天津医药

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雷公藤红素抑制磨损颗粒诱导细胞炎症反应的研究

李哲1,江川1,戴闽1,邹飏2,3,刘翔2,3,尚江荫子2,3   

  1. 1. 南昌大学第一附属医院骨科
    2.
    3. 南昌大学第一附属医院
  • 收稿日期:2013-01-09 修回日期:2013-05-21 出版日期:2013-09-15 发布日期:2013-09-15
  • 通讯作者: 戴闽

Inhibitory Effects of Tripterine on Wear Particle-Induced Cellular Inflammatory Reaction

LI Zhe 1,JIANG Chuan 1,DAI Min 1,ZOU Yang, 1,2,LIU Xiang 1,2,SHANG Jiang yinzi 1   

  • Received:2013-01-09 Revised:2013-05-21 Published:2013-09-15 Online:2013-09-15
  • Contact: DAI Min

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摘要：

【摘要】目的  通过体外细胞培养，探讨雷公藤红素抑制人工关节磨损颗粒诱导的无菌性炎症反应的效果。方法  使用真空球磨法制备人工关节磨损颗粒并用无血清培养基配制颗粒悬液。使用RAW264.7巨噬细胞进行传代培养，并将其按不同处理因素随机分成4组：空白对照组（A组）、磨损颗粒组（B组）、磨损颗粒+雷公藤红素组（C组）和雷公藤红素组（D组）。分组处理24h后行CCK-8毒性检测；通过ELISA检测各组肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β表达情况；RT-PCR检测TNF-α、IL-1β和核因子（NF）-κB的基因表达量；Western Blot在蛋白水平检测NF-κB的表达情况。结果  1mg/L雷公藤红素的细胞毒性作用不明显；磨损颗粒组促炎症因子TNF-α、IL-1β的表达和NF-κB的激活明显高于空白对照组（P＜0.05），加入雷公藤红素后以上各指标表达均明显降低（P＜0.05）。结论    雷公藤红素可在基因和蛋白水平抑制磨损颗粒诱导RAW264.7细胞促炎症因子的表达，并可抑制NF-κB通路
的激活。

关键词: 雷公藤属, 关节, 巨噬细胞, 炎症, 肿瘤坏死因子α, 白细胞介素1β, NF-κB, 雷公藤红素

Abstract: [Abstract]  Objective   To investigate the effect of tripterine on wear particle-induced inflammatory reaction by cell culture in vitro. Methods  Wear particles from artificial joints were prepared by vacuum ball milling, and made into particle suspension by using serum-free medium. RAW264.7 cells were cultured, subcultured and divided randomly into four groups according to different treatment factors: blank control group (group A), wear particle group (group B), wear particle + tripterine group (group C) and tripterine group (group D). After 24 hours, toxicity of tripterine was detected by CCK-8 assay; ELISA was used to detect the expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) ; RT-PCR was used to detect the expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and nuclear factor-κB (NF-κ B) at the level of gene;Western blot was used to detect the expression of nuclear factor-κB (NF-κ B) at the level of protein. Results  The results showed 1μg/ml tripterine was little cytotoxic；the expressions of pro-inflammatory cytokines and the activation of NF-κB in wear particle group were significantly higher than those in blank control group (P＜0.05)，and the indexes after the treatment with tripterine (group C)decreased statistically (P＜0.05). Conclusion  Tripterine can inhibit the wear particle-induced expression and release of pro-inflammatory cytokines at the level of gene and protein and the activation of NF-κB in RAW264.7 cells.

Key words: tripterygium, joints, macrophages, inflammation, tumor necrosis factor-alpha, interleukin-1beta, NF-kappa B, 雷公藤红素