匹诺塞林缓解大鼠肝细胞系BRL-3A低氧/复氧损伤  

Pinocembrin alleviates hypoxia reoxygenation injury in rat liver cell line BRL-3A

作　　者：郭振[1] 牟童[1] 李婷婷[1] 朱迪[1] 蒲俊良 吴忠均[1] 

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆400016

出　　处：《基础医学与临床》2017年第11期1535-1540,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81672959);重庆市科委科技人才培养计划(cstc2015shmszx120019)

摘　　要：目的探讨匹诺塞林(PIN)对低氧/复氧(H/R)诱导的大鼠肝细胞损伤的保护作用及其可能机制。方法将大鼠肝细胞(BRL-3A细胞系)分为正常对照组、PIN实验组、低氧复氧损伤模型组和PIN预处理组。CCK-8检测细胞存活率;Annexin V-FITC/PI双染法流式细胞计量术检测细胞凋亡;检测细胞培养液中谷丙转氨酶(ALT)活性;ELISA检测TNF-α和IL-1β含量;Western blot检测细胞中TLR4、IκB-α和NF-κB P65蛋白水平;RT-q PCR检测细胞中TLR4、IκB-α和NF-κB P65mRNA表达。结果在H/R条件下细胞存活率明显降低(P<0.01),细胞凋亡率增高(P<0.001),ALT活性升高(P<0.01),IL-1β和TNF-α含量增多(P<0.01),TLR4和NF-κB P65蛋白与mRNA表达水平显著提高(P<0.01)而IκB-α降低(P<0.05);经PIN预处理后,细胞存活率显著提高(P<0.01),细胞凋亡率显著减小(P<0.001),ALT活性降低(P<0.01),IL-1β和TNF-α含量降低(P<0.01),TLR4和NF-κB P65蛋白与mRNA表达水平明显降低(P<0.01)而IκB-α升高(P<0.05)。结论 PIN对H/R诱导的BRL-3A肝细胞的损伤具有保护作用,且该作用可能是通过TLR4/NF-κB信号通路实现的。Objective To investigate protective effect of pinocembrin（PIN） on hepatocytes induced by hypoxia/reoxygenation（H/R） as well as its relationship to the TLR4/NF-κB signaling pathway.Methods The cells were randomly divided into 4 groups： control group,PIN group,hypoxia/reoxygenation injury group and PIN pretreatment group.The cell viability was measured with CCK-8.The apoptosis rate was determined by Annexin V-FITC/PI staining.The activity of ALT was detected.ELISA was used to evaluate the contents of TNF-α and IL-β.The mRNA and protein expression level of TLR4,IκB-α and NF-κB P65 in cells was observed by quantitative real-time PCR or Western blot.Results The H/R stimulation decreased cell survival rate and enhanced the apoptosis.The activity of ALT was increased.The contents of TNF-α and IL-1β were significantly enhanced,and the expression level of TLR4 and NF-κB P65 was markedly increased while IκB-α decreased.After pretreatment with PIN,the cell survival rate increased while the apoptosis rate decreased.The activity of ALT was decreased.TNF-α and IL-1β were reduced significantly and the expression level of TLR4 and NF-κB P 65 were decreasedwhile IκB-α increased.Conclusions PIN has protective effects on hypoxia/reoxygenation injury,which might be mediated in part by TLR4/NF-κB signaling pathway.

关 键 词：匹诺塞林 低氧/复氧 肝细胞损伤 Toll样受体4 核转录因子-ΚB P65 

分 类 号：R965[医药卫生—药理学]