侯氏黑散化学成分对脂多糖诱导BV2细胞活化中炎性反应因子分泌平衡的影响

摘要

目的 探究侯氏黑散方中绿原酸(chlorogenic acid,CGA)、木犀草苷(cynaroside,CYN)、木犀草素(luteolin,LUT)、人参皂苷Rg1(ginsenoside Rg1,GS-Rg1)对脂多糖(lipopolysaccharide,LPS)诱导小胶质细胞炎性反应因子表达的影响.方法 实验分对照组、模型组、CGA、CYN、LUT、GS-Rg1处理组.利用LPS诱导BV2细胞过度活化,建立脑缺血后小胶质细胞炎性反应损伤体外模型;CCK-8法检测各组细胞活性;Griess法检测各组细胞上清中一氧化氮(nitric oxide,NO)的量;酶联免疫法(enzyme linked immunosorbent assay,ELISA)检测各组细胞上清中肿瘤坏死因子(tumor necrosis factor-α,TNF-α)、白介素-6(interleukin-6,IL-6)、白介素-10(interleukin-10,IL-10)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)的量;蛋白印迹法(Western blotting,WB)检测各组细胞p-p65、p-IκBα蛋白表达.结果 与模型组相比,不影响细胞生存活性的情况下,CGA、CYN、LUT、GS-Rg1可显著降低细胞上清中促炎因子NO、TNF-α、IL-6浓度;GS-Rg1可显著升高抑炎因子IL-10、TGF-β1浓度;CGA、CYN、LUT、GS-Rg1可显著降低细胞中p-p65、p-IκBα蛋白浓度.结论 CGA、CYN、LUT、GS-Rg1分别从促炎、抑炎两方面调节脑缺血后炎性反应相关因子分泌的平衡,其机制与核转录因子-κB(nuclear factor-κB,NF-κB)通路活化有关.%Objective To investigate the influence of chlorogenic acid,cynaroside,luteolin,ginsenoside Rg1 on expression of inflammatory cytokines.Methods The experiment was divided into six groups: control,model,chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 groups.The mimic ischemia injured microglia model was induced by LPS.The cyto-activity was detected via cell count kit.The NO content was determined by Griess Reagent.Contents of TNF-α、IL-6、IL-10 and TGF-β1 were detected by ELISA.The expression levels of p-p65 and p-IκBα were detected by Western blotting.Results Compared with those results of model group,chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 groups could significantly inhibit the release of NO,and decrease the content of TNF-α and IL-6,ginsenoside Rg1 markedly elevated the TGF-β1 level without influencing the cell survival.Chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 groups could decrease the expression of p-p65,p-IκBα.Conclusion The findings demonstrated that chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 played regulating roles in balancing ischemia injured microglia homeostasis via promoting anti-inflammatory cytokines as well as inhibiting the inflammatory cytokines.The therapeutic roles in the inflammatory reaction of cerebral ischemia which perhaps worked through the activation of NF-κB signaling pathway.