黄芪汤调节TLR4/NF-κB信号通路改善高糖诱导足细胞损伤的研究

Huangqi Decoction Regulates TLR4/NF-κB Pathway to Improve High Glucose-Induced Podocyte Injury

    摘要
  • 摘要:
      目的  探讨黄芪汤对高糖诱导足细胞损伤的作用及机制。
      方法  体外培养人足细胞, 采用30 mmol·L-1葡萄糖干预24 h诱导足细胞损伤, 分别设置对照组、模型组、黄芪汤低浓度组(10 μg·mL-1)、黄芪汤中浓度组(30 μg·mL-1)和黄芪汤高浓度组(100 μg·mL-1)。采用CCK-8法检测细胞增殖能力, 划痕实验检测细胞迁移能力，qPCR法检测TNF-α、IL-6等炎症因子mRNA表达, ELISA法检测足细胞上清TNF-α、IL-6的含量，Western blot法检测足细胞TLR4、NF-κB、p-NF-κB、TNF-α及IL-6蛋白表达。
      结果  与对照组比较, 模型组细胞划痕愈合率明显降低(P < 0.01)，足细胞中TNF-α、IL-6、CCL24 mRNA表达水平，上清液中TNF-α、IL-6的含量和TLR4、p-NF-κB/NF-κB、TNF-α及IL-6蛋白表达均显著增加(P < 0.05，P < 0.01)。与模型组相比, 黄芪汤组细胞划痕率明显升高(P < 0.01)，足细胞中TNF-α、IL-6的含量和mRNA表达，以及TLR4、p-NF-κB/NF-κB、TNF-α、IL-6蛋白表达均显著减少(P < 0.05, P < 0.01)。
      结论  黄芪汤能有效减轻高糖对人足细胞的炎症损伤, 增强细胞增殖、迁移能力, 抑制细胞凋亡, 其机制可能与抑制TLR4/NF-κB信号通路、下调炎症因子的表达有关。

     

    Abstract:
      OBJECTIVE  To investigate the effect and mechanism of Huangqi Decoction on high glucose-induced podocyte injury.
      METHODS  Human podocytes were cultured in vitro and treated with 30 mmol·L-1 glucose for 24 h to induce injury. The groups were divided into control group, model group, low-, medium- and high-concentration of Huangqi Decoction groups (10, 30, 100 μg·mL-1). The cell proliferation was detected by CCK-8 method, the mRNA expression of TNF-α and IL-6 was detected by qPCR, the contents of TNF-α and IL-6 in podocyte supernatant were detected by ELISA, and the protein expression of TLR4, p-NF-κB, NF-κB, TNF-α and IL-6 in podocytes was detected by Western blot.
      RESULTS  Compared with the control group, the mRNA expression of TNF-α, IL-6, CCL24, the contents of TNF-α and IL-6 and the protein expression of TLR4, p-NF-κB/NF-κB, TNF-α and IL-6 in podocytes significantly increased (P < 0.05, P < 0.01). Compared with the model group, the mRNA expression and contents of TNF-α, IL-6 and the protein expression of TLR4, p-NF-κB/NF-κB, TNF-α and IL-6 in podocytes of Huangqi Decoction groups significantly decreased (P < 0.05, P < 0.01).
      CONCLUSION  Huangqi Decoction can reduce the inflammatory damage of human podocytes induced by high glucose, enhance cell proliferation and migration, and inhibit cell apoptosis. The mechanism may be related to the inhibition of TLR4/NF-κB signal pathway and down-regulation of the inflammatory factors expression.

     

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