机械牵拉对培养人视网膜色素上皮细胞MCP-1和IL-8表达的影响  

Effects of mechanical stress on expressions of monocyte chemoattractant protein-1 and interleukin-8 of cultured human retinal pigment epithelial cells

作　　者：张晓光[1] 惠延年[1] 韩泉洪[1] 侯旭[1] 陈立军[1] 马吉献[1] 

机构地区：[1]第四军医大学西京医院全军眼科研究所,西安710032

出　　处：《中华医学杂志》2005年第32期2264-2268,共5页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30200311)

摘　　要：目的模拟和了解培养的人视网膜色素上皮细胞(hRPE)单核细胞趋化蛋白1(MCP1)和白介素8(IL8)的表达,可能在原发性视网膜脱离形成早期的作用。方法将胶原包被的磁珠悬液加入培养的hRPE细胞,孵育并洗去未结合的磁珠。用逆转录聚合酶链反应和酶联免疫反应分别测定hRPE细胞在磁场垂直方向力作用下,以及在细胞松弛素D处理后,各时间点的MCP1和IL8的mRNA和培养液上清中的蛋白水平。结果不受磁场作用的hRPE表达极低的MCP1及IL8。在牵拉力作用下,MCP1及IL8的表达有2个峰值。其mRNA的第1个峰值出现在0.5h内,分别达到3.30ng/L±0.12ng/L和1.88ng/L±0.08ng/L。蛋白水平的第1个峰值出现在1h内,分别为552.05ng/L±7.64ng/L和236.67ng/L±14.30ng/L。第2个峰值出现在4h左右。细胞松弛素D预处理后两种因子的表达和分泌量均明显降低,分别为2.36ng/L±0.27ng/L和1.64ng/L±0.08ng/L,以及353.80ng/L±16.68ng/L和101.86ng/L±15.92ng/L。结论机械牵拉力可诱导hRPE细胞表达MCP1和IL8,这种作用在细胞松弛素D预处理后出现部分抑制,提示其影响有细胞骨架的参与,这两种炎性因子在视网膜脱离发生的起始阶段起到一定作用。Objective To investigate the expressions of monocyte chemoattractant protein-1 （ MCP- 1 ） and interleukin-8 （IL-8） of cultured human retinal pigment epithelial （hRPE） cells under mechanical stress in vitro, so as to mimic and understand the role of these 2 inflammatory cytokines in the early stage of development of primary retinal detachment. Methods Ferric oxide beads coated with collagen were added to the culture plate containing waU-attaching hRPE cells and then the cells were incubated and washed to remove the unbound beads. A magnet was put to the culture plate to provide vertical magnetic force. Cytochalasin D （CD） was added to the culture fluid to inhibit the phagocytizing, secreting, and moving functions of the hRPE. Before the experiment and 15 rain, 0. 5 h, 1 h, 4 h, and 8 h the beginning of experiment,the MCP-1 mRNA and IL-8 mRNA in the hRPE cells were measured with reverse transcriptase polymerse chain reaction （RT-PCR） , and the MCP-1 and IL-8 protein expression in the supernatants was tested with enzyme-linked immunosorbent assay （ELISA） kits,Results MCP-1 and IL-8 mRNA were expressed at very low levels in the RPE cells and supematant not exposed to magnetic force. After exposure to magnetic force, both MCP-1 and IL-8 mRNA demonstrated “double peaks” expression. Their first peak levels appeared within 0.5 h, being 3.30 ng/L ± 0.12 ng/L and 1.88 ng/L ± 0.08 ng/L respectively. The first peaks of MCP-1 and IL-8 protein expression were within 1 h, being 552.05 ng/L ± 7.64 ng/Land 236.67 ng/L± 14. 30 ng/L respectively, and the second peaks appeared about 4 hours later. After cytochalasin D pretreated, the expression and secretion of both MCP-1 and IL-8 of the hRPE cells were significantly decreased to the levels of 2. 36 ng/L ± 0. 27 ng/L and 1.64 ng/L ±0. 08 ng/L,and 353.80 ng/L ± 16.68 ng/L and 101.86 ng/L ±15.92 ng/L respectively. Conclusion Mechanical stress induces the RPE cells to express MCP-1 and IL-8, and this effect was inhibited in part by pretreatment of CD

关 键 词：视网膜脱离 色素上皮 眼 视网膜疾病 单核细胞化学吸引蛋白质-1 白细胞介素-8 培养的人视网膜色素上皮细胞 MCP-1 IL-8 机械牵拉 

分 类 号：R774.1[医药卫生—眼科]