The Garlic Compound Z-Ajoene, S-Thiolates COX2 and STAT3 and Dampens the Inflammatory Response in RAW264.7 Macrophages
The Garlic Compound Z-Ajoene, S-Thiolates COX2 and STAT3 and Dampens the Inflammatory Response in RAW264.7 Macrophages
复制标题
DOI：
10.1002/mnfr.202000854
复制
发表时间：
2020-12-18
影响因子：
5.2
通讯作者：
Kaschula, Catherine H.
中科院分区：
文献类型：
--
作者：
Hitchcock, Jessica K.;Mkwanazi, Nonkululeko;Kaschula, Catherine H.
范围大蒜（Allium sativum）几个世纪以来一直被用作预防和治疗药物，以控制炎症相关的病理。为了研究潜在的机制，在大蒜化合物大蒜素和Z-阿焦烯（ZA）的存在下，使用暴露于低剂量脂多糖（LPS）的RAW 264.7小鼠巨噬细胞建立体外炎症模型，模拟常规的大蒜消耗。并上调抗炎细胞因子IL 10的表达。选择分泌的炎症介质的蛋白质阵列证实，Z-阿焦烯对LPS诱导的炎症细胞因子和趋化因子具有显著的下调作用。这些蛋白质中的许多是转录因子信号转导子和转录激活子3（STAT 3）的已知靶点;事实上，发现Z-阿焦烯或其类似物丹酰-阿焦烯减少STAT 3的磷酸化和核转位，并通过Cys 108、Cys 367和Cys 687处的S-巯基化共价修饰蛋白质。Z-ajoene对COX 2的抑制作用呈剂量依赖性，且呈非竞争性，可能与Cys 9和Cys 299的巯基化有关。结论Z-ajoene对炎症调节因子STAT 3和COX 2的靶向修饰作用可能与大蒜的健康效应有关。
Scope Garlic (Allium sativum) has been used for centuries as a prophylactic and therapeutic medicinal agent to control inflammation-associated pathologies. To investigate the underlying mechanisms, an in vitro inflammatory model is established using RAW264.7 murine macrophages exposed to low-doses of lipopolysaccharide (LPS) in the presence of garlic compounds allicin and Z-ajoene (ZA), mimicking regular garlic consumption.Methods and Results Both allicin and Z-ajoene dampen both transcript and protein expression of the pro-inflammatory cytokines IL1 beta, IL6, and IL12 beta, and upregulate the expression of the anti-inflammatory cytokine IL10. Protein arrays of selected secreted inflammatory mediators confirm that Z-ajoene has a pronounced down-regulatory effect on LPS-induced inflammatory cytokines and chemokines. Many of these proteins are known targets of the transcription factor signal transducer and activator of transcription 3 (STAT3); and indeed, Z-ajoene or its analogue dansyl-ajoene is found to decrease phosphorylation and nuclear translocation of STAT3, and to covalently modify the protein by S-thiolation at Cys108, Cys367, and Cys687. Z-Ajoene dose-dependently and non-competitively inhibit the activity of cyclooxygenase 2 (COX2), possibly attributed to S-thiolation at Cys9 and Cys299.Conclusion The characterization of Z-ajoene's activity of targeting and covalently modifying STAT3 and COX2, both important regulators of inflammation, may contribute to the health benefits of regular dietary garlic consumption.