摘要
目的 探讨丹酚酸B对1-甲基-4-苯基-吡啶离子(MPP+)诱导的小鼠BV2小胶质细胞神经炎症反应的影响及作用机制.方法 将BV2细胞分为空白对照组、MPP+组,以及低、中、高浓度丹酚酸B+MPP+处理组.低、中、高浓度组的细胞分别用10、50、100μmol/L丹酚酸B处理2h.此后用1 mmol/L MPP+处理细胞24 h.MPP+组仅用1 mmol/L MPP+处理24 h.采用流式细胞术和CCK-8法分别检测细胞凋亡和存活情况,采用Western blot法、免疫组化法、免疫荧光法检测各组细胞中Toll样受体4(TLR4)、髓样分化因子88(MyD88)和核因子κB(NF-κB)p65蛋白表达水平,酶联免疫吸附法测定肿瘤坏死因子α(TNF-α)、白细胞介素lβ(IL-1β)和白细胞介素6(IL-6)水平.结果 与对照组比较,MPP+组细胞TLR4、MyD88、p-NF-κB p65蛋白及炎症因子TNF-α、IL-1β和IL-6表达水平增加(P＜0.01);与MPP+组相比,低、中、高浓度丹酚酸B+MPP+处理组细胞,TLR4、MyD88、p-NF-κB p65蛋白及炎症因子TNF-α、IL-1β和IL-6表达水平与丹酚酸B浓度成正比降低(P＜0.01).结论 在MPP+诱导的小鼠炎症细胞模型中,丹酚酸B可通过抑制TLR4/NF-κB炎症信号通路激活而发挥抗炎作用.
Abstract
Objective To investigate influence and mechanism of salvianolic acid B(Sal B)on neuroinflammatory response of BV-2 microglia in mice induced by MPP+.Methods BV2 cells were divided into blank control group,MPP+group,and low,medium,and high concentration of salvianolic acid B+MPP+treatment group.Cells in the low,medium,and high concentration groups were treated with 10,50,and 100 μM salvianolic acid B for 2 hours,respectively.After that,cells were disposed with lM MPP+for 24 hours.The MPP+group was merely treated with IM MPP+for 24 hours.Flow cytometry and CCK-8 assay were used to detect cellular apoptosis and survival status.Western blot,immunohistochemistry,and immunofluorescence were utilized to detect expression levels of proteins such as toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor κB(NF-κB)p65 in each group of cells.Enzyme-linked immunosorbent assay was applied to measure expression levels of tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β),and interleukin 6(IL-6).Results Compared with the control group,cellular expression levels of proteins including TLR4,MyD88 and p-NF-KB p65,as well as inflammatory factors containing TNF-α,IL-1 β and IL-6 in the MPP+group increased(P＜0.01).In comparision with the MPP+group,cellular expression levels of proteins including TLR4,MyD88 and p-NF-κB p65,as well as inflammatory factors containing TNF-α,IL-1 β and IL-6 in the group treated with low,medium,and high concentrations of salvianolic acid B+MPP+were downregulated in direct proportion to the concentration of salvianolic acid B(P＜0.01).Conclusion In the MPP+-induced mouse inflammatory cell model,salvianolic acid B exerted anti-inflammatory effect by inhibiting activation of the TLR4/NF-κB inflammatory signaling pathway.
基金项目
甘肃省卫生行业科研计划项目(GSWSKY2018-43)
甘肃省科技计划项目(21JR7RA008)
NETL
NSTL
万方数据