Wang Wen, Han Ling, Liang Yichun, Liang Shulin, Qin Zhan, Geng Liguo, He Chaoba, Huang Ting, Yuan Shaoying
Second Clinical Medical College, Guangzhou University of CM, Guangzhou 510006, Guangdong Province, China.
Province-Ministry Co-constructing State Key Laboratory of Dampness Syndrome of TCM, Second Affiliated Hospital of Guangzhou University of CM; Research Team of Bio-molecular and System Biology of TCM, Guangdong Academy of Chinese Medical Sciences; Guangdong Provincial Key Laboratory of Clinical Research on TCM Syndrome; Guangdong-Hong Kong-Macau Joint Lab on TCM and Immune Disease Research.
Zhongguo Zhen Jiu. 2025 Apr 12;45(4):495-504. doi: 10.13703/j.0255-2930.20240105-k0003. Epub 2025 Feb 26.
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.
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·d) 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" (SP6) 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.
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).
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.
观察不同频率电针对少弱精子症(OAT)大鼠生精功能、睾丸形态及氧化应激的影响,探讨电针治疗OAT的作用机制及最佳参数。
将60只SPF级雄性SD大鼠随机分为溶剂对照组、模型组、2Hz电针组、100Hz电针组和2Hz/100Hz电针组,每组12只。除溶剂对照组外,其余4组大鼠采用灌胃给予奥硝唑混悬液(800mg·kg·d),连续28天,建立OAT模型。从建模第1天开始,对3个电针组大鼠于“关元”(CV4)、“气海”(CV6)及双侧“三阴交”(SP6)、“足三里”(ST36)穴施加电针,2Hz电针组、100Hz电针组和2Hz/100Hz电针组分别采用2Hz连续波、100Hz连续波、2Hz/100Hz疏密波,电流强度1~3mA,每次30分钟,隔日1次,连续28天。干预后,计算睾丸指数,评估附睾精子质量,观察生育能力;采用HE染色观察睾丸组织形态,并计算Johnson评分;采用免疫荧光法检测睾丸组织中活性氧(ROS)的阳性表达;采用ELISA法检测睾丸组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及丙二醛(MDA)水平;采用Western blot法检测睾丸组织中核因子E2相关因子2(Nrf2)和血红素加氧酶-1(HO-1)蛋白表达。
与溶剂对照组比较,模型组大鼠睾丸指数、精子浓度、精子活力及子代数量均降低(<0.01),生精小管萎缩,Johnson评分降低(<0.01);睾丸组织中SOD、CAT活性及Nrf2、HO-1蛋白表达均降低(<0.01);精子畸形率、睾丸组织中ROS阳性表达及MDA水平升高(<0.01)。与模型组比较,2Hz电针组、100Hz电针组和2Hz/100Hz电针组大鼠睾丸指数、精子浓度、精子活力及子代数量均升高(<0.05,<0.01),睾丸组织病理形态改善,Johnson评分升高(<0.01);睾丸组织中SOD、CAT活性及Nrf2、HO-1蛋白表达均升高(<0.05,<0.01);精子畸形率、睾丸组织中ROS阳性表达及MDA水平降低(<0.05,<0.01)。与2Hz电针组比较,2Hz/第100Hz电针组大鼠睾丸指数、精子浓度、精子活力及睾丸组织中CAT活性、HO-1蛋白表达均升高(<0.01,<0.05);睾丸组织中ROS阳性表达降低(<0.01)。与100Hz电针组比较,2Hz/100Hz电针组大鼠睾丸指数升高(<0.01),睾丸组织中ROS阳性表达降低(<0.01)。
2Hz连续波、100Hz连续波及2Hz/100Hz疏密波电针均能改善OAT大鼠生精阻滞,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2Hz/100Hz疏密波电针效果最佳。