Abstract：
Daruqi is a Traditional Mongolian medicine with anti-inflammatory, anti-bacterial, and immune-regulatory effects. However, the mechanisms of its activity were unclear. In the present study, we confirmed the anti-inflammation effect of Daruqi on inflammation induced by LPS using animal models. Then, THP-1 cells treated with LPS was used as a positive control to explore the effective component of Daruqi on inflammation. We identified that Oxymatrine was the essential effector of Daruqi. Furthermore, the mechanism of Oxymatrine on inflammation was verified through proteomics analyses and validation assays. Our results demonstrated that Oxymatrine significantly reduced the levels of inflammatory cytokine, including IL-8, IL-1α, and IL-1β, in LPS induced THP-1 cells. Based on tandem mass tag -labeled quantitative proteomics, 428 differentially expressed proteins were screened, involved in TNF signaling pathway, Ferroptosis, IL-17 signaling pathway, etc. Among these differential expressed proteins (DEPs), 23 proteins were verified with parallel reaction monitoring analysis. The results showed that LPS treatment potentiated the protein level of PLEK, ACSL5 and CYBB, which could be reversed by Oxymatrine. By contrast, the protein expression of SPRYD4 and EMR2 was suppressed after LPS treatment, which could be rescued by Oxymatrine. In summary, Oxymatrine has excellent protective effects in LPS induced THP-1 cells. The five proteins, including PLEK, ACSL5, CYBB, SPRYD4 and EMR2, might serve as the targets of Oxymatrine, and as candidates regulating inflammation in future therapies.
摘要：
达鲁奇是一种具有抗炎作用的传统蒙药，抗菌,和免疫调节作用。然而,其活动机制尚不清楚。在本研究中,我们用动物模型证实了达鲁奇对LPS诱导的炎症的抗炎作用。然后，以LPS处理的THP-1细胞作为阳性对照,探讨达鲁芪的有效成分对炎症的影响。我们确定氧化苦参碱是达鲁奇的重要效应物。此外，通过蛋白质组学分析和验证试验验证了氧化苦参碱对炎症的作用机制.我们的结果表明，氧化苦参碱显着降低炎症细胞因子的水平，包括IL-8，IL-1α，和IL-1β，在LPS诱导的THP-1细胞中。基于串联质量标签标记的定量蛋白质组学，筛选出428个差异表达蛋白,参与TNF信号通路，Ferroptosis,IL-17信号通路,等。在这些差异表达蛋白（DEP）中，用平行反应监测分析验证23种蛋白质。结果表明，LPS处理增强了PLEK的蛋白质水平，ACSL5和CYBB，氧化苦参碱可以逆转。相比之下,LPS处理后，SPRYD4和EMR2的蛋白表达受到抑制，可以用氧化苦参碱来拯救.总之,氧化苦参碱对LPS诱导的THP-1细胞具有良好的保护作用。五种蛋白质，包括PLEK,ACSL5,CYBB,SPRYD4和EMR2可能作为氧化苦参碱的靶标，并作为未来治疗中调节炎症的候选人。
