Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Int J Mol Sci. 2024 Jul 5;25(13):7369. doi: 10.3390/ijms25137369.
Hyaluronidase possesses the capacity to degrade high-molecular-weight hyaluronic acid into smaller fragments, subsequently initiating a cascade of inflammatory responses and activating dendritic cells. In cases of bacterial infections, substantial quantities of HAase are generated, potentially leading to severe conditions such as cellulitis. Inhibiting hyaluronidase activity may offer anti-inflammatory benefits. Bunge, a traditional Chinese medicine, has anti-inflammatory properties. However, its effects on skin inflammation are not well understood. This study screened and evaluated the active components of that inhibit skin inflammation, using ligand fishing, enzyme activity assays, drug combination analysis, and molecular docking. By combining magnetic nanomaterials with hyaluronidase functional groups, we immobilized hyaluronidase on magnetic nanomaterials for the first time in the literature. We then utilized an immobilized enzyme to specifically adsorb the ligand; two ligands were identified as salvianolic acid B and rosmarinic acid by HPLC analysis after desorption of the dangling ligands, to complete the rapid screening of potential anti-inflammatory active ingredients in roots. The median-effect equation and combination index results indicated that their synergistic inhibition of hyaluronidase at a fixed 3:2 ratio was enhanced with increasing concentrations. Kinetic studies revealed that they acted as mixed-type inhibitors of hyaluronidase. Salvianolic acid B had K and K values of 0.22 and 0.96 μM, respectively, while rosmarinic acid had values of 0.54 and 4.60 μM. Molecular docking revealed that salvianolic acid B had a higher affinity for hyaluronidase than rosmarinic acid. In addition, we observed that a 3:2 combination of SAB and RA significantly decreased the secretion of TNF-α, IL-1, and IL-6 inflammatory cytokines in UVB-irradiated HaCaT cells. These findings identify salvianolic acid B and rosmarinic acid as key components with the potential to inhibit skin inflammation, as found in . This research is significant for developing skin inflammation treatments. It demonstrates the effectiveness and broad applicability of the magnetic nanoparticle-based ligand fishing approach for screening enzyme inhibitors derived from herbal extracts.
透明质酸酶具有将高分子量透明质酸降解为较小片段的能力，随后引发一连串炎症反应并激活树突状细胞。在细菌感染的情况下，会产生大量的透明质酸酶，可能导致蜂窝织炎等严重情况。抑制透明质酸酶的活性可能具有抗炎作用。 丹参是一种中药，具有抗炎作用。然而，其对皮肤炎症的影响尚不清楚。本研究采用配体捕捞、酶活性测定、药物组合分析和分子对接等方法，筛选和评价了 抑制皮肤炎症的活性成分。通过将磁性纳米材料与透明质酸酶的功能基团结合，我们首次在文献中实现了将透明质酸酶固定在磁性纳米材料上。然后，我们利用固定化酶特异性吸附配体；通过解吸悬挂配体后进行 HPLC 分析，鉴定出两种配体为丹酚酸 B 和迷迭香酸，从而完成 根中潜在抗炎活性成分的快速筛选。中效方程和组合指数结果表明，它们以固定的 3:2 比例协同抑制透明质酸酶，随着浓度的增加而增强。动力学研究表明，它们对透明质酸酶表现为混合型抑制剂。丹酚酸 B 的 K 和 K 值分别为 0.22 和 0.96 μM，迷迭香酸的 K 和 K 值分别为 0.54 和 4.60 μM。分子对接表明，丹酚酸 B 与透明质酸酶的亲和力高于迷迭香酸。此外，我们观察到 SAB 和 RA 的 3:2 组合可显著降低 UVB 照射的 HaCaT 细胞中 TNF-α、IL-1 和 IL-6 炎症细胞因子的分泌。这些发现表明，丹参中的丹酚酸 B 和迷迭香酸是具有抑制皮肤炎症潜力的关键成分。 本研究为开发皮肤炎症治疗方法提供了依据。它证明了基于磁性纳米颗粒的配体捕捞方法用于筛选草药提取物衍生的酶抑制剂的有效性和广泛适用性。
