在湿性年龄相关性黄斑变性中Toll样受体系统对补体系统的作用

Effects of Toll-like receptors on complement system in patients with wet-age-related macular degeneration

摘要:

目的：探讨在湿性年龄相关性黄斑变性（age-related macular degeneration，AMD）病人中Toll样受体（toll like receptor，TLR）2、3对补体C3a、C5a的作用。方法：收集15例湿性AMD病人及15例正常人外周血，采用梯度离心法分离出外周血单个核细胞（peripheral blood mononuclear cells，PBMCs），将细胞分为空白组、C3a组、C5a组、TLR2作用C3a组、TLR2作用C5a组、TLR3作用C3a组、TLR3作用C5a组共7个组并分别给予对应刺激。收集上清液，采用ELISA法检测AMD相关细胞因子白细胞介素（interleukin，IL）-1β、IL-6、IL-8、IL-17、单核细胞趋化蛋白-1（monocyte chemoattractant protein-1，MCP-1）、血管内皮生长因子（vascular endothelial growth factor，VEGF）的含量。结果：正常人及湿性AMD病人中，TLR2作用C3a组、TLR2作用C5a组、TLR3作用C3a组、TLR3作用C5a组上清液中IL-1β、IL-6、IL-8、MCP-1的量均高于C3a组或C5a组；湿性AMD病人中，空白组、C3a组、C5a组、TLR2作用C3a组、TLR2作用C5a组、TLR3作用C3a组、TLR3作用C5a组上清液中IL-1β、IL-6、IL-8、MCP-1的量较正常人对应组明显升高。结论：Toll样受体系统对补体系统的作用有可能促进了湿性AMD的发生发展。

Abstract:

Objective：To explore the effects of Toll-like receptors（TLR2，TLR3） on C3a and C5a in patients with wet-age-related macular degeneration（wet-AMD）. Methods：Peripheral blood samples were collected from 15 patients with wet-AMD and 15 subjects without wet-AMD were taken as controls. Peripheral blood mononuclear cells were then isolated using gradient centrifugation. The iso－lated cells were divided into 7 groups：blank group，C3a group，C5a group，TLR2 on C3a group，TLR2 on C5a group，TLR3 on C3a group and TLR3 on C5a group. Stimulations were added to each group. The supernatant was collected and tested for concentration of AMD-related cytokines，such as interleukin-1Beta，interleukin-6，interleukin-8，interleukin-17，monocyte chemoattractant protein-1，vascular endothelial growth factor by using ELISA. Results：The concentrations of interleukin-1Beta，interleukin-6，interleukin-8 and monocyte chemoattractant protein-1 were higher in TLR2 on C3a group，TLR2 on C5a group，TLR3 on C3a group and TLR3 on C5a group than in C3a group or C5a group. Concentrations of interleukin-1beta，interleukin-6，interleukin-8 and monocyte chemoattractant protein-1 were obviously increased in blank group，C3a group，C5a group，TLR2 on C3a group，TLR2 on C5a group，TLR3 on C3a group and TLR3 on C5a group of patients with wet-AMD than in corresponding group of subjects without wet-AMD. Conclusion：The effects of Toll-like receptors on the complement system may promote the development of wet-AMD.