Lee I-Ta, Lin Horng-Chyuan, Huang Tse-Hung, Tseng Chi-Nan, Cheng Hao-Tsa, Huang Wen-Chung, Cheng Ching-Yi
School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei 11031, Taiwan.
Department of Thoracic Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan 33305, Taiwan; College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan.
Antioxidants (Basel). 2022 Apr 25;11(5):835. doi: 10.3390/antiox11050835.
In Taiwan, the root extract of Sieb. et Zucc. (Vitaceae, VT) is rich in stilbenes, with resveratrol (Res) and its derivatives being the most abundant. Previously, we showed that the effect of Res derivatives against tumor necrosis factor-α (TNF-α)-stimulated inflammatory responses occurs via cPLA2/COX-2/PGE2 inhibition. This study compared and explored the underlying anti-inflammatory pharmacological mechanisms. Before stimulation with TNF-α, RMCs were treated with/without pharmacological inhibitors of specific protein kinases. The expression of inflammatory mediators was determined by Western blotting, gelatin zymography, real-time PCR, and luciferase assay. Cellular and mitochondrial ROS were measured by HDHFDA or DHE and MitoSOX™ Red staining, respectively. The RNS level was indirectly measured by Griess reagent assay. Kinase activation and association were assayed by immunoprecipitation followed by Western blotting. TNF-α binding to TNFR recruited Rac1 and p47, thus activating the NAPDH oxidase-dependent MAPK and NF-κB pathways. The TNF-α-induced NF-κB activation via c-Src-driven ROS was independent from the EGFR signaling pathway. The anti-inflammatory effects of Res derivatives occurred via the inhibition of ROS derived from mitochondria and NADPH oxidase; RNS derived from iNOS; and the activation of the ERK1/2, JNK1/2, and NF-κB pathways. Overall, this study provides an understanding of the various activities of Res derivatives and their pharmacological mechanisms. In the future, the application of the active molecules of VT to health foods and medicine in Taiwan may increase.
在台湾，蛇葡萄（葡萄科，VT）的根提取物富含芪类化合物，其中白藜芦醇（Res）及其衍生物含量最为丰富。此前，我们发现Res衍生物对肿瘤坏死因子-α（TNF-α）刺激的炎症反应的作用是通过抑制cPLA2/COX-2/PGE2实现的。本研究比较并探讨了其潜在的抗炎药理机制。在用TNF-α刺激之前，将大鼠系膜细胞（RMCs）用特定蛋白激酶的药理抑制剂处理或不处理。通过蛋白质免疫印迹法、明胶酶谱法、实时聚合酶链反应（PCR）和荧光素酶测定法测定炎症介质的表达。分别通过HDHFDA或二氢乙锭（DHE）以及MitoSOX™ Red染色测量细胞和线粒体活性氧（ROS）。通过格里斯试剂测定法间接测量活性氮（RNS）水平。通过免疫沉淀后进行蛋白质免疫印迹法测定激酶激活和关联。TNF-α与肿瘤坏死因子受体（TNFR）结合募集Rac1和p47，从而激活依赖烟酰胺腺嘌呤二核苷酸磷酸（NAPDH）氧化酶的丝裂原活化蛋白激酶（MAPK）和核因子κB（NF-κB）途径。TNF-α通过c-Src驱动的ROS诱导的NF-κB激活独立于表皮生长因子受体（EGFR）信号通路。Res衍生物的抗炎作用是通过抑制线粒体和NADPH氧化酶产生的ROS、诱导型一氧化氮合酶（iNOS）产生的RNS以及激活细胞外信号调节激酶1/2（ERK1/2）、应激活化蛋白激酶1/2（JNK1/2）和NF-κB途径实现的。总体而言，本研究有助于了解Res衍生物的多种活性及其药理机制。未来，VT活性分子在台湾保健食品和药物中的应用可能会增加。
