Li Mingzhu, Chen Linhuang, Xu Haotian, Li Junlin, Liu Yatian, Chen Xiuyun, Luo Minyi, Xie Xinyuan, Yin Mingyu, He Jinyang
Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
Artemisinin Research Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
Pharmaceuticals (Basel). 2025 Sep 12;18(9):1363. doi: 10.3390/ph18091363.
: Oligoasthenozoospermia (OA) is a common cause of male infertility. Modified Liuwei Dihuang Decoction (MLWDH) is an improved version of Liuwei Dihuang Decoction (LWDH), a traditional Chinese medicine prescription, which has demonstrated significant therapeutic effects against OA. This study aims to evaluate the protective effects of MLWDH against OA and elucidate its underlying molecular mechanisms. : The constituents of MLWDH were identified via UPLC-HRMS and compound databases (TCMSP, HERB). Network pharmacology analysis was conducted to predict potential therapeutic targets and associated signaling pathways. In vivo, a CP-induced mouse model of OA was established to evaluate the therapeutic efficacy of MWDH by assessing testicular and epididymal indices, sperm quality, histopathological changes and serum hormone levels. Oxidative stress markers, including MDA, SOD, GSH and NO, were measured using commercial assay kits. The underlying molecular mechanisms, particularly those related to oxidative stress and inflammation (PI3K, Akt, Nrf2, Keap1, HO-1, NQO1, NF-κB, TNF-α, IL-6), were further elucidated by RT-qPCR, Western blot, and immunofluorescence. : A total of 345 major bioactive compounds were identified in MLWDH. Network pharmacology and molecular docking analyses indicated that MLWDH exerts its effects primarily through the PI3K/AKT signaling pathway. MLWDH administration in vivo significantly improved sperm count, motility, and morphology, while also increasing serum levels of testosterone, FSH, and LH. Moreover, MLWDH significantly mitigated oxidative damage, as evidenced by decreased MDA concentrations and elevated levels of GSH, NO and SOD. Mechanistic investigations further substantiated that MLWDH enhanced PI3K/AKT/Nrf2 signaling while inhibiting NF-κB signaling in OA mice. : Our findings suggest that MLWDH ameliorates OA in a preclinical mouse model by improving sperm quality and testicular function, potentially via activation of the PI3K/AKT/Nrf2 signaling pathway and the inhibition of NF-κB signaling, thereby alleviating oxidative stress and inflammatory responses.
少弱精子症（OA）是男性不育的常见原因。改良六味地黄汤（MLWDH）是中药方剂六味地黄汤（LWDH）的改良版本，已显示出对OA有显著治疗效果。本研究旨在评估MLWDH对OA的保护作用，并阐明其潜在的分子机制。：通过超高效液相色谱-高分辨质谱联用（UPLC-HRMS）和化合物数据库（中药系统药理学数据库（TCMSP）、中药综合数据库（HERB））鉴定MLWDH的成分。进行网络药理学分析以预测潜在的治疗靶点和相关信号通路。在体内，建立环磷酰胺（CP）诱导的OA小鼠模型，通过评估睾丸和附睾指数、精子质量、组织病理学变化和血清激素水平来评价MLWDH的治疗效果。使用商业检测试剂盒测量氧化应激标志物，包括丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）和一氧化氮（NO）。通过实时定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法（Western blot）和免疫荧光进一步阐明潜在的分子机制，特别是与氧化应激和炎症相关的机制（磷脂酰肌醇-3激酶（PI3K）、蛋白激酶B（Akt）、核因子E2相关因子2（Nrf2）、 Kelch样环氧氯丙烷相关蛋白1（Keap1）、血红素加氧酶-1（HO-1）、醌氧化还原酶1（NQO1）、核因子κB（NF-κB）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6））。：在MLWDH中总共鉴定出345种主要生物活性化合物。网络药理学和分子对接分析表明，MLWDH主要通过PI3K/AKT信号通路发挥作用。在体内给予MLWDH可显著改善精子数量、活力和形态，同时还可提高血清睾酮、卵泡刺激素（FSH）和黄体生成素（LH）水平。此外，MLWDH显著减轻氧化损伤，表现为MDA浓度降低以及GSH、NO和SOD水平升高。机制研究进一步证实，MLWDH在OA小鼠中增强了PI3K/AKT/Nrf2信号通路，同时抑制了NF-κB信号通路。：我们的研究结果表明，MLWDH在临床前小鼠模型中通过改善精子质量和睾丸功能来改善OA，可能是通过激活PI3K/AKT/Nrf2信号通路和抑制NF-κB信号通路，从而减轻氧化应激和炎症反应。
