PDF(Pubmed)
Abstract：
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. Salvia miltiorrhiza 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 S. miltiorrhiza 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 S. miltiorrhiza 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 Ki and Kis 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 S. miltiorrhiza. 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.
摘要：
透明质酸酶具有将高分子量透明质酸降解成较小片段的能力，随后启动炎症反应的级联反应和激活树突状细胞。在细菌感染的情况下，产生大量的HAase，可能导致严重的疾病，如蜂窝织炎。抑制透明质酸酶活性可提供抗炎益处。丹参,中药，具有抗炎特性。然而,它对皮肤炎症的影响尚不清楚。本研究筛选并评价了丹参抑制皮肤炎症的活性成分,使用配体捕鱼，酶活性测定，药物组合分析，和分子对接。通过将磁性纳米材料与透明质酸酶官能团结合，我们在文献中首次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；在悬空配体解吸后，通过HPLC分析鉴定了两个配体为丹酚酸B和迷迭香酸，完成丹参根中潜在抗炎活性成分的快速筛选。中值效应方程和组合指数结果表明，在固定的3：2比率下，它们对透明质酸酶的协同抑制作用随着浓度的增加而增强。动力学研究表明，它们充当透明质酸酶的混合型抑制剂。丹酚酸B的Ki和Kis值为0.22和0.96μM，分别,而迷迭香酸的值为0.54和4.60μM。分子对接显示丹酚酸B对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组合显着降低TNF-α的分泌，UVB照射的HaCaT细胞中的IL-1和IL-6炎性细胞因子。这些发现确定丹酚酸B和迷迭香酸是具有抑制皮肤炎症潜力的关键成分，如在丹参中发现的。这项研究对于开发皮肤炎症治疗具有重要意义。它证明了基于磁性纳米颗粒的配体打捞方法用于筛选源自草药提取物的酶抑制剂的有效性和广泛适用性。
