Yao Wen-Liang, Liu Shao-Ming, Lin Xu-Yao, Zhou Yu-Liang, Chen Sheng-Hui, Qin Guo-Zheng
Graduate School, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China.
Department of Andrology, Hospital of Reproduction Affiliated to Jiangxi University of Chinese Medicine, Nanchang, Jiangxi 330001, China.
Zhonghua Nan Ke Xue. 2022 Jul;28(7):628-634.
To investigate the mechanism of Xianfang Huoming Decoction (XHD) improving sperm motility in mice with asthenospermia (AS).
Thirty normal BALB/c mice were randomly divided into six groups, blank control, AS model control, low-dose XHD, medium-dose XHD, high-dose XHD and levocarnitine + vitamin E (LC+VE). The AS model was established in the latter five groups by injection of methotrexate at 0.5 mg/kg once a week, and the mice in the blank control group were injected with the same volume of normal saline, all for 8 weeks. From the ninth week, the animals in the blank control and AS model control groups were treated with PBS at 0.1 ml/d, those in the low-, medium- and high-dose XHD groups with XHD at 7.13, 14,2 and 28.52 g/kg/ d respectively, and those in the LC+VE group with LC+VE (30:1) at 0.55 g/kg/d, all for 4 weeks. Then, the bilateral epididymides were harvested from all the mice for preparation of a sperm suspension and observation of the total numbers of sperm and motile sperm. The testis tissues were obtained for to determination of the expressions of Nrf-2- and HO-1-related mRNA and proteins by fluorescence staining, RT-PCR and Western blot.
Compared with the AS model controls, the mice treated with low-, medium- and high-dose XHD showed dramatically increased sperm concentration (［22.36 ± 16.02］ vs ［39.04 ± 4.50］, ［40.76 ± 6.57］ and ［41.04 ± 8.39］ ×10⁶/ml, P < 0.01) and motility (［22.89 ± 14.96］% vs ［47.98 ± 4.74］%, ［48.53 ± 6.03］% and ［49.31 ± 6.24］%, P< 0.01), decreased level of reactive oxygen species (ROS) (［16.82 ± 14.96］% vs ［12.08 ± 3.26］%, ［10.77 ± 2.21］% and ［9.56 ± 2.08］%, P< 0.01), and up-regulated expressions of Nrf-2- and HO-1-related mRNA and proteins in the testis tissue (P < 0.05 or P < 0.01).
Xianfang Huoming Decoction inhibits the development of oxidative stress by up-regulating the expressions of Nrf-2- and HO-1-related mRNA and proteins in the testis tissue, which has provided theoretical evidence for its clinical application in the treatment of asthenospermia.
探讨仙方活命饮改善弱精子症（AS）小鼠精子活力的机制。
将30只正常BALB/c小鼠随机分为6组，即空白对照组、AS模型对照组、仙方活命饮低剂量组、仙方活命饮中剂量组、仙方活命饮高剂量组和左卡尼汀+维生素E（LC+VE）组。后5组小鼠每周1次腹腔注射甲氨蝶呤0.5mg/kg建立AS模型，空白对照组小鼠注射等体积生理盐水，均连续8周。从第9周起，空白对照组和AS模型对照组小鼠每日腹腔注射0.1ml PBS，仙方活命饮低、中、高剂量组小鼠分别每日灌胃给予仙方活命饮7.13、14.2和28.52g/kg，LC+VE组小鼠每日灌胃给予LC+VE（30∶1）0.55g/kg，均连续4周。之后，采集所有小鼠双侧附睾制备精子悬液，观察精子总数及活动精子数。取睾丸组织，采用荧光染色、RT-PCR及Western blot法检测Nrf-2和HO-1相关mRNA及蛋白表达。
与AS模型对照组比较，仙方活命饮低、中、高剂量组小鼠精子浓度显著升高（［22.36±16.02］vs［39.04±4.50］、［40.76±6.57］和［41.04±8.39］×10⁶/ml, P<0.01）及精子活力显著提高（［22.89±14.96］% vs［47.98±4.74］%、［48.53±6.03］%和［49.31±6.24］%, P<0.01），活性氧（ROS）水平降低（［16.82±14.96］% vs［12.08±3.26］%、［10.77±2.21］%和［9.56±2.08］%, P<0.01），睾丸组织中Nrf-2和HO-1相关mRNA及蛋白表达上调（P<0.05或P<0.01）。
仙方活命饮通过上调睾丸组织中Nrf-2和HO-1相关mRNA及蛋白表达抑制氧化应激的发生发展，为其临床治疗弱精子症提供了理论依据。
