Li Ming, Yao Wenping, Sun Yingying, Wang Heng
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 Aug;94(2):e70113. doi: 10.1111/aji.70113.
Chronic prostatitis (CP) is a common and serious disorder characterized by unknown pathogenic mechanisms and recurrent symptoms. Exosomes isolated from prostate stromal cells can also be used to treat chronic inflammation. This study aimed to elucidate the mechanism of action of Calycosin (CA) during CP therapy.
LPS volume of 10 µg/mL was applied for constructing the cell models of CP. RWPE-1 cells were co-cultivated with exosomes isolated from CA elicited-WPMY-1 cells (CA-WPMY-1-exo). Exosomes isolated from WPMY-1 cells were identified by TEM, NTA, and western blotting. MTT and flow cytometry analyses were performed to evaluate cell viability and apoptosis, respectively. The secretion of inflammatory cytokines was measured using ELISA. The expressions of cleaved-Caspase3, Caspase3, p-p65, and p65 were determined by western blotting.
LPS treatment decreased RWPE-1 cell viability and stimulated more apoptotic cells, which was partly abolished by CA treatment in a dose-dependent manner. Furthermore, CA alleviated the LPS-induced inflammatory response in a dose-dependent manner. NC-WPMY-1 and CA-WPMY-1 markedly enhanced RWPE-1 cell viability. We also found that both NC-WPMY-1-exo and CA-WPMY-1-exo reversed the effects of LPS on RWPE-1 cell viability, apoptosis, and inflammation. The effect of CA-WPMY-1-exo on RWPE-1 cells was more significant than that of NC-WPMY-1-exo.
Exosomes derived from CA-exposed prostate stromal cells were identified as significant mediators of CP by inhibiting LPS-induced epithelial cells inflammatory injury.
慢性前列腺炎（CP）是一种常见且严重的疾病，其致病机制不明且症状反复。从前列腺基质细胞中分离出的外泌体也可用于治疗慢性炎症。本研究旨在阐明毛蕊异黄酮（CA）在CP治疗过程中的作用机制。
应用10μg/mL的脂多糖构建CP细胞模型。将RWPE-1细胞与从经CA诱导的WPMY-1细胞中分离出的外泌体（CA-WPMY-1-exo）共培养。通过透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和蛋白质印迹法对外泌体进行鉴定。分别采用MTT法和流式细胞术分析评估细胞活力和凋亡情况。使用酶联免疫吸附测定（ELISA）法检测炎性细胞因子的分泌。通过蛋白质印迹法测定裂解型半胱天冬酶3（cleaved-Caspase3）、半胱天冬酶3（Caspase3）、磷酸化p65（p-p65）和p65的表达。
脂多糖处理降低了RWPE-1细胞活力并刺激更多细胞凋亡，而CA处理以剂量依赖方式部分消除了这种作用。此外，CA以剂量依赖方式减轻了脂多糖诱导的炎症反应。正常对照WPMY-1（NC-WPMY-1）和经CA诱导的WPMY-1（CA-WPMY-1）显著提高了RWPE-1细胞活力。我们还发现，NC-WPMY-1-exo和CA-WPMY-1-exo均逆转了脂多糖对RWPE-1细胞活力、凋亡和炎症的影响。CA-WPMY-1-exo对RWPE-1细胞的作用比NC-WPMY-1-exo更显著。
通过抑制脂多糖诱导的上皮细胞炎症损伤，从经CA处理的前列腺基质细胞中获得的外泌体被确定为CP的重要介质。
