PDF(Pubmed)
Abstract：
Chrysanthemi indic Flos (CIF) has been commonly consumed for the treatment of inflammation and related skin diseases. However, the potential bioactive components responsible for its anti-inflammatory and sensitive skin (SS) improvement activities, and the correlated mechanisms of action still remain unknown. In this work, it was firstly found that the CIF extract (CIFE) displayed arrestive free radical scavenging activity on DPPH and ABTS radicals, with no significant difference with positive control Trolox (p > 0.05). Then, compared to the negative group, CIFE markedly decreased the productions of the pro-inflammatory cytokines (IL-1β, IL-6, PEG2, TNF-α, IFN-γ, NO) in LPS induced RAW264.7 cells in a dose-dependent manner (p < 0.01). Besides, CIFE strongly inhibited the COX-2 and hyaluronidase (HAase) with the IC50 values of 1.06 ± 0.01 μg/mL and 12.22 ± 0.39 μg/mL, indicating higher inhibitory effect than positive control of aspirin of 6.33 ± 0.05 μg/mL (p < 0.01), and comparable inhibitory effect with indometacin of 0.60 ± 0.03 μg/mL, and ascorbic acid of 11.03 ± 0.41 μg/mL (p > 0.05), respectively. Furthermore, kinetic assays with Lineweaver-Burk plot (Michaelis Menten equation) suggested that CIFE reversibly inhibited the COX-2 and HAase, with a mixed characteristics of competitive and non-competitive inhibition. Thereafter, multi-target affinity ultrafiltration liquid chromatography-mass spectrometry (UF-LC/MS) method was employed to fast fish out the potential COX-2 and HAase in CIFE. Herein, 13 components showed various affinity binding degrees to the COX-2 and HAase, while those components with relative binding affinity (RBA) value higher than 3.0, such as linarin and chlorogenic acid isomers, were deemed to be the most bioactive components for the anti-inflammatory and SS improvement activities of CIFE. Finally, the interaction mechanism, including binding energy, inhibition constant, docking sites, and the key amino acids involved in hydrogen bonds between the potential ligands and COX-2/HAase were simulated and confirmed with the molecule docking analysis. In summary, this study showcased the prominent anti-inflammatory and SS improvement activities of CIF, which would provide further insights on this functional medicinal plant to be a natural anti-SS remedy.
摘要：
菊花（CIF）通常用于治疗炎症和相关皮肤病。然而,负责其抗炎和敏感皮肤（SS）改善活性的潜在生物活性成分，相关的作用机制仍然未知。在这项工作中,首次发现CIF提取物（CIFE）对DPPH和ABTS自由基具有明显的自由基清除活性，与阳性对照Trolox无显著差异(p>0.05)。然后，与阴性组相比，CIFE显着降低了促炎细胞因子（IL-1β，IL-6,PEG2,TNF-α,IFN-γ，NO)在LPS诱导RAW264.7细胞中呈剂量依赖性(p<0.01)。此外,CIFE强烈抑制COX-2和透明质酸酶（HAase），IC50值为1.06±0.01μg/mL和12.22±0.39μg/mL，表明阿司匹林6.33±0.05μg/mL的抑制作用高于阳性对照（p<0.01），与0.60±0.03μg/mL的吲哚美辛的抑制作用相当，抗坏血酸为11.03±0.41μg/mL(p>0.05)，分别。此外，用Lineweaver-Burk图（MichaelisMenten方程）进行的动力学测定表明，CIFE可逆地抑制COX-2和HAase，具有竞争性和非竞争性抑制的混合特征。此后,采用多靶标亲和超滤液相色谱-质谱（UF-LC/MS）方法快速提取了CIFE中潜在的COX-2和HAase。在这里，13个组分对COX-2和HAase表现出不同的亲和力结合程度，而相对结合亲和力(RBA)值高于3.0的组分,如linarin和绿原酸异构体,被认为是CIFE抗炎和SS改善活性的最具生物活性的成分。最后,相互作用机制，包括结合能，抑制常数，对接点，通过分子对接分析，模拟并确认了参与潜在配体与COX-2/HAase之间氢键的关键氨基酸。总之,这项研究展示了CIF突出的抗炎和SS改善活性，这将为这种功能性药用植物提供进一步的见解，使其成为一种天然的抗SS药物。
