Effects of electroacupuncture with different frequencies on spermatogenesis and oxidative stress in oligoasthenospermia rats

WANG Wen; HAN Ling; LIANG Yichun; LIANG Shulin; QIN Zhan; GENG Liguo; HE Chaoba; HUANG Ting; YUAN Shaoying

doi:10.13703/j.0255-2930.20240105-k0003

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Effects of electroacupuncture with different frequencies on spermatogenesis and oxidative stress in oligoasthenospermia rats

Study on Mechanism | 更新时间：2025-04-11

- Effects of electroacupuncture with different frequencies on spermatogenesis and oxidative stress in oligoasthenospermia rats
- Chinese Acupuncture & Moxibustion Vol. 45, Issue 4, Pages: 495-504(2025)
- 作者机构：
  
  广州中医药大学第二临床医学院，广东广州　510006
  省部共建中医湿证国家重点实验室/广州中医药大学第二附属医院
  广东省中医药科学院中医药分子生物与系统生物学团队
  广东省中医证候临床研究重点实验室
  粤港澳中医药与免疫疾病研究联合实验室
  广东省中医院珠海医院男科，珠海　510915
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20240105-k0003
  CLC：
- Received：05 January 2024，
  
  Revised：2025-02-18，
  
  Online First：26 February 2025，
  
  Published：12 April 2025
- 稿件说明：
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WANG Wen, HAN Ling, LIANG Yichun, et al. Effects of electroacupuncture with different frequencies on spermatogenesis and oxidative stress in oligoasthenospermia rats[J]. Chinese Acupuncture & Moxibustion, 2025, 45(4): 495-504.
DOI：

WANG Wen, HAN Ling, LIANG Yichun, et al. Effects of electroacupuncture with different frequencies on spermatogenesis and oxidative stress in oligoasthenospermia rats[J]. Chinese Acupuncture & Moxibustion, 2025, 45(4): 495-504. DOI： 10.13703/j.0255-2930.20240105-k0003.

摘要

目的：

观察不同频率电针对少弱精子症（OAT）模型大鼠生精功能、睾丸形态及氧化应激的影响，探索电针干预OAT的作用机制及最优参数。

方法：

将60只SPF级雄性SD大鼠随机分为溶剂对照组、模型组、 2 Hz电针组、100 Hz电针组和2 Hz/100 Hz电针组，每组12只。除溶剂对照组外，其余4组大鼠以奥硝唑混悬液（800 mg·kg

-1

·d

-1

）灌胃28 d的方法制备OAT模型。从造模第1天开始，3个电针组均于“关元”“气海”及双侧“三阴交”“足三里”行电针干预，2 Hz电针组、100 Hz电针组、2 Hz/100 Hz电针组分别选择2 Hz连续波、100 Hz连续波和2 Hz/100 Hz疏密波，电流1～3 mA，每次30 min，隔日1次，连续28 d。干预后，计算各组大鼠睾丸指数，检测附睾精子质量，观察大鼠生育能力；HE染色法观察大鼠睾丸组织形态并计算Johnson评

分；免疫荧光法检测大鼠睾丸组织活性氧（ROS）阳性表达；ELISA法检测大鼠睾丸组织氧化应激指标超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性及丙二醛（MDA）含量；Western blot法检测大鼠睾丸组织核因子红细胞2相关因子2（Nrf2）及血红素加氧酶-1（HO-1）蛋白表达。

结果：

与溶剂对照组比较，模型组大鼠睾丸指数、精子浓度、精子活力及产仔数降低（

0.01）；生精小管萎缩，Johnson评分降低（

0.01）；睾丸组织SOD及CAT活性、Nrf2及HO-1蛋白表达降低（

0.01）；精子畸形率及睾丸组织ROS阳性表达、MDA含量升高（

0.01）。与模型组比较，2 Hz电针组、100 Hz电针组及2 Hz/100 Hz电针组大鼠睾丸指数、精子浓度、精子活力及产仔数升高（

0.05，

0.01）；睾丸组织病理形态改善，Johnson评分升高（

0.01）；睾丸组织SOD及CAT活性、Nrf2及HO-1蛋白表达升高（

0.05，

0.01）；精子畸形率及睾丸组织ROS阳性表达、MDA含量降低（

0.05，

0.01）。与2 Hz电针组比较，2 Hz/100 Hz电针组大鼠睾丸指数、精子浓度、精子活力及睾丸组织CAT活性、HO-1蛋白表达升高（

0.01，

0.05），睾丸组织ROS阳性表达降低（

0.01）；与100 Hz电针组比较，2 Hz/100 Hz电针组大鼠睾丸指数升高（

0.01），睾丸组织ROS阳性表达降低（

0.01）。

结论：

2 Hz低频连续波、100 Hz高频连续波、2 Hz/100 Hz疏密波电针均可改善OAT模型大鼠生精功能障碍，降低睾丸组织氧化应激水平，其机制可能与上调Nrf2、HO-1蛋白表达，改善氧化应激有关，其中2 Hz/100 Hz疏密波电针效果最优。

Abstract

Objective

To observe the effects of electroacupuncture (EA) with different frequencies on spermatogenic function

testicular morphology and oxidative stress in oligoasthenospermia (OAT) rats

and to explore the mechanism and the optimal parameters of EA for OAT.

Methods

Sixty SPF-grade male SD rats were randomly divided into a solvent control group

a model group

a 2 Hz EA group

a 100 Hz EA group and a 2 Hz/100 Hz EA group

with 12 rats in each group. Except for the solvent control group

the other 4 groups were administered ornidazole suspension (800 mg·kg

-1

·d

-1

) by gavage for 28 d to establish the OAT model. Starting from the 1st of modeling

EA was applied at "Guanyuan" (CV4)

"Qihai" (CV6) and bilateral "Sanyinjiao" (S

P6) and "Zusanli" (ST36) in the 3 EA groups

continuous wave of 2 Hz

continuous wave of 100 Hz

and disperse-dense wave of 2 Hz/100 Hz were used in the 2 Hz EA group

the 100 Hz EA group

and the 2 Hz/100 Hz EA group

respectively

with current intensity of 1-3 mA

30 min a time

once every other day

for 28 consecutive days. After intervention

the testicular index was calculated

epididymal sperm quality was assessed

and the fertility ability was observed; morphology of testicular tissue was observed by HE staining

and the Johnson score was calculated; the positive expression of reactive oxygen species (ROS) in testicular tissue was detected by immunofluorescence; the activity of superoxide dismutase (SOD) and catalase (CAT)

as well as the level of malondialdehyde (MDA) in testicular tissue were measured by ELISA; the protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in testicular tissue was detected by Western blot.

Results

Compared with the solvent control group

in the model group

the testicular index

sperm concentration

sperm motility and the number of offspring were decreased (

0.01)

the seminiferous tubules atrophied and the Johnson score decreased (

0.01); the activity of SOD and CAT

as well as the protein expression of Nrf2 and HO-1 in testicular tissue were decreased (

0.01); the sperm deformity rate

the positive expression of ROS and the MDA level in testicular tissue were increased (

0.01). Compared with the model group

in the 2 Hz EA group

the 100 Hz EA group and the 2 Hz/100 Hz EA group

the testicular index

sperm concentration

sperm motility and the number of offspring were increased (

0.05

0.01)

the pathological morphology of testicular tissue improved and the Johnson scores increased (

0.01); the activity of SOD and CAT

as well as the protein expression

of Nrf2 and HO-1 in testicular tissue were increased (

0.05

0.01); the sperm deformity rate

the positive expression of ROS and the MDA level in testicular tissue were decreased (

0.05

0.01). Compared with the 2 Hz EA group

in the 2 Hz/100 Hz EA group

the testicular index

sperm concentration

sperm motility

as well as the CAT activity and HO-1 protein expression in testicular tissue were increased (

0.01

0.05); the positive expression of ROS was decreased (

0.01). Compared with the 100 Hz EA group

in the 2 Hz/100 Hz EA group

the testicular index was increased (

0.01)

the positive expression of ROS in testicular tissue was decreased (

0.01).

Conclusion

EA with 2 Hz continuous wave

100 Hz continuous wave

and 2 Hz/100 Hz disperse-dense wave can all improve the spermatogenic arrest and reduce the level of oxidative stress in testicular tissue in OAT rats

the mechanism may be related to up-regulating the protein expression of Nrf2 and HO-1 and improving oxidative stress. EA with disperse-dense wave of 2 Hz/100 Hz shows the optimal effect.

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