克罗恩病（CD）的长期预后仍然不理想。因此，我们评估了胸腺五肽（TP5）对2,4,6-三硝基苯磺酸（TNBS）诱导结肠炎小鼠模型的治疗效果，并分析了其对中性粒细胞外细胞陷阱（NETs）的影响。通过免疫染色和酶联免疫吸附法评估NET标志物，包括髓过氧化物酶（MPO）、中性粒细胞弹力蛋白酶（NE）、柠檬酰化组蛋白H3（CitH3）、肽酰精氨酸脱互异化酶IV（PAD4）和双链DNA（dsDNA）。通过体外评估NET形成。使用新清蛋白3，特定NET激动剂，来逆转TP5对TNBS诱导结肠炎的影响。用RNA-seq研究TP5的作用机制.TP5改善了TNBS组体重下降（P < 0.001），疾病活动指数（DAI）（P = 0.05），结肠萎缩（P = 0.04），以及肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6和中性粒细胞水平的升高。TNBS组MPO、NE、CitH3、PAD4、dsDNA和MPO-DNA水平增加（P < 0.001），在TP5治疗后减少（P = 0.01, P < 0.001, P < 0.001, P < 0.001, P = 0.02 和 P = 0.02）。组织CitH3水平与DAI和TNF-α水平呈正相关（P < 0.05）。此外，在体外刺激凋亡素12-肉豆蔻酸13-乙酯诱导的NET形成在对照组、TNBS+生理盐水组和TNBS+TP5组中分别增加了1.8倍、2.8倍和2.3倍。新清蛋白3显著逆转了TP5的效果。RNA-seq揭示了TP5效果的潜在途径.TP5通过抑制NETs有效缓解了实验CD模型的结肠炎。版权所有 © 2023 Elsevier B.V. 保留所有权利。

The long-term prognosis of Crohn's disease (CD) remains unsatisfactory. Therefore, we assessed the therapeutic effect of thymopentin (TP5) in a mouse model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis, which mimics CD, and analyzed its impact on neutrophil extracellular traps (NETs).NET markers, including myeloperoxidase (MPO), neutrophil elastase (NE), citrullinated histone H3 (CitH3), peptidyl arginine deiminase IV (PAD4), and double-stranded DNA (dsDNA) were assessed by immunostaining and enzyme-linked immunosorbent assay. NET formation was evaluated in vitro. Neoseptin 3, a specific NET agonist, was used to reverse the effect of TP5 on TNBS-induced colitis. The action mechanism of TP5 was investigated using RNA-seq.TP5 ameliorated weight loss (P < 0.001), disease activity index (DAI) (P = 0.05), colon shrinkage (P = 0.04), and elevated levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-6, and neutrophils in the TNBS group. The TNBS group exhibited increased MPO, NE, CitH3, PAD4, dsDNA and MPO-DNA levels (all P < 0.001), which decreased after TP5 administration (P = 0.01, P < 0.001, P < 0.001, P < 0.001, P = 0.02, and P = 0.02 respectively). Tissue CitH3 levels were positively correlated with DAI and TNF-α levels (P < 0.05). Furthermore, phorbol 12-myristate 13-acetate-stimulated NET formation increased by 1.8-, 2.8-, and 2.3-fold in vitro in the control, TNBS + saline, and TNBS + TP5 groups, respectively. Neoseptin 3 significantly reversed the effect of TP5. RNA-seq revealed potential pathways underlying the effect of TP5.TP5 effectively ameliorated colitis by suppressing NETs in the experimental CD model.Copyright © 2023 Elsevier B.V. All rights reserved.