The prostate is a vital accessory gonad in the mammalian male reproductive system. With the ever-increasing proportion of the population over 60 years of age worldwide, the incidence of prostate diseases, such as benign prostatic hyperplasia (BPH) and prostate cancer (PCa), is on the rise and is gradually becoming a significant medical problem globally. The notch signaling pathway is essential in regulating prostate early development. However, the potential regulatory mechanism of Notch signaling in prostatic enlargement and hyperplasia remains unclear. In this study, we proved that overactivation of Notch1 signaling in mouse prostatic epithelial cells (OEx) led to prostatic enlargement via enhancing proliferation and inhibiting apoptosis of prostatic epithelial cells. Further study showed that N1ICD/RBPJ directly up-regulated the androgen receptor (AR) and enhanced prostatic sensitivity to androgens. Hyper-proliferation was not found in orchidectomized OEx mice without androgen supply but was observed after Dihydrotestosterone (DHT) supplementation. Our data showed that the number of mitochondrion in prostatic epithelial cells of OEx mice was increased, but the mitochondrial function was impaired, and the essential activity of the mitochondrial respiratory electron transport chain was significantly weakened. Disordered mitochondrial number and metabolic function further resulted in excessive accumulation of reactive oxygen species (ROS). Importantly, anti-oxidant N-Acetyl-L-Cysteine (NAC) therapy could alleviate prostatic hyperplasia caused by the over-activation of Notch1 signaling. Furthermore, we observed the incremental Notch signaling activity in progenitor-like club cells in the scRNA-seq data set of human BPH patients. Moreover, the increased number of TROP2⁺ progenitors and Club cells was also confirmed in our OEx mice. In conclusion, our study revealed that over-activated Notch1 signaling induces prostatic enlargement by increasing androgen receptor sensitivity, disrupting cellular mitochondrial metabolism, increasing ROS, and a higher number of progenitor cells, all of which can be effectively rescued by NAC treatment.

前列腺是哺乳动物雄性生殖系统中重要的附属性腺。随着全球60岁以上人口比例不断增加，良性前列腺增生（BPH）、前列腺癌（PCa）等前列腺疾病的发病率呈上升趋势，并逐渐成为全球重大的医疗问题。 Notch信号通路对于调节前列腺早期发育至关重要。然而，Notch信号在前列腺肥大和增生中的潜在调节机制仍不清楚。在这项研究中，我们证明了小鼠前列腺上皮细胞（OEx）中Notch1信号的过度激活通过增强前列腺上皮细胞的增殖和抑制前列腺上皮细胞的凋亡而导致前列腺肥大。进一步研究表明，N1ICD/RBPJ直接上调雄激素受体（AR）并增强前列腺对雄激素的敏感性。在没有雄激素供应的睾丸切除OEx小鼠中没有发现过度增殖，但在补充二氢睾酮 (DHT) 后观察到过度增殖。我们的数据显示， OEx小鼠前列腺上皮细胞中线粒体数量增加，但线粒体功能受损，线粒体呼吸电子传递链的必需活性显着减弱。线粒体数量和代谢功能紊乱进一步导致活性氧（ROS）过度积累。重要的是，抗氧化剂 N-乙酰基-L-半胱氨酸 (NAC) 疗法可以缓解由 Notch1 信号过度激活引起的前列腺增生。此外，我们在人类 BPH 患者的 scRNA-seq 数据集中观察到类祖细胞俱乐部细胞中 Notch 信号活性的增加。此外，在我们的OEx小鼠中也证实了TROP2 ⁺祖细胞和 Club 细胞数量的增加。总之，我们的研究表明，过度激活的 Notch1 信号传导通过增加雄激素受体敏感性、破坏细胞线粒体代谢、增加 ROS 和更多祖细胞数量来诱导前列腺肥大，所有这些都可以通过 NAC 治疗有效挽救。