二氢杨梅素缓解神经炎症作用及机制研究
首发时间:2022-05-07
摘要:目的:二氢杨梅素(DHM)是从显齿葡萄中提取的黄酮类化合物。本课题旨在探讨DHM在小胶质细胞炎性活化中的作用及机制,为治疗神经炎症相关疾病提供基础。方法:Griess试剂检测NO的释放,MTT检测细胞活力。qPCR和Western-blot检测炎症因子表达。Western-blot和免疫荧光检测NF-κB信号通路的激活。划痕实验和transwell评价小胶质细胞的迁移趋化。使用DCFH-DA和MitoSOX检测ROS和mtROS的产生。最后,通过小胶质细胞条件培养液/神经元共培养实验探讨DHM对神经元的保护作用。结果:DHM剂量依赖性地抑制M1型小胶质细胞炎症介质的产生以及迁移趋化反应。此外,DHM显著抑制ROS及mtROS的产生。最后,DHM能减缓小胶质细胞对神经元的炎性损伤,从而发挥神经保护作用。结论:DHM抑制小胶质细胞过度激活,从而减缓神经元炎性损伤,起到神经保护作用。
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Anti-inflammatory effect of Dihydromyricetin on microglial activation
Abstract:Objective:Dihydromyricetin (DHM) is a flavonoid compound extracted from dentine grape. Our study explored the role and mechanism of DHM in inflammatory activation of microglia in order to provide a basis for drugs discovery for neuroinflammation-related diseases. Method:NO release was detected by Griess reagent, the cell viability was measured by MTT method, the mRNA and protein expression of inflammatory factors were detected by q-PCR and Western-blot. And the activation of NF-κB signaling pathway was detected by immunoblotting and immunofluorescence assay. Then, the migration and chemotaxis of microglia were measured by wound healing assays and transwell assays. Furthermore, The production of ROS and mtROS was detected by DCFH-DA and MitoSOX.Finally, the neurons protective effect of DHM was observed in neuron/microglial co-culture system. Results:The results showed that DHM inhibited the production of M1 microglia inflammatory mediators in dose-dependent manner, such as TNFα, IL-6, IL-1β, COX-2, iNOS and NO. And DHM also inhibited migration chemotactic response of M1 microglia. Furthermore, DHM significantly inhibit the production of ROS. Finally,DHM has been shown to the neurons protective effect by reducing microglia inflammatory response in neuron/microglial co-culture system. Conclusion: In this study, our results suggest that DHM inhibits microglia over-activation, so as to decrease neuron loss.
Keywords: Dihydromyricetin microglia neuroinflammation
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