Investigation of the Anti-Inflammatory Activity of Fusaproliferin Analogues Guided by Transcriptome Analysis.
Investigation of the Anti-Inflammatory Activity of Fusaproliferin Analogues Guided by Transcriptome Analysis.
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DOI：
10.3389/fphar.2022.881182
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发表时间：
2022
影响因子：
5.6
通讯作者：
Guo, Da-Le
中科院分区：
文献类型：
--
作者：
Kuang, Qi-Xuan;Lei, Li-Rong;Li, Qing-Zhou;Peng, Wan;Wang, Yu-Mei;Dai, Yi-Fei;Wang, Dong;Gu, Yu-Cheng;Deng, Yun;Guo, Da-Le
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背景资料：过度的炎症会导致严重的组织损伤以及严重的急性或慢性疾病，广泛的研究集中在从天然产品中寻找具有安全性和有效性的新的抗炎化合物。作为治疗炎症相关疾病的有前景的治疗实体，融合蛋白及其类似物引起了人们极大的兴趣。然而，潜在的抗炎机制仍然知之甚少，值得进一步研究。 研究方法：为了评估融合蛋白（1）及其类似物（2 - 4）在体外和体内的抗炎活性，使用脂多糖（LPS）诱导的RAW 264.7巨噬细胞和斑马鱼胚胎。然后，应用转录组分析来指导随后的相关信号通路中的关键蛋白质的蛋白质印迹分析。表面等离子体共振分析（SPR）结合分子对接分析最终用于评估1 - 4与TLR4之间的亲和力相互作用，并提供相关信号通路下调的可能解释。 结果如下：1 - 4显著减弱LPS诱导的RAW 264.7巨噬细胞中炎性信使的产生，包括一氧化氮（NO）、活性氧（ROS）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1 β（IL-1 β），以及一氧化氮合酶（iNOS）和环氧合酶-2（考克斯-2）。基于RNA-seq的转录组分析表明化合物1逆转LPS刺激的能力和核因子κ-B（NF-κ B）和促分裂原活化蛋白激酶（MAPK）信号传导途径有助于抗炎过程。蛋白水平的实验验证表明，1能抑制NF-κ B激酶抑制剂（IKK）的活化、NF-κ B抑制剂（I κ B）的降解、NF-κ B的磷酸化，减少NF-κ B的核转位。1还降低MAPK的磷酸化，包括p38、细胞外调节蛋白激酶（ERK）和c-Jun N-末端激酶（JNK）。SPR分析和分子对接结果表明，1 - 4对TLR4蛋白具有亲和力，KD值为23.5 - 29.3 μ M。 结论：Fusaproliferin及其类似物是治疗炎症相关疾病的理想化合物。
Background: Excessive inflammation results in severe tissue damage as well as serious acute or chronic disorders, and extensive research has focused on finding new anti-inflammatory hit compounds with safety and efficacy profiles from natural products. As promising therapeutic entities for the treatment of inflammation-related diseases, fusaproliferin and its analogs have attracted great interest. However, the underlying anti-inflammatory mechanism is still poorly understood and deserves to be further investigated. Methods: For the estimation of the anti-inflammatory activity of fusaproliferin (1) and its analogs (2-4) in vitro and in vivo, lipopolysaccharide (LPS)-induced RAW264.7 macrophages and zebrafish embryos were employed. Then, transcriptome analysis was applied to guide subsequent western blot analysis of critical proteins in related signaling pathways. Surface plasmon resonance assays (SPR) combined with molecular docking analyses were finally applied to evaluate the affinity interactions between 1-4 and TLR4 and provide a possible interpretation of the downregulation of related signaling pathways. Results: 1-4 significantly attenuated the production of inflammatory messengers, including nitric oxide (NO), reactive oxygen species (ROS), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), as well as nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in LPS-induced RAW264.7 macrophages. Transcriptome analyses based on RNA-seq indicated the ability of compound 1 to reverse LPS stimulation and the nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPKs) signaling pathways contribute to the anti-inflammatory process. Experimental verification at the protein level revealed that 1 can inhibit the activation of inhibitor of NF-κB kinase (IKK), degradation of inhibitor of NF-κB (IκB), and phosphorylation of NF-κB and reduce nuclear translocation of NF-κB. 1 also decreased the phosphorylation of MAPKs, including p38, extracellular regulated protein kinases (ERK), and c-Jun N-terminal kinase (JNK). SPR assays and molecular docking results indicated that 1-4 exhibited affinity for the TLR4 protein with KD values of 23.5–29.3 μM. Conclusion: Fusaproliferin and its analogs can be hit compounds for the treatment of inflammation-associated diseases.
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4.7
作者：
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