Demethylzeylasteral alleviates inflammation and colitis via dual suppression of NF-κB and STAT3/5 by targeting IKKα/β and JAK2.

IF 4.8 2区医学 Q2 IMMUNOLOGY

International immunopharmacology Pub Date : 2024-12-05 Epub Date: 2024-09-27 DOI:10.1016/j.intimp.2024.113260

Tian Wen, Ting Liu, Hongqing Chen, Qi Liu, Xiaofei Shen, Qiongying Hu

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引用次数: 0

Abstract

Background: Ulcerative colitis (UC) is a common inflammatory bowel disease and a risk factor of colorectal cancer. Demethylzeylasteral (DZT), a bioactive component mainly isolated from Tripterygium wilfordii, has been shown to inhibit inflammation and cancer. However, its anti-UC function and molecular mechanisms have not been well characterized. This study aims to explore the therapeutic effect and functional targets of demethylzeylasteral against UC.

Methods: RT-qPCR, Western blot and ELISA were used to detect the generation of pro-inflammatory cytokines and chemokines in murine macrophage cells. Luciferase reporter gene, Western blot, pull-down, CETSA, DARTS, and virtual docking were employed to detect the anti-inflammatory targets and molecular mechanisms of demethylzeylasteral. The anti-inflammatory and anti-colitis effects of demethylzeylasteral were further determined in DSS-challenged mice.

Results: In vitro, demethylzeylasteral inhibited NO and PGE₂ production by suppressing the mRNA and protein expression of iNOS and COX-2, and suppressed the mRNA expression of TNF-α, IL-1β, IL-6, MCP-1, CXCL9, and CXCL10 in RAW264.7 macrophages stimulated by LPS/IFNγ. Furthermore, demethylzeylasteral was not only capable of inhibiting IKKα/β-NF-κB activation, but also able to block JAKs-STAT3/5 activation in LPS/INFγ-incubated RAW264.7 cells or DSS-exposed colon tissues of mice. Mechanistically, demethylzeylasteral was found to directly bind to IKKα/β and JAK2 kinases, leading to inactivation of pro-inflammatory signaling cascades and reduced generation of cytokines and chemokines. In vivo, oral administration of demethylzeylasteral significantly attenuated DSS-induced colitis, which was mainly manifested as mitigated symptoms of colitis, colonic mucosal barrier damage, and colonic inflammation.

Conclusion: We demonstrated that demethylzeylasteral alleviated UC pathology by blocking NF-κB and STAT3/5 pathways via targeting IKKα/β and JAK2 kinases, raising the possibility that demethylzeylasteral could act as a candidate for the treatment of UC.

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去甲斑蝥素通过靶向 IKKα/β 和 JAK2 双重抑制 NF-κB 和 STAT3/5，从而缓解炎症和结肠炎。

背景：溃疡性结肠炎（UC溃疡性结肠炎（UC）是一种常见的炎症性肠病，也是结直肠癌的危险因素之一。去甲斑蝥素（Demethylzeylasteral，DZT）是一种生物活性成分，主要从三尖杉（Tripterygium wilfordii）中分离出来，已被证明具有抑制炎症和癌症的作用。然而，其抗 UC 功能和分子机制尚未得到很好的表征。本研究旨在探索去甲斑蝥素对 UC 的治疗效果和功能靶点：方法：采用 RT-qPCR、Western blot 和 ELISA 检测小鼠巨噬细胞中促炎细胞因子和趋化因子的生成。采用荧光素酶报告基因、Western blot、pull-down、CETSA、DARTS和虚拟对接等方法检测去甲斑蝥素的抗炎靶点和分子机制。结果表明：在体外，去甲斑蝥素具有抗炎和抗胆囊炎的作用：结果：在体外，去甲泽泻通过抑制 iNOS 和 COX-2 的 mRNA 和蛋白表达来抑制 NO 和 PGE2 的产生，并抑制 LPS/IFNγ 刺激的 RAW264.7 巨噬细胞中 TNF-α、IL-1β、IL-6、MCP-1、CXCL9 和 CXCL10 的 mRNA 表达。此外，去甲斑蝥素不仅能抑制 IKKα/β-NF-κB 的活化，还能阻断 LPS/INFγ 培养的 RAW264.7 细胞或小鼠暴露于 DSS 的结肠组织中 JAKs-STAT3/5 的活化。从机理上讲，去甲斑蝥素可直接与 IKKα/β 和 JAK2 激酶结合，导致促炎信号级联失活，并减少细胞因子和趋化因子的生成。在体内，口服去甲斑蝥素可显著减轻DSS诱导的结肠炎，主要表现为减轻结肠炎症状、结肠粘膜屏障损伤和结肠炎症：结论：我们证明了去甲斑蝥素通过靶向IKKα/β和JAK2激酶阻断NF-κB和STAT3/5通路，从而减轻了UC的病理变化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.