Wang Xiaoya, Li Ling, Sun Bishao, Hou Xianglin, Song Siqi, Shi Chunying, Chen Wei
Department of Human Anatomy, Histology and Embryology, School of Basic Medicine, Qingdao University, Qingdao, Shandong Province, China.
Department of Urology, Xinqiao Hospital of Army Medical University, Chongqing, China.
Curr Stem Cell Res Ther. 2024;19(1):103-115. doi: 10.2174/1574888X18666230331123540.
Urine-derived stem cells (USCs) were considered to be an ideal source of stem cells for repairing urological diseases. However, the proliferative ability of USCs significantly decreased when cultured on plastic dishes, which limited their clinical application. It was found that collagen gels could promote the proliferation of USCs, but the underlying molecular mechanisms were unclear.
The study aims to investigate the role of the mechanically activated cation channel Piezo1 and the transcriptional coactivator YAP in the regulation of proliferation of USCs on collagen gels.
USCs were cultured on collagen gels (group COL), or plastic dishes (group NON). MTT assay, Scratch assay, EDU staining, and immunofluorescence (IF) of Ki67 were performed to evaluate the proliferation of USCs; IF of YAP was conducted to observe its nuclear localization; calcium imaging experiment was executed to evaluate the function of Piezo1; western blot was used to compare changes in protein expression of YAP, LATS1, ERK1/2, and p-ERK1/2. In addition, the regulatory effect of YAP on the proliferative capacity of USCs was confirmed by intervening YAP with its inhibitor verteporfin (VP); and the inhibitor or activator of Piezo1, GsMTx4 or Yoda1 was used to explore the effect of Piezo1 on the nuclear localization of YAP, the proliferation of USCs and the regeneration of injured bladder.
The results showed that cell proliferation was significantly enhanced in USCs in the COL group with the nuclear accumulation of YAP compared with the NON group and VP attenuated these effects. The expression and function of Piezo1 were higher in the COL group compared with the NON group. Blockage of Piezo1 by GsMTx4 decreased nuclear localization of YAP, the proliferation of USCs, and caused the failure of bladder reconstruction. Activation of Piezo1 by Yoda1 increased the nuclear expression of YAP, and the proliferation of USCs, which further improved the regeneration of the injured bladder. Finally, the ERK1/2 rather than LATS1 was revealed to participate in the Piezo1/YAP signal cascades of USCs proliferation.
Taken together, Piezo1-ERK1/2-YAP signal cascades were involved in regulating the proliferation ability of USCs in collagen gels which would be beneficial for the regeneration of the bladder.
尿液来源的干细胞（USCs）被认为是修复泌尿系统疾病的理想干细胞来源。然而，当在塑料培养皿上培养时，USCs的增殖能力显著下降，这限制了它们的临床应用。研究发现胶原蛋白凝胶可以促进USCs的增殖，但其潜在的分子机制尚不清楚。
本研究旨在探讨机械激活阳离子通道Piezo1和转录共激活因子YAP在胶原蛋白凝胶上对USCs增殖调控中的作用。
将USCs培养在胶原蛋白凝胶上（COL组）或塑料培养皿上（NON组）。采用MTT法、划痕试验、EDU染色和Ki67免疫荧光（IF）检测评估USCs的增殖；进行YAP的IF观察其核定位；进行钙成像实验评估Piezo1的功能；采用蛋白质印迹法比较YAP、LATS1、ERK1/2和p-ERK1/2蛋白表达的变化。此外，用YAP抑制剂维替泊芬（VP）干预YAP，证实YAP对USCs增殖能力的调节作用；使用Piezo1的抑制剂或激活剂GsMTx4或Yoda1，探讨Piezo1对YAP核定位、USCs增殖及膀胱损伤修复的影响。
结果显示，与NON组相比，COL组USCs的细胞增殖显著增强，YAP核内聚集增加，VP可减弱这些作用。与NON组相比，COL组Piezo1的表达和功能更高。GsMTx4阻断Piezo1可降低YAP的核定位、USCs的增殖，并导致膀胱重建失败。Yoda1激活Piezo1可增加YAP的核表达及USCs的增殖，进一步改善膀胱损伤修复。最后，发现ERK1/2而非LATS1参与了USCs增殖的Piezo1/YAP信号级联反应。
综上所述，Piezo1-ERK1/2-YAP信号级联反应参与调节胶原蛋白凝胶上USCs的增殖能力，这将有利于膀胱的修复。
