of lateral head displacement (ALH)

linearity (LIN)

wobble (WOB) and straightness (STR) were reduced at concentrations of 0.10 and 0.35 mmol/L. Eosin staining indicated that the sperm survival rate was decreased after 0.35 and 1.00 mmol/L H

treatment

and Western blotting showed the expression of SOD2 and PRDX6 in spermatozoa was up-regulated at a dose-dependent manner with the increase

of H

concentration. In addition

both the mRNA levels of SOD2 (0.297±0.038

0.895±0.140

0.01) and PRDX6 (0.743±0.107

1.416±0.163

0.01) in spermatozoa were significantly lower in the asthenospermia patients than the normal controls

and so were their protein levels (SOD: 20.689±0.147

1.466±0.212

0.01; PRDX6: 0.726±0.084

1.183±0.100

0.01). Moreover

the mRNA expression level of SOD2 was positively correlated with the sperm progressive motility (

=0.267

0.01) and total motility (

=0.329

0.01)

and that of PRDX6 was positively with sperm progressive motility (

=0.453

0.01)

total motility (

=0.476

0.01)

STR (

=0.270

0.01)

VCL (

=0.313

0.01)

VSL (

=0.349

0.01) and VAP (

=0.410

0.01).

Conclusion

ROS reduces sperm motility and survival rate

and up-regulates the expression of SOD2 and PRDX6. The down-regulated levels of SOD2 and PRDX6 are positively correlated with sperm motility in asthenospermia patients.

关键词

Keywords

references

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