The anti-alcoholism drug disulfiram effectively ameliorates ulcerative colitis through suppressing oxidative stresses-associated pyroptotic cell death and cellular inflammation in colonic cells
The anti-alcoholism drug disulfiram effectively ameliorates ulcerative colitis through suppressing oxidative stresses-associated pyroptotic cell death and cellular inflammation in colonic cells
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抗酒精药物双硫仑通过抑制结肠细胞中与氧化应激相关的焦亡细胞死亡和细胞炎症来有效改善溃疡性结肠炎。
DOI：
10.1016/j.intimp.2022.109117
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发表时间：
2022-08-12
影响因子：
5.6
通讯作者：
Zhu, Yuekun
中科院分区：
文献类型：
--
作者：
Chi, Fengxu;Zhang, Guangquan;Zhu, Yuekun
背景：氧化应激、细胞焦亡和炎症被认为是溃疡性结肠炎（UC）发展的重要致病因素，而传统的解酒药物双硫仑（DSF）最近被报道对上述所有细胞功能发挥调节作用，这使得DSF成为UC治疗的理想治疗剂，但这一问题尚未得到充分研究。方法：C57BL/6J小鼠和脂多糖诱导的右旋糖酐硫酸钠（DSS）动物模型分别建立了LPS诱导的UC结肠细胞系（HT-29和Caco-2）细胞模型。通过ELISA测定细胞因子分泌。通过MTT测定和EdU测定评估细胞活力和增殖。采用实时qPCR、Western Blot、免疫荧光染色测定和免疫组织化学（IHC）来评估基因表达。采用 Pearson 相关分析对临床组织中基因的相关性进行分析。结果：DSF 在体外和体内 UC 模型中抑制氧化应激、焦亡细胞死亡和细胞炎症，并且通过 N-乙酰基-L-半胱氨酸 (NAC) 消除活性氧 (ROS)，挽救了 LPS 处理的结肠细胞（HT-29 和 Caco-2）的细胞活力。进一步的实验表明，糖原合成酶激酶 3 beta (GSK-3 beta)/Nrf2/NLRP3 信号级联在此过程中发挥了关键作用。从机制上讲，DSF 下调 GSK-3 beta 和 NLRP3，而上调 LPS 处理的结肠细胞中的 Nrf2。此外，DSF 对 Nrf2 和 NLRP3 的调节作用可以通过上调 GSK-3 beta 来消除。此外，GSK-3β的上调消除了DSF对LPS处理的结肠细胞的保护作用。结论：总而言之，本研究的数据表明，DSF通过调节GSK-3β/Nrf2/NLRP3途径抑制氧化损伤相关的焦亡细胞死亡和炎症，从而抑制LPS诱导的UC发展。
Background: Oxidative stress, cell pyroptosis and inflammation are considered as important pathogenic factors for ulcerative colitis (UC) development, and the traditional anti-alcoholism drug disulfiram (DSF) has recently been reported to exert its regulating effects on all the above cellular functions, which makes DSF as ideal therapeutic agent for UC treatment, but this issue has not been fully studied.Methods: Dextran sulfate sodium (DSS)-induced animal models in C57BL/6J mice and lipopolysaccharide (LPS)-induced cellular models in colonic cell lines (HT-29 and Caco-2) for UC were respectively established. Cytokine secretion was determined by ELISA. Cell viability and proliferation were evaluated by MTT assay and EdU assay. Real-Time qPCR, Western Blot, immunofluorescent staining assay and immunohistochemistry (IHC) were employed to evaluate gene expressions. The correlations of the genes in the clinical tissues were analyzed by using the Pearson Correlation analysis.Results: DSF restrained oxidative stress, pyroptotic cell death and cellular inflammation in UC models in vitro and in vivo, and elimination of Reactive Oxygen Species (ROS) by N-acetyl-L-cysteine (NAC) rescued cell viability in LPS-treated colonic cells (HT-29 and Caco-2). Further experiments suggested that a glycogen synthase kinase-3 beta (GSK-3 beta)/Nrf2/NLRP3 signaling cascade played critical role in this process. Mechanistically, DSF downregulated GSK-3 beta and NLRP3, whereas upregulated Nrf2 in LPS-treated colonic cells. Also, the regulating effects of DSF on Nrf2 and NLRP3 were abrogated by upregulating GSK-3 beta. Moreover, upregulation of GSK-3 beta abolished the protective effects of DSF on LPS-treated colonic cells.Conclusions: Taken together, data of this study indicated that DSF restrained oxidative damages-related pyroptotic cell death and inflammation via regulating the GSK-3 beta/Nrf2/NLRP3 pathway, leading to the suppression of LPS-induced UC development.