曲古霉素A对CD14+单核细胞活化及TLR4信号通路的影响

目的: 研究曲古霉素A（trichostatin A,TSA）对CD14⁺单核细胞活化状态及Toll样受体4（TLR4）炎性信号通路的影响及其具体机制。方法: 分离健康人外周血CD14⁺单核细胞，用20 nmol/L浓度TSA孵育24 h后收集细胞，RT-qPCR方法检测TLR4及下游炎症因子基因mRNA表达水平，以流式细胞分析检测细胞表面TLR4蛋白表达量，以ChIP-qPCR方法检测TLR4基因启动子区组蛋白H3乙酰化水平变化，以Transwell法检测单核细胞趋化能力，以细胞黏附力实验检测单核细胞的黏附能力。结果: TSA刺激能够增加CD14+单核细胞的趋化和黏附能力，上调CD14⁺单核细胞中TLR4及下游炎症因子TNF-α、MCP-1和IL-6的表达，提高TLR4基因启动子区组蛋白H3乙酰化水平。结论: TSA能够诱导CD14⁺单核细胞活化，提高TLR4启动子区组蛋白乙酰化修饰水平，促进TLR4信号通路基因过度表达。

Effects of trichostatin A on CD14+ monocytes activation and TLR4 signal pathway

AIM: To study the effects of trichostatin A (TSA) on CD14⁺ monocytes activation and TLR4 signal pathway. METHODS: CD14⁺ monocytes were isolated from healthy donors and cultured in 1640 medium with 20 nmol/L TSA. After 24 hours, cells were harvested to detect the expression of TLR4 and downstream cytokines by RT-qPCR and flow cytometry methods. ChIP-qPCR was used to detect histone H3 acetylation level TLR4 promoter. Transwell and adhesion experiments were performed to assess the motility and adhesion ability of CD14⁺ monocytes. RESULTS: TSA treatment can increase the motility and adhesion ability of CD14⁺ monocytes. Expression of TLR4 and down-stream cytokines, including TNF-α, MCP-1 and IL-6, were up-regulated in CD14⁺ monocytes after TSA treatment. Higher acetylation of histone H3 within TLR4 promoter region was also detected in CD14⁺ monocytes after TSA treatment. CONCLUSION: TSA activates CD14⁺ monocytes, promotes the expression of genes in TLR4 signal pathway and increases the histone H3 acetylation of TLR4 promoter.