摘要
目的 研究新型香豆素衍生物P492B对小胶质细胞介导的神经炎症的作用.方法 以香豆素为母核,合成新型香豆素衍生物P95B、P613B 和P492B.通过MTT法评估P95B、P613B 和P492B对SH-SY5Y神经元细胞系细胞活力的影响,筛选最优化合物P492B.构建脂多糖(LPS)诱导的BV-2 小胶质细胞神经炎症模型,给予P492B处理后,形态学观察P492B对小胶质细胞活化的影响.Western blot法检测P492B对小胶质细胞释放一氧化氮合成酶(iNOS)蛋白水平的影响.ELISA 法检测P492B 对小胶质细胞释放的炎症因子TNF-α和IL-6 水平的影响,DCFH-DA荧光探针检测P492B对小胶质细胞ROS水平的影响.结果 香豆素、P95B和P613B均可剂量依赖性降低神经细胞活力,而P492B对神经细胞活力无显著影响.P492B可显著改善小胶质细胞的活化,并剂量依赖性的显著降低活化的小胶质细胞iNOS蛋白水平、剂量依赖性的显著减少炎症因子TNF-α和IL-6 的表达水平、剂量依赖性的显著降低活化的小胶质细胞活性氧水平.结论 新型香豆素衍生物P492B可显著抑制小胶质细胞介导的神经炎症,为神经炎症相关疾病的治疗提供新的潜在治疗药物.
Abstract
Objective To investigate the effects of a novel coumarin derivative,P492B,on the microglia-mediated neuroinflammation.Methods Using coumarin as the core structure,new coumarin derivatives P95B,P613B,and P492B were synthesized.The effect of these derivatives on the viability of SH-SY5Y neuronal cells was evaluated using MTT assay,and the optimal compound,P492B,was selected.Next,the neuroinflammation model induced by LPS in BV-2 microglia was established,and the effect of P492B on microglial activation was analyzed through morphological observations.Moreover,the effect of P492B on the expression of iNOS protein in BV-2 microglia was assessed using Western blot analysis.The levels of inflammatory factors tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)released by BV-2 microglia were measured using ELISA.The effect of P492B on the level of reactive oxygen species(ROS)in BV-2 microglia was determined using the DCFH-DA fluorescent probe.Results The cell viability of SH-SY5Y cells was significantly reduced in a dose-dependent manner by,P95B,and P613B,while P492B had no significant effect on the cell viability of SH-SY5Y cells.The microglia activation was significantly inhibited by P492B,and the protein level of iNOS of activated microglia was significantly reduced by P492B in dose-dependent manner.The levels of inflammatory factors TNF-α and IL-6 were significantly decreased by P492B in dose-dependent manner and the ROS levels in activated microglia was significantly reduced by P492B.Conclusion The novel coumarin derivative,P492B,inhibits microglia-mediated neuroinflammation,which provids new insights and potential therapeutic options for neuroinflammatory-related diseases.
基金项目
广西壮族自治区药品监督管理局药品安全科研项目(桂药监科直属[2023]008号)
维普
万方数据