肠道干细胞（ISC）在维持肠道组织的平衡和再生潜力方面发挥着至关重要的作用，从而确保组织稳态并促进损伤后有效的组织再生。事实证明，靶向 Toll 样受体 (TLR) 有助于预防辐射引起的肠道损伤。在本研究中，我们利用IR建立了肠道损伤模型，并在体内和体外评估了CL429对ISC再生的影响。辐射暴露后，接受 CL429 治疗的小鼠的存活率显着提高（治疗组的存活率为 100%，而对照组的存活率为 54.54%）。 CL429在抑制辐射引起的肠道损伤和促进ISC增殖和再生方面也表现出显着的功效。此外，CL429 还可以保护肠道类器官免受红外线诱导的损伤。从机制上讲，RNA 测序和蛋白质印迹分析揭示了 CL429 对 Wnt 和 Hippo 信号通路的激活。具体来说，我们观察到 YAP1（Hippo 通路中的关键转录因子）在 CL429 刺激后显着上调。此外，YAP1的敲低显着减弱了CL429对肠道类器官的辐射防护作用，表明CL429介导的肠道辐射防护依赖于YAP1。此外，我们使用TLR2敲除小鼠研究了TLR2和YAP1之间的关系，结果表明TLR2敲除消除了YAP1上CL429的激活。综上所述，我们的研究提供了证据支持 CL429 通过激活 TLR2-YAP1 促进 ISC 再生的作用。进一步研究 TLR 与其他信号通路之间的相互作用可能会增强我们对损伤后 ISC 再生的理解。版权所有 © 2024。由 Elsevier B.V. 出版。

Intestinal stem cells (ISCs) play a crucial role in maintaining the equilibrium and regenerative potential of intestinal tissue, thereby ensuring tissue homeostasis and promoting effective tissue regeneration following injury. It has been proven that targeting Toll-like receptors (TLRs) can help prevent radiation-induced damage to the intestine. In this study, we established an intestinal injury model using IR and evaluated the effects of CL429 on ISC regeneration both in vivo and in vitro. Following radiation exposure, mice treated with CL429 showed a significant increase in survival rates (100% survival in the treated group compared to 54.54% in the control group). CL429 also showed remarkable efficacy in inhibiting radiation-induced intestinal damage and promoting ISC proliferation and regeneration. In addition, CL429 protected intestinal organoids against IR-induced injury. Mechanistically, RNA sequencing and Western blot analysis revealed the activation of the Wnt and Hippo signaling pathways by CL429. Specifically, we observed a significant upregulation of YAP1, a key transcription factor in the Hippo pathway, upon CL429 stimulation. Furthermore, knockdown of YAP1 significantly attenuated the radioprotective effect of CL429 on intestinal organoids, indicating that CL429-mediated intestinal radioprotection is dependent on YAP1. In addition, we investigated the relationship between TLR2 and YAP1 using TLR2 knockout mice, and our results showed that TLR2 knockout abolished the activation of CL429 on YAP1. Taken together, our study provides evidence supporting the role of CL429 in promoting ISC regeneration through activation of TLR2-YAP1. And further investigation of the interaction between TLRs and other signaling pathways may enhance our understanding of ISC regeneration after injury.Copyright © 2024. Published by Elsevier B.V.