Wang Heng, Liu Zhaofei, Xu Xiangjun, Zhang Haitao, He Lei, Li Ming
Lianyungang Affiliated Hospital of Nanjing University of Chinese Medicine, Lianyungang, China.
Nanjing Medical University Kangda College Affiliated Hospital, Lianyungang, China.
Am J Reprod Immunol. 2025 May;93(5):e70084. doi: 10.1111/aji.70084.
Chronic prostatitis (CP), a frequent male urological disease, is featured with chronic pelvic pain and various discomfort. Calycosin has anti-inflammatory effect and its exact mechanism in CP remains unknown.
Our research was designed to explain the underlying mechanism of calycosin in CP and illustrate our findings in the rats and LPS-ATP-induced RWPE-1 cells.
The pathology of prostate tissues was analyzed by HE staining and histological inflammation score. The concentration of inflammatory factors were detected via ELISA assay. Besides, the representative biomarkers of oxidative stress were measured through detection kits. The ROS production, NLRP3 and GSDMD expression were evaluated by immunofluorescence or immumohistochemical staining. Furthermore, the expression of NF-kBp65 pathway-associated proteins and pyroptosis-associated proteins were detected by western blotting. RWPE-1 cells growth and pyroptosis were assessed by CCK-8 and flow cytometry, respectively. mRNA levels of pyroptosis markers were detected by qRT-PCR analysis.
The CP model in vivo and in vitro was successfully established by carrageenan and LPS-ATP, respectively. Calycosin substantially ameliorated the pathological damage of prostate tissue. Moreover, calycosin treatment promoted RWPE-1 cells viability and reduced pyroptosis. Calycosin remarkably reduced the inflammatory factors secretion and oxidative stress markers. Calycosin stimulation effectively suppressed the pyroptosis, which was linked with inactivation of the NF-kBp65 signaling pathway. These beneficial effects of calycosin were similar to the NLRP3 inhibitor (MCC950).
To sum up, our observations indicated that calycosin protects against CP via suppressing inflammation, oxidative stress, and pyroptosis through NF-kBp65 pathway, thereby serving as a promising therapy for CP treatment.
慢性前列腺炎（CP）是一种常见的男性泌尿系统疾病，以慢性盆腔疼痛和各种不适为特征。毛蕊异黄酮具有抗炎作用，但其在CP中的具体机制尚不清楚。
本研究旨在解释毛蕊异黄酮在CP中的潜在机制，并在大鼠和LPS-ATP诱导的RWPE-1细胞中阐述我们的研究结果。
通过HE染色和组织学炎症评分分析前列腺组织的病理学。通过ELISA法检测炎症因子的浓度。此外，通过检测试剂盒测量氧化应激的代表性生物标志物。通过免疫荧光或免疫组织化学染色评估ROS产生、NLRP3和GSDMD表达。此外，通过蛋白质印迹法检测NF-kBp65通路相关蛋白和细胞焦亡相关蛋白的表达。分别通过CCK-8和流式细胞术评估RWPE-1细胞的生长和细胞焦亡。通过qRT-PCR分析检测细胞焦亡标志物的mRNA水平。
分别通过角叉菜胶和LPS-ATP成功建立了体内和体外CP模型。毛蕊异黄酮显著改善了前列腺组织的病理损伤。此外，毛蕊异黄酮处理促进了RWPE-1细胞的活力并减少了细胞焦亡。毛蕊异黄酮显著降低了炎症因子的分泌和氧化应激标志物。毛蕊异黄酮刺激有效地抑制了细胞焦亡，这与NF-kBp65信号通路的失活有关。毛蕊异黄酮的这些有益作用与NLRP3抑制剂（MCC950）相似。
综上所述，我们的观察结果表明，毛蕊异黄酮通过NF-kBp65通路抑制炎症、氧化应激和细胞焦亡，从而对CP起到保护作用，有望成为CP治疗的一种疗法。
