硫酸多糖聚甘古酯抑制HIV-1反式转录调节蛋白诱导的THP-1细胞炎症细胞因子释放及机制探讨

Effect of sulfated polymannuroguluronate on Tat induced proinflammatory cytokines release in THP-1 cells and its mechanism of action

目的观察硫酸多糖聚甘古酯（SPMG）对HIV-1反式转录调节蛋白（Tat）刺激THP-1细胞释放具有神经毒性的炎症细胞因子如TNFα，IL-1β和IL-6的影响，并探讨其作用机制。方法用ELISA法检测SPMG对Tat刺激4h细胞上清液TNFα、刺激6h细胞上清液IL-1β和IL-6的影响；用Western blotting技术检测SPMG对PKCζ，PKCθ与PKCδ磷酸化影响。结果SPMG（50～100μg·mL^-1）显著抑制Tat诱导的TNFα，IL-1β与IL-6释放；Tat显著促进PKCζ，PKCθ和PKCδ的磷酸化，SPMG对PKCδ与PKCθ的磷酸化没有影响，但显著抑制PKCδ的磷酸化。结论SPMG可能通过抑制Tat对PKCδ活化，抑制炎症细胞因子TNFα，IL-6与IL-1β释放。

Aim To investigate the effects of sulfated polymannuroguluronate （SPMG）, a novel candidate anti-AIDS drug in Phase II clinical trial, on Tat-induced release of proinflammatory cytokines （i. e. , TNFα, IL-1β and IL-6） and its related mechanism. Methods The effects of SPMG on Tat induced TNFα （4 h） , IL-1β and IL-6 （6 h） secretion in THP-1 cells were measured by ELISA. Western blotting analysis was used to study the effects of SPMG on Tat induced PKCζ, PKCθ and PKCδ phosphorylation. Results SPMG （50 to 100 μg·mL^-1） markedly suppressed TNFα, IL-1β and IL-6 secretion in Tat activated THP-1 cells. In THP-1 cells the phosphorylation levels of PKCζ, PKCθ and PKCδ significantly increased following Tat stimulation, and only PKCδ phosphorylation levels was inhibited by SPMG （50 to 100 μg·mL^-1）. Conclusion SPMG suppresses the secretion of proinflammatory cytokines in THP-1 cells may be by inhibiting PKCδ activation.