枸杞多糖对同型半胱氨酸致THP-1单核细胞源性巨噬细胞脂质沉积及内质网应激的影响

摘要

目的 探讨枸杞多糖( LBP)对同型半胱氨酸( Hcy)致THP-1单核细胞源性巨噬细胞脂质沉积及内质网应激( ERS)的影响. 方法 复制THP-1单核细胞源性巨噬细胞模型,共分为以下6组:NC组不做其他处理,Hcy组选用100μmol/L的Hcy干预,20 LBP组选用终浓度100μmol/L的Hcy+20μg/mL的LBP干预,50 LBP组选用终浓度100μmol/L的Hcy+50μg/mL的LBP干预,100 LBP组选用终浓度100μmol/L的Hcy+100μg/mL的LBP干预,200 LBP组选用终浓度100μmol/L的Hcy+200μg/mL的LBP干预. 显微镜下观察巨噬细胞模型是否复制成功;RT-qPCR分析脂肪酸结合蛋白4 ( FABP4 ) mRNA表达改变;免疫荧光检测巨噬细胞内FABP4蛋白含量的变化;ELISA检测细胞内氧化低密度脂蛋白(ox-LDL)及内质网应激标志蛋白(CHOP、GRP78、XBP-1)含量的变化. 结果 成功复制THP-1单核细胞源性巨噬细胞模型. 与NC组比较,Hcy组巨噬细胞内ox-LDL及FABP4的含量及内质网应激标志蛋白表达显著增加(P<0.05,P<0.01). 而不同LBP作用浓度组间比较,ox-LDL含量和FABP4 mRNA水平,50LBP组、100LBP组、200LBP组与Hcy组比较,差异有统计学意义( P<0.05,P<0.01),100LBP组、200LBP组与50LBP组比较差异有统计学意义(P<0.05). 200 LBP组与Hcy组、50 LBP组比较,GRP78含量差异统计学意义( P<0.05,P<0.01). 100 LBP组、200 LBP组与Hcy组比较,CHOP含量差异有统计学意义(P<0.01),200 LBP组与50 LBP组比较,CHOP含量差异有统计学意义(P<0.05). 50 LBP组、100 LBP组、200 LBP组与Hcy组比较,Xbp-1含量差异有统计学意义( P<0.01 ) ,100 LBP组、200 LBP组与50 LBP组比较,Xbp-1含量差异有统计学意义(P<0.01). 结论 随着LBP浓度的增加,可逐渐减少Hcy导致的THP-1单核细胞源性巨噬细胞内的脂质沉积及内质网应激蛋白含量,从而发挥抗动脉粥样硬化的作用.%Objective To investigate the effect of lycium barbarum polysaccharide ( LBP ) on homocysteine ( Hcy) -induced lipid accumulation and endoplasmic reticulum stress in THP -1 monocyte -derived macrophages. Methods The model of THP-1 monocyte-derived macrophages was replicated.Cultured cells were divided into 6 groups:the control group (NC), the 100 μmol/L Hcy group (Hcy), the 100 μmol/L Hcy+20, 50,100, 200 μg/mL LBP groups.The macrophage model was verified under microscope.Fatty acid-binding protein 4 ( FABP4) mRNA levels were detected by real-time PCR.FABP4 protein levels in macrophages were analyzed by immunofluorescence.Intracellu-lar oxygenized low density lipoprotein (ox-LDL) and endoplasmic reticulum stress markers (CHOP, GRP78, XBP-1) levels were measured by ELISA kits.Results The macrophage model was successfully established.Treatments of 100μmol/L Hcy and LBP at various concentrations showed that levels of ox-LDL, FABP4 and endoplasmic reticulum stress markers were higher in the Hcy group than in the NC group.While different levels of LBP diminished the alterations in an apparent dose-dependent manner.Conclusion At increasing concentrations, LBP attenuates homocysteine-induced lipid accumulation and down-regulates the levels of endoplasmic reticulum stress markers, thus playing a role of anti-atherosclerosis.