Wang Sheng-Chao, Li Ben-Ke, Kan Yu-Xuan, Zeng Meng-Nan, Zhang Bei-Bei, Liu Li, Zheng Xiao-Ke, Feng Wei-Sheng
School of Pharmaceutics, Henan University of Chinese Medicine, Zhengzhou, Henan 450046, China.
Collaborative Innovation Center of Henan Province for Diagnosis and Treatment of Respiratory Diseases and Development of New Drugs, Zhengzhou, Henan 450046, China.
Zhonghua Nan Ke Xue. 2020 Jan;26(1):63-73.
To study the effect of DuzhongButiansu Capsules (DBC) on adenine-induced reproductive dysfunction (RD) in male rats.
Eighty male SD rats were randomly divided into six groups, blank control (n = 8), solvent control (n = 8), RD model control (n = 16), Shengjing Capsules (SJC) (n = 16), low-dose DBC (n = 16) and high-dose DBC (n = 16). The RD model was made by intragastric administration of adenine at 200 mg/kg/d for 5 successive weeks in the latter four groups of animals, and in the meantime the rats in the latter three groups were treated intragastrically with SJC at 0.560 mg/kg/d and DBC at 0.242 and 0.968 mg/kg/d, respectively. At the end of the fourth week, all the rats were mated with female ones in a 1:1 ratio for 7 days. Then the male rats were killed and the right epididymides collected for detection of sperm concentration and motility, and the female ones sacrificed after fed for another 2 weeks and the numbers of pregnancies and fetal rats were recorded. The heart, liver, spleen, lung, kidney, thymus, testis, epididymis and seminal vesicle were harvested for obtainment of the visceral coefficients and semen parameters, observation of the histopathological changes in the testis, epididymis and kidneys by HE staining, measurement of the levels of serum T, E2, FSH and LH by ELISA, detection of the contents of serum glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), malondialdehyde (MDA) and creatinine (Scr), blood urea nitrogen (BUN), and determination of the expressions of Bax, Bcl-2, Caspase-3 and Caspase-9 proteins in the renal tissue by immunohistochemistry.
No statistically significant difference was observed between the blank control and solvent control groups in any of the indexes obtained (P > 0.05).Compared with the blank controls, the rats in the RD model control group showed significantly decreased sperm concentration (［40.67 ± 7.37］vs ［27.10 ± 2.72］ ×10⁶/ml, P < 0.01), sperm motility (［54.75 ± 3.92］%vs ［25.60 ± 4.83］%, P < 0.01) and pregnancy rate (85.7% vs 43.8%, P < 0.01). The rats in thelow- and high-dose DBCgroups exhibited remarkable increases in sperm concentration (［53.00 ± 4.55］% and ［65.63 ± 12.47］% ×10⁶/ml, P < 0.01) and sperm motility (［53.50 ± 8.83］% and ［54.33 ± 7.92］ %, P < 0.01), and so did those in the high-dose DBC group in pregnancy rate (54.5%, P < 0.01).After medication, the animals showed markedly increased body weight and visceral coefficients of the testis, epididymis and seminal vesicle (P < 0.05 or P < 0.01), recovered morphology of the testis, epididymis and kidneys, reduced levels of Scr, BUN, FSH, LH and MDA in the serum (P < 0.05 or P < 0.01), increased contents of T, SOD and GSH-PX (P < 0.05 or P < 0.01), down-regulated expressions of Bax, Caspase-3 and Caspase-9 and up-regulated expression of Bcl-2 in the renal tissue (P < 0.05 or P < 0.01).
DBC can improve adenine-induced reproductive dysfunction in male rats, which may be attributed to its effects of inhibiting the apoptosis of proteins, improving oxidative stress and elevating the levels of reproductive hormones.
研究杜仲补天素胶囊（DBC）对腺嘌呤诱导的雄性大鼠生殖功能障碍（RD）的影响。
将80只雄性SD大鼠随机分为6组，空白对照组（n = 8）、溶剂对照组（n = 8）、RD模型对照组（n = 16）、生精胶囊（SJC）组（n = 16）、低剂量DBC组（n = 16）和高剂量DBC组（n = 16）。后四组动物连续5周以200 mg/kg/d的剂量灌胃腺嘌呤制备RD模型，同时后三组大鼠分别以0.560 mg/kg/d的剂量灌胃SJC和0.242、0.968 mg/kg/d的剂量灌胃DBC。在第4周结束时，所有大鼠与雌性大鼠按1:1的比例交配7天。然后处死雄性大鼠，收集右侧附睾检测精子浓度和活力，雌性大鼠在再喂养2周后处死，记录怀孕和胎鼠数量。摘取心脏、肝脏、脾脏、肺脏、肾脏、胸腺、睾丸、附睾和精囊，计算脏器系数和精液参数，通过苏木精-伊红（HE）染色观察睾丸、附睾和肾脏的组织病理学变化，采用酶联免疫吸附测定（ELISA）法检测血清睾酮（T）、雌二醇（E2）、卵泡刺激素（FSH）和黄体生成素（LH）水平，检测血清谷胱甘肽过氧化物酶（GSH-Px）、超氧化物歧化酶（SOD）、丙二醛（MDA）和肌酐（Scr）、血尿素氮（BUN）含量，采用免疫组织化学法检测肾组织中Bax、Bcl-2、半胱天冬酶-3（Caspase-3）和半胱天冬酶-9（Caspase-9）蛋白的表达。
空白对照组和溶剂对照组在各项检测指标上均无统计学差异（P > 0.05）。与空白对照组相比，RD模型对照组大鼠的精子浓度（［40.67 ± 7.37］vs ［27.10 ± 2.72］×10⁶/ml，P < 0.01）、精子活力（［54.75 ± 3.92］% vs ［25.60 ± 4.83］%，P < 0.01）和怀孕率（85.7% vs 43.8%，P < 0.01）显著降低。低剂量和高剂量DBC组大鼠的精子浓度（［53.00 ± 4.55］%和［65.63 ± 12.47］%×10⁶/ml，P < 0.01）和精子活力（［53.50 ± 8.83］%和［54.33 ± 7.92］%，P < 0.01）显著升高，高剂量DBC组大鼠的怀孕率也显著升高（54.5%，P < 0.01）。给药后，动物体重以及睾丸、附睾和精囊的脏器系数显著增加（P < 0.05或P < 0.01），睾丸、附睾和肾脏的形态恢复，血清Scr、BUN、FSH、LH和MDA水平降低（P < 0.05或P < 0.01）,T、SOD和GSH-PX含量增加（P < 0.05或P < 0.01），肾组织中Bax、Caspase-3和Caspase-9的表达下调，Bcl-2的表达上调（P < 0.05或P < 0.01）。
杜仲补天素胶囊可改善腺嘌呤诱导的雄性大鼠生殖功能障碍，其机制可能与其抑制蛋白凋亡、改善氧化应激及提高生殖激素水平有关。
