Division of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, Hubei Province 430022, China.
Division of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, Hubei Province 430022, China.
Life Sci. 2021 Jun 15;275:119254. doi: 10.1016/j.lfs.2021.119254. Epub 2021 Feb 24.
Defective tight junctions (TJs) can induce intestinal epithelial dysfunction, which participates in various diseases such as irritable bowel syndrome. However, the mechanisms of TJ defects remain unclear. Our study revealed the role of Piezo1 in regulating intestinal epithelial function and TJs.
The human colonic adenocarcinoma cell line Caco-2 were cultured on Transwell plate to form an epithelial barrier in vitro, and Piezo1 expression was manipulated using a lentivirus vector. Epithelial function was evaluated by measuring transepithelial electronic resistance (TEER) and 4-kDa FITC-dextran (FD4) transmission. TJ proteins (claudin-1, occludin, ZO-1) were evaluated by RT-PCR, western blot, and immunostaining analysis. Potential signal pathways, including the ROCK and Erk pathways, were detected. Moreover, to explore the regulatory effect of Piezo1 activity on epithelial function, inhibitors (ruthenium red, GsMTx4) and an agonist (Yoda1) were introduced both ex vivo and in vitro.
Alteration of Piezo1 expression altered epithelial function and the expression of the tight junction protein claudin-1. Piezo1 expression regulated phosphorylated ROCK1/2 expression, whereas interference on ROCK1/2 prevented the regulation of claudin-1 by Piezo1. In both Caco-2 monolayer and mouse colon epithelium, Piezo1 activity directly modulated epithelial function and permeability.
Piezo1 negatively regulates epithelial barrier function by affecting the expression of claudin-1. Such regulation may be achieved partially via the ROCK1/2 pathway. Moreover, activating Piezo1 can induce epithelial dysfunction.
有缺陷的紧密连接（TJ）可诱导肠道上皮功能障碍，参与多种疾病，如肠易激综合征。然而，TJ 缺陷的机制尚不清楚。我们的研究揭示了 Piezo1 在调节肠道上皮功能和 TJ 中的作用。
体外培养人结肠腺癌细胞系 Caco-2 在 Transwell 板上形成上皮屏障，使用慢病毒载体操纵 Piezo1 表达。通过测量跨上皮电阻（TEER）和 4-kDa FITC-右旋糖酐（FD4）传递来评估上皮功能。通过 RT-PCR、western blot 和免疫染色分析评估 TJ 蛋白（claudin-1、occludin、ZO-1）。检测了潜在的信号通路，包括 ROCK 和 Erk 通路。此外，为了探讨 Piezo1 活性对上皮功能的调节作用，在体外和体内引入了抑制剂（钌红、GsMTx4）和激动剂（Yoda1）。
Piezo1 表达的改变改变了上皮功能和紧密连接蛋白 claudin-1 的表达。Piezo1 表达调节磷酸化 ROCK1/2 表达，而 ROCK1/2 的干扰阻止了 Piezo1 对 claudin-1 的调节。在 Caco-2 单层和小鼠结肠上皮中，Piezo1 活性直接调节上皮功能和通透性。
Piezo1 通过影响 claudin-1 的表达来负调控上皮屏障功能。这种调节可能部分通过 ROCK1/2 通路实现。此外，激活 Piezo1 可诱导上皮功能障碍。
