Department of Pediatric Surgery, Hangzhou Children's Hospital, Hangzhou, Zhejiang, China; Department of Urology, Children's Hospital of Soochow University, Suzhou, Jiangsu, 215025, China.
Department of Urology, Children's Hospital of Soochow University, Suzhou, Jiangsu, 215025, China.
Exp Cell Res. 2024 Sep 1;442(1):114218. doi: 10.1016/j.yexcr.2024.114218. Epub 2024 Aug 22.
Bladder fibrosis is the final common pathway of neurogenic bladder (NB), and its underlying mechanisms are not fully understood. The current study aims to evaluate the involvement of Piezo1, a mechanosensitive channel, in bladder fibrosis. A full-thickness bladder specimen was taken during ileocystoplasty or ureteral reimplantation from the surgical cut's edge. By chopping off the bilateral lumbar 6 (L6) and sacral 1 (S1) spinal nerves, NB rat models were produced. Utilizing both pharmacological inhibition and Piezo1 deletion, the function of Piezo1 in the TGF-β1-induced fibrosis model of SV-HUC-1 cells was delineated. RNA-seq, immunofluorescence, immunohistochemistry (IHC), and Western blotting were used to evaluate the degrees of fibrosis and biochemical signaling pathways. Piezo1 protein expression was noticeably elevated in the human NB bladder. The abundance of Piezo1 protein in bladder of NB rats was significantly increased. RNA-seq analysis revealed that the ECM-receptor interaction signaling pathway and collagen-containing ECM were increased in spinal cord injury (SCI)-induced bladder fibrosis. Moreover, the bladder of the NB rat model showed activation of YAP1 and TGF-β1/Smad. In SV-HUC-1 cells, siRNA suppression of Piezo1 led to profibrotic responses and activation of the TGF-β1/Smad pathway. However, Yoda1, a Piezo1-specific agonist, significantly reduced these effects. TGF-β1 increased Piezo1 activation and profibrotic responses in SV-HUC-1 cells. In the TGF-β1-induced fibrosis model of SV-HUC-1 cells, the TGF-β1/Smad pathway was activated, whereas the Hippo/YAP1 signal pathway was blocked. Inhibition of Piezo1 further prevented this process. Piezo1 is involved in the progression of NB bladder fibrosis and profibrotic alterations in SV-HUC-1 cells, likely through regulating the TGF-β1/Smad and Hippo/YAP1 pathways.
膀胱纤维化是神经源性膀胱（NB）的最终共同途径，其潜在机制尚未完全阐明。本研究旨在评估机械敏感通道 Piezo1 在膀胱纤维化中的作用。在回肠膀胱或输尿管再植入术中，从手术切口边缘取全层膀胱标本。通过切断双侧第 6 腰椎（L6）和第 1 骶骨（S1）脊神经，制作 NB 大鼠模型。利用药理学抑制和 Piezo1 缺失，研究了 Piezo1 在 SV-HUC-1 细胞 TGF-β1 诱导的纤维化模型中的作用。利用 RNA-seq、免疫荧光、免疫组织化学（IHC）和 Western blot 评估纤维化程度和生化信号通路。Piezo1 蛋白在人 NB 膀胱中的表达明显升高。NB 大鼠膀胱中 Piezo1 蛋白的丰度显著增加。RNA-seq 分析显示，脊髓损伤（SCI）诱导的膀胱纤维化中 ECM-受体相互作用信号通路和含胶原的 ECM 增加。此外，NB 大鼠模型的膀胱中 YAP1 和 TGF-β1/Smad 被激活。在 SV-HUC-1 细胞中，Piezo1 的 siRNA 抑制导致成纤维反应和 TGF-β1/Smad 通路的激活。然而，Piezo1 的特异性激动剂 Yoda1 显著降低了这些作用。TGF-β1 增加了 SV-HUC-1 细胞中 Piezo1 的激活和成纤维反应。在 SV-HUC-1 细胞的 TGF-β1 诱导的纤维化模型中，TGF-β1/Smad 通路被激活，而 Hippo/YAP1 信号通路被阻断。Piezo1 的抑制进一步阻止了这一过程。Piezo1 参与 NB 膀胱纤维化的进展和 SV-HUC-1 细胞的成纤维反应改变，可能通过调节 TGF-β1/Smad 和 Hippo/YAP1 通路。
