Diabetes Research Centre, Traditional Chinese Medicine School, Beijing University of Chinese Medicine, Beijing, 100029, China.
Department of Histo-embryology, Traditional Chinese Medicine School, Beijing University of Chinese Medicine, Beijing, 100029, China.
J Ethnopharmacol. 2021 May 10;271:113810. doi: 10.1016/j.jep.2021.113810. Epub 2021 Jan 26.
Sperm infertility and testicular atrophy are symptoms associated with aging. BaZiBuShen formula (BZBS), a patented Chinese herbal prescription composed of Semen Cuscutae, Fructus Lycii, Epimedii Folium, Fructus Schisandrae Sphenantherae, Fructus Cnidii, Fructus Rosae Laevigatae, Semen Allii Tuberosi., Radix Morindae Officinalis, Herba Cistanches, Fructus Rubi, Radix Rehmanniae Recens, Radix Cyathulae, Radix Ginseng, Cervi Cornu Pantotrichum, Hippocampus, and Fuctus Toosendan, has been used as a kidney-tonifying and anti-aging drug as well as for the treatment of impotence and male infertility in traditional Chinese medicine.
We aimed at investigating whether BZBS preserves sperm and testes morphology in aging mice, and to explore the underlying mechanisms.
BZBS was orally administered to aging mice induced by D-galactose (D-gal) and NaNO for 65 days. Sperm quality and testes pathophysiological alterations were examined by a Semen Analysis System, hematoxylin-eosin staining, transmission electron microscopy, and mitochondrial complex IV activity. In addition, serum levels of total antioxidant capacity (TAC), malondialdehyde (MDA), 8-hydroxy-desoxyguanosine (8-OH-dG), reduced glutathione (GSH), oxidized glutathione disulfide (GSSG), testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E) and tumor necrosis factor-α (TNF-α) were determined by ELISA. The expressions of P450 aromatase (CYP19), sirtuin 6 (Sirt6), P53, inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB)-p65, and phospho-NF-κB-p65 (NF-κB-pp65) in the testes were examined by western blot and/or immunohistochemical staining.
Sustained exposure to D-gal/NaNO caused a deterioration of sperm quality and testes morphology in this rapid aging mouse model. BZBS treatment curtailed these alterations. These beneficial effects were associated with increased serum levels of TAC, GSH/GSSG, T, E, and FSH, and decreased levels of MDA, TNF-α, and 8-OH-dG. BZBS treatment also downregulated the expressions of P53, iNOS, and NF-κB-pp65, as well as upregulated the expressions of Sirt6 and CYP19 in aging testes.
BZBS preserves testicular morphology and spermatogenesis possibly via inhibition of oxidative stress and the modulation of the Sirt6/P53 and Sirt6/NF-κB signaling pathways. The results shed light on the beneficial effect of BZBS on sperm quality and fertility in aging males.
精子不育和睾丸萎缩是与衰老相关的症状。八子补肾方（BZBS）是一种专利中药方剂，由菟丝子、枸杞子、淫羊藿、五味子、蛇床子、玫瑰花、韭菜子、肉苁蓉、淫羊藿、肉苁蓉、覆盆子、熟地黄、牛膝、人参、鹿茸、海马和苦杏仁组成，已被用作补肾和抗衰老药物，以及治疗中医的阳痿和男性不育症。
我们旨在研究 BZBS 是否能保存衰老小鼠的精子和睾丸形态，并探讨其潜在机制。
用 D-半乳糖（D-gal）和 NaNO 诱导衰老小鼠 65 天，用 BZBS 灌胃。通过精子分析系统、苏木精-伊红染色、透射电镜和线粒体复合物 IV 活性检查精子质量和睾丸病理生理改变。此外，通过 ELISA 测定血清总抗氧化能力（TAC）、丙二醛（MDA）、8-羟基脱氧鸟苷（8-OH-dG）、还原型谷胱甘肽（GSH）、氧化型谷胱甘肽二硫化物（GSSG）、睾酮（T）、卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E）和肿瘤坏死因子-α（TNF-α）的水平。通过 Western blot 和/或免疫组化染色检测睾丸中 P450 芳香酶（CYP19）、Sirtuin 6（Sirt6）、P53、诱导型一氧化氮合酶（iNOS）、核因子-κB（NF-κB）-p65 和磷酸化-NF-κB-p65（NF-κB-pp65）的表达。
持续暴露于 D-gal/NaNO 在这种快速衰老小鼠模型中导致精子质量和睾丸形态恶化。BZBS 治疗减轻了这些改变。这些有益的作用与血清 TAC、GSH/GSSG、T、E 和 FSH 水平的升高以及 MDA、TNF-α 和 8-OH-dG 水平的降低有关。BZBS 治疗还下调了衰老睾丸中 P53、iNOS 和 NF-κB-pp65 的表达，并上调了 Sirt6 和 CYP19 的表达。
BZBS 通过抑制氧化应激和调节 Sirt6/P53 和 Sirt6/NF-κB 信号通路来保存睾丸形态和精子发生。结果揭示了 BZBS 对衰老雄性精子质量和生育能力的有益影响。
