Zou Caomin, Chen Qianru, Li Jiasheng, Lin Xiguang, Xue Xingyang, Cai Xinhang, Chen Yicheng, Sun Yue, Wang Shumei, Zhang Ying, Meng Jiang
School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica, State Administration of Traditional Chinese Medicine (TCM), Engineering Technology Research Center for Chinese Materia Medica Quality of Universities in Guangdong Province, Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization, Guangzhou, China.
Affiliated Cancer Hospital and Institute of Guangzhou Medical University, Guangzhou, China.
Front Pharmacol. 2024 Feb 7;15:1358640. doi: 10.3389/fphar.2024.1358640. eCollection 2024.
Moutan Cortex (MC) has been used in treating inflammation-associated diseases and conditions in China and other Southeast Asian countries. However, the active components of its anti-inflammatory effect are still unclear. The study aimed to screen and identify potential cyclooxygenase-2 (COX-2) inhibitors in MC extract. The effect of MC on COX-2 was determined by COX-2 inhibitory assays, followed by bio-affinity ultrafiltration in combination with ultra-performance liquid chromatography-mass spectrometry (BAUF-UPLC-MS). To verify the reliability of the constructed approach, celecoxib was applied as the positive control, in contrast to adenosine which served as the negative control in this study. The bioactivity of the MC components was validated by COX-2 inhibitor assay and RAW264.7 cells. Their anti-inflammatory activity was also evaluated using LPS-induced zebrafish inflammation models. Finally, molecular docking was hired to further explore the internal interactions between the components and COX-2 residues. The MC extract showed an evident COX-2-inhibitory effect in a concentration-dependent manner. A total of 11 potential COX-2 inhibitors were eventually identified in MC extract. The COX-2 inhibitory activity of five components, namely, gallic acid (GA), methyl gallate (MG), galloylpaeoniflorin (GP), 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG), and 1,2,3,4,6-Penta-O-galloyl-β-D-glucopyranose (PGG), were validated through both assays and experiments using zebrafish models. Besides, the molecular docking analysis revealed that the potential inhibitors in MC could effectively inhibit COX-2 by interacting with specific residues, similar to the mechanism of action exhibited by celecoxib. In conclusion, BAUF-UPLC-MS combining the molecular docking is an efficient approach to discover enzyme inhibitors from traditional herbs and understand the mechanism of action.
在中国和其他东南亚国家，牡丹皮已被用于治疗炎症相关疾病和病症。然而，其抗炎作用的活性成分仍不清楚。本研究旨在筛选和鉴定牡丹皮提取物中潜在的环氧化酶-2（COX-2）抑制剂。通过COX-2抑制试验测定牡丹皮对COX-2的作用，随后结合超高效液相色谱-质谱联用技术（BAUF-UPLC-MS）进行生物亲和超滤。为验证所构建方法的可靠性，本研究将塞来昔布作为阳性对照，腺苷作为阴性对照。通过COX-2抑制剂试验和RAW264.7细胞验证了牡丹皮成分的生物活性。还使用脂多糖诱导的斑马鱼炎症模型评估了它们的抗炎活性。最后，采用分子对接进一步探索成分与COX-2残基之间的内在相互作用。牡丹皮提取物呈浓度依赖性地显示出明显的COX-2抑制作用。最终在牡丹皮提取物中鉴定出11种潜在的COX-2抑制剂。通过试验和斑马鱼模型实验验证了五种成分，即没食子酸（GA）、没食子酸甲酯（MG）、没食子酰芍药苷（GP）、1,2,3,6-四-O-没食子酰-β-D-葡萄糖（TGG）和1,2,3,4,6-五-O-没食子酰-β-D-吡喃葡萄糖（PGG）的COX-2抑制活性。此外，分子对接分析表明，牡丹皮中的潜在抑制剂可通过与特定残基相互作用有效抑制COX-2，类似于塞来昔布的作用机制。总之，BAUF-UPLC-MS结合分子对接是从传统草药中发现酶抑制剂并了解其作用机制的有效方法。
Zotero 插件
