Xu Wenlong, Ma Wenming, Yue Jiabin, Hu Yongtao, Zhang Yi, Wang Haojie, Tai Sheng, Chen Jing, Liang Chaozhao
Department of Urology, The First Affiliated Hospital of Anhui Medical University, Institute of Urology, Anhui Medical University, and Anhui Province Key Laboratory of Urological and Andrological Diseases Research and Medical Transformation, Hefei 230022, Anhui, PR China.
Department of Urology, The Fifth Affiliated Hospital of Sun Yat-Sen University, Zhuhai 519000, Guangdong, PR China.
Phytomedicine. 2025 Jul;142:156732. doi: 10.1016/j.phymed.2025.156732. Epub 2025 Apr 10.
Juglone, a naphthoquinone compound that occurs naturally, is present predominantly in the fruits, leaves, and roots of walnut plants. Although its antioxidant and anti-inflammatory effects have been demonstrated in various diseases, its therapeutic potential remains unexplored in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS).
Our objective was to investigate the therapeutic effectiveness of juglone in treating CP/CPPS and elucidate the potential mechanism involved.
To establish experimental autoimmune prostatitis (EAP) mouse models and macrophage pyroptosis models, the therapeutic impact of juglone on CP/CPPS was evaluated. Molecular docking analysis, a cellular thermal shift assay (CETSA), and consultation with the Human Protein Atlas database were conducted to further explore the target molecules involved in juglone treatment for CP/CPPS. In addition, we utilized immunohistochemistry, immunofluorescence, Western blotting, and flow cytometry to assess macrophage pyroptosis and related pathway protein expressions. The evaluation of oxidative stress (OxS) was conducted through malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPx) assays. BzATP, an agonist of the NLRP3 pyroptosis pathway, was utilized for recovery experiments both in vitro and in vivo.
Administration of juglone to EAP model mice ameliorated prostatic inflammation, reduced pain symptoms, and decreased proinflammatory cytokine levels. Molecular docking analysis and CETSA, in conjunction with data from the Human Protein Atlas database, indicated that NLRP3, caspase-1, and GSDMD, along with their effects on macrophage pyroptosis, may serve as key targets for the effects of juglone. Furthermore, juglone inhibited the expression of these proteins. Assays of OxS demonstrated that the administration of juglone mitigated OxS in both animal and cellular experiments. These results were reversed with BzATP treatment.
In conclusion, juglone can alleviate EAP by suppressing the pyroptosis of macrophages mediated by NLRP3/GSDMD and alleviating OxS; therefore, juglone has the potential as a therapeutic for CP/CPPS. Furthermore, our studies confirmed that juglone can bind stably to NLRP3, caspase-1, and GSDMD. These findings validate the mechanism of action of juglone and offer valuable insights for the treatment of other diseases mediated by these proteins, such as inflammatory bowel disease, nonalcoholic steatohepatitis, and multiple sclerosis.
胡桃醌是一种天然存在的萘醌化合物，主要存在于核桃植物的果实、叶子和根部。尽管其抗氧化和抗炎作用已在多种疾病中得到证实，但其在慢性前列腺炎/慢性盆腔疼痛综合征（CP/CPPS）患者中的治疗潜力仍未得到探索。
我们的目的是研究胡桃醌治疗CP/CPPS的疗效，并阐明其潜在机制。
为建立实验性自身免疫性前列腺炎（EAP）小鼠模型和巨噬细胞焦亡模型，评估了胡桃醌对CP/CPPS的治疗效果。进行分子对接分析、细胞热位移分析（CETSA）并参考人类蛋白质图谱数据库，以进一步探索参与胡桃醌治疗CP/CPPS的靶分子。此外，我们利用免疫组织化学、免疫荧光、蛋白质印迹和流式细胞术来评估巨噬细胞焦亡及相关信号通路蛋白表达。通过丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GPx）检测来评估氧化应激（OxS）。NLRP3焦亡途径的激动剂BzATP用于体内外的恢复实验。
给EAP模型小鼠施用胡桃醌可改善前列腺炎症，减轻疼痛症状，并降低促炎细胞因子水平。分子对接分析和CETSA以及人类蛋白质图谱数据库的数据表明，NLRP3、半胱天冬酶-1和Gasdermin D及其对巨噬细胞焦亡的影响可能是胡桃醌发挥作用的关键靶点。此外，胡桃醌抑制了这些蛋白质的表达。氧化应激检测表明，在动物和细胞实验中，施用胡桃醌均可减轻氧化应激。BzATP处理可逆转这些结果。
总之，胡桃醌可通过抑制NLRP3/Gasdermin D介导的巨噬细胞焦亡和减轻氧化应激来缓解EAP；因此，胡桃醌有潜力用于治疗CP/CPPS。此外，我们的研究证实胡桃醌可以与NLRP3、半胱天冬酶-1和Gasdermin D稳定结合。这些发现验证了胡桃醌的作用机制，并为治疗由这些蛋白质介导的其他疾病，如炎症性肠病、非酒精性脂肪性肝炎和多发性硬化症，提供了有价值的见解。
