抑制第 3 组先天淋巴细胞 (ILC3) 中的死亡受体 3 (DR3) 信号通路为促进溃疡性结肠炎 (UC) 患者的粘膜修复提供了一种有前景的方法。芍药甙是芍药的主要成分，已被证明能够恢复UC小鼠的屏障功能，但其确切机制仍不清楚。在本研究中，我们旨在深入探讨芍药苷是否可以通过抑制ILC3中的DR3信号传导来促进慢性结肠炎的肠粘膜修复。 C57BL/6 小鼠随机分为 7 个不同组，即对照组、2% 右旋糖酐硫酸钠 (DSS) 组、芍药苷组（25、50 和 100 mg/kg）、抗肿瘤坏死组因子样配体1A（抗TL1A）抗体组和IgG组。我们分别使用Western blot和流式细胞术检测小鼠结肠中DR3信号通路蛋白的表达和ILC3的比例。同时，使用 DR3 过表达的 MNK-3 细胞和 2% DSS 诱导的 Rag1-/- 小鼠进行验证。结果表明，芍药苷可缓解DSS引起的慢性结肠炎并修复肠粘膜屏障。同时，芍药苷抑制ILC3中的DR3信号通路并调节细胞因子（白细胞介素17A、粒细胞-巨噬细胞集落刺激因子和白细胞介素22）的含量。或者，芍药苷直接抑制 ILC3 中的 DR3 信号通路，以独立于适应性免疫系统修复粘膜损伤。我们还证实，芍药苷条件培养基 (CM) 通过共培养恢复了 Caco-2 细胞中紧密连接的表达。总之，芍药苷通过ILC3依赖性增强肠道屏障来改善慢性结肠炎，其机制与抑制DR3信号通路有关。© 2024 The Authors。

Inhibiting the death receptor 3 (DR3) signaling pathway in group 3 innate lymphoid cells (ILC3s) presents a promising approach for promoting mucosal repair in individuals with ulcerative colitis (UC). Paeoniflorin, a prominent component of Paeonia lactiflora Pall., has demonstrated the ability to restore barrier function in UC mice, but the precise mechanism remains unclear. In this study, we aimed to delve into whether paeoniflorin may promote intestinal mucosal repair in chronic colitis by inhibiting DR3 signaling in ILC3s. C57BL/6 mice were subjected to random allocation into 7 distinct groups, namely the control group, the 2 % dextran sodium sulfate (DSS) group, the paeoniflorin groups (25, 50, and 100 mg/kg), the anti-tumor necrosis factor-like ligand 1A (anti-TL1A) antibody group, and the IgG group. We detected the expression of DR3 signaling pathway proteins and the proportion of ILC3s in the mouse colon using Western blot and flow cytometry, respectively. Meanwhile, DR3-overexpressing MNK-3 cells and 2 % DSS-induced Rag1-/- mice were used for verification. The results showed that paeoniflorin alleviated DSS-induced chronic colitis and repaired the intestinal mucosal barrier. Simultaneously, paeoniflorin inhibited the DR3 signaling pathway in ILC3s and regulated the content of cytokines (Interleukin-17A, Granulocyte-macrophage colony stimulating factor, and Interleukin-22). Alternatively, paeoniflorin directly inhibited the DR3 signaling pathway in ILC3s to repair mucosal damage independently of the adaptive immune system. We additionally confirmed that paeoniflorin-conditioned medium (CM) restored the expression of tight junctions in Caco-2 cells via coculture. In conclusion, paeoniflorin ameliorates chronic colitis by enhancing the intestinal barrier in an ILC3-dependent manner, and its mechanism is associated with the inhibition of the DR3 signaling pathway.© 2024 The Authors.