Dong Yang, Zheng Yanfei, Zhu Linghui, Li Tianxing, Guan Yuanyuan, Zhao Shipeng, Wang Qi, Wang Ji, Li Lingru
School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China.
National Institute of TCM Constitution and Preventive Medicine, Beijing University of Chinese Medicine, Beijing, China.
Front Pharmacol. 2022 Apr 26;13:896434. doi: 10.3389/fphar.2022.896434. eCollection 2022.
Oligoasthenozoospermia is the leading cause of male infertility, seriously affecting men's health and increasing the societal medical burden. In recent years, obesity-related oligoasthenozoospermia has attracted increased attention from researchers to find a cure. This study aimed to evaluate the efficacy of Hua-Tan-Sheng-Jing decoction (HTSJD) in treating obesity with oligoasthenozoospermia, determine its active ingredients and identify its mechanism of action. The ingredients of HTSJD were determined by combining the ultra-performance liquid chromatography with mass spectrometry (UPLC-MS/MS) and systems pharmacology approach. The common pathogenesis of obesity and oligoasthenozoospermia and the potential mechanism of HTSJD against obesity with oligoasthenozoospermia were obtained through target fishing, network construction, and enrichment analyses. Further, molecular docking of the key ingredients with the upstream receptors of the key signaling pathways of the potential mechanism was used to predict their affinity. Finally, high-fat-induced obesity with oligoasthenozoospermia rat model was constructed to determine the effects of HTSJD on semen concentration, sperm motility, body weight, and serum lipid metabolism. The key proteins were validated by immunohistochemistry (IHC). A total of 70 effective components and 847 potential targets of HTSJD (H targets) were identified, of which 743 were common targets related to obesity and oligoasthenozoospermia (O-O targets) mainly enriched in the pathways related to inflammation, oxidative stress and hormone regulation. Finally, 143 common targets (H-O-O targets) for HTSJD against obesity with oligoasthenozoospermia were obtained. Combining the hub genes and the results of Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of H-O-O targets, PI3K-AKT and MAPK signaling pathways were identified as the key pathways. Molecular docking results showed that Diosgenin, Kaempferol, Quercetin, Hederagenin, Isorhamnetin may act on the related pathways by docking EGFR, IGF1R and INSR. The animal-based experiments confirmed that HTSJD improves the sperm quality of high-fat diet-fed rats by reducing their body weight and blood lipid levels, influencing the PI3K-AKT and MAPK signaling pathways and altering the corresponding protein expressions. HTSJD treats obesity with oligoasthenozoospermia by up-regulating the PI3K-AKT signaling pathway and down-regulating the MAPK signaling pathway, which are at the crossroad of obesity and oligoasthenozoospermia.
少弱精子症是男性不育的主要原因，严重影响男性健康并增加社会医疗负担。近年来，肥胖相关的少弱精子症引起了研究人员越来越多的关注，以寻求治愈方法。本研究旨在评估化痰生精汤（HTSJD）治疗肥胖伴少弱精子症的疗效，确定其活性成分并明确其作用机制。通过超高效液相色谱-质谱联用（UPLC-MS/MS）和系统药理学方法确定了HTSJD的成分。通过靶点筛选、网络构建和富集分析，获得了肥胖和少弱精子症的共同发病机制以及HTSJD治疗肥胖伴少弱精子症的潜在机制。此外，将关键成分与潜在机制关键信号通路的上游受体进行分子对接，以预测它们之间的亲和力。最后，构建高脂诱导的肥胖伴少弱精子症大鼠模型，以确定HTSJD对精液浓度、精子活力、体重和血清脂质代谢的影响。通过免疫组织化学（IHC）验证关键蛋白。共鉴定出HTSJD的70种有效成分和847个潜在靶点（H靶点），其中743个是与肥胖和少弱精子症相关的共同靶点（O-O靶点），主要富集在与炎症、氧化应激和激素调节相关的通路中。最终，获得了HTSJD治疗肥胖伴少弱精子症的143个共同靶点（H-O-O靶点）。结合核心基因以及H-O-O靶点的基因本体（GO）功能和京都基因与基因组百科全书（KEGG）通路分析结果，确定PI3K-AKT和MAPK信号通路为关键通路。分子对接结果表明，薯蓣皂苷元、山柰酚、槲皮素、常春藤皂苷元、异鼠李素可能通过与表皮生长因子受体（EGFR）、胰岛素样生长因子1受体（IGF1R）和胰岛素受体（INSR）对接作用于相关通路。动物实验证实，HTSJD通过降低高脂饮食喂养大鼠的体重和血脂水平，影响PI3K-AKT和MAPK信号通路并改变相应蛋白表达，从而改善其精子质量。HTSJD通过上调PI3K-AKT信号通路和下调MAPK信号通路来治疗肥胖伴少弱精子症，这两条信号通路处于肥胖和少弱精子症的交叉点。
