基础医学与临床 ›› 2017, Vol. 37 ›› Issue (11): 1535-1540.

• 研究论文 • 上一篇    下一篇

匹诺塞林缓解大鼠肝细胞系BRL-3A低氧/复氧损伤

郭振,牟童,李婷婷,朱迪,蒲俊良,吴忠均   

  1. 重庆医科大学附属第一医院
  • 收稿日期:2016-11-15 修回日期:2017-01-16 出版日期:2017-11-05 发布日期:2017-11-01
  • 通讯作者: 吴忠均 E-mail:wzjtcy@126.com
  • 基金资助:
    国家自然科学基金;重庆市科委科技人才培养计划

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

  • Received:2016-11-15 Revised:2017-01-16 Online:2017-11-05 Published:2017-11-01
  • Supported by:
    ;the grant of the Natural Science Foundation of Chongqing

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-qPCR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。

关键词: 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

Abstract: Objective To investigate the 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:the control group, the PIN group, hypoxia/reoxygenation injury group and the 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 blotting. 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 enhanced significantly, and the expression level of TLR4 and NF-κB p65 were 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 p65 were decreased while IκB-α increased. Conclusions PIN has protective effects on hypoxia/reoxygenation injury, which might be mediated in part by TLR4/NF-κB signaling pathway.

Key words: Key words:pinocembrin, hypoxia/reoxygenation, hepatocyte injury, toll-like recepcor4, nuclear factor-κb p65

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