Ke Hanwei, Zhu Lin, Wang Qi, Xu Kexin
Department of Urology, Peking University People's Hospital, Beijing, 100044, China.
Peking University Applied Lithotripsy Institute, Peking University People's Hospital, Beijing, 10034, China.
World J Urol. 2025 Jan 4;43(1):60. doi: 10.1007/s00345-024-05429-9.
This study aims to elucidate the role of pituitary adenylate cyclase-activating polypeptide (PACAP) in Hunner-type Interstitial Cystitis (HIC) and evaluate its potential as a therapeutic target.
Bladder tissue samples were obtained from HIC patients and normal bladder tissue from bladder cancer patients. PACAP expression was assessed through immunohistochemistry. An in vitro HIC model was established using LPS-induced SV-HUC1 cells. PACAP knockdown was performed using siRNA. The expression of inflammatory markers (IL-6, IL-1β, TNF-α) and fibrotic markers (fibronectin 1, TGF-β1, collagen I) was evaluated via qPCR, Western blot, and ELISA. Cell migration and proliferation were analyzed using wound healing and CCK-8 assays. Transcriptomic profiling was conducted to identify differentially expressed genes (DEGs) and explore their functional significance.
PACAP expression was significantly elevated in the bladder tissues of HIC patients. LPS stimulation of SV-HUC1 cells induced PACAP expression alongside increased levels of inflammatory cytokines, validating the inflammatory model. PACAP knockdown markedly suppressed IL-6, IL-1β, and TNF-α expression and attenuated LPS-induced fibrosis by reducing fibronectin 1, TGF-β1, and collagen I levels. Additionally, PACAP knockdown inhibited LPS-induced cell migration and proliferation, as evidenced by wound healing and CCK-8 assays. Transcriptomic analysis revealed distinct molecular alterations in HIC tissues, including PACAP upregulation, implicating it in HIC pathogenesis.
PACAP plays a pivotal role in the inflammatory and fibrotic pathways of HIC. PACAP knockdown alleviates LPS-induced pathological responses, highlighting its potential as a novel therapeutic target. Further research is warranted to investigate PACAP's precise mechanisms in HIC and its translational application in clinical settings.
本研究旨在阐明垂体腺苷酸环化酶激活多肽（PACAP）在Hunner型间质性膀胱炎（HIC）中的作用，并评估其作为治疗靶点的潜力。
从HIC患者获取膀胱组织样本，从膀胱癌患者获取正常膀胱组织。通过免疫组织化学评估PACAP表达。使用脂多糖（LPS）诱导的SV-HUC1细胞建立体外HIC模型。使用小干扰RNA（siRNA）进行PACAP基因敲低。通过定量聚合酶链反应（qPCR）、蛋白质免疫印迹法和酶联免疫吸附测定（ELISA）评估炎症标志物（白细胞介素6、白细胞介素1β、肿瘤坏死因子α）和纤维化标志物（纤连蛋白1、转化生长因子β1、I型胶原）的表达。使用伤口愈合实验和细胞计数试剂盒-8（CCK-8）实验分析细胞迁移和增殖。进行转录组分析以鉴定差异表达基因（DEGs）并探索其功能意义。
HIC患者膀胱组织中PACAP表达显著升高。LPS刺激SV-HUC1细胞诱导PACAP表达，同时炎症细胞因子水平升高，验证了炎症模型。PACAP基因敲低显著抑制白细胞介素6、白细胞介素1β和肿瘤坏死因子α的表达，并通过降低纤连蛋白1、转化生长因子β1和I型胶原水平减轻LPS诱导的纤维化。此外，伤口愈合实验和CCK-8实验表明，PACAP基因敲低抑制LPS诱导的细胞迁移和增殖。转录组分析揭示了HIC组织中不同的分子改变，包括PACAP上调，提示其参与HIC发病机制。
PACAP在HIC的炎症和纤维化途径中起关键作用。PACAP基因敲低减轻LPS诱导的病理反应，突出了其作为新型治疗靶点的潜力。有必要进一步研究PACAP在HIC中的精确机制及其在临床环境中的转化应用。
