Cao Dongxing, Qian Keyu, Yang Nailin, Xu Gang, Wang Xiaohui, Zhu Mingming, Wang Yangyang, Li Han, Shen Jun, Zhang Ye, Cui Zhe
Department of General Surgery, Baoshan Branch, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200444, China; Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
Laboratory of Medicine, Baoshan Branch, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200444, China.
Int Immunopharmacol. 2023 Nov;124(Pt A):110898. doi: 10.1016/j.intimp.2023.110898. Epub 2023 Sep 9.
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.
克罗恩病（CD）的长期预后仍不尽人意。因此，我们评估了胸腺五肽（TP5）在2,4,6-三硝基苯磺酸（TNBS）诱导的结肠炎小鼠模型中的治疗效果，该模型可模拟CD，并分析其对中性粒细胞胞外诱捕网（NETs）的影响。
通过免疫染色和酶联免疫吸附测定评估包括髓过氧化物酶（MPO）、中性粒细胞弹性蛋白酶（NE）、瓜氨酸化组蛋白H3（CitH3）、肽基精氨酸脱亚氨酶IV（PAD4）和双链DNA（dsDNA）在内的NET标志物。在体外评估NET的形成。使用特异性NET激动剂Neoseptin 3来逆转TP5对TNBS诱导的结肠炎的作用。使用RNA测序研究TP5的作用机制。
TP5改善了体重减轻（P < 0.001）、疾病活动指数（DAI）（P = 0.05）、结肠收缩（P = 0.04），并降低了TNBS组中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6和中性粒细胞的水平升高。TNBS组的MPO、NE、CitH3、PAD4、dsDNA和MPO-DNA水平升高（均P < 0.001），给予TP5后降低（分别为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，佛波酯12-肉豆蔻酸酯13-乙酸酯刺激的NET形成在对照组、TNBS + 盐水组和TNBS + TP5组中分别增加了1.8倍、2.8倍和2.3倍。Neoseptin 3显著逆转了TP5的作用。RNA测序揭示了TP5作用的潜在途径。
在实验性CD模型中，TP5通过抑制NETs有效改善了结肠炎。
