蛋白酪氨酸激酶抑制剂对RPE细胞趋化因子表达和Ca^(2+)的影响  

Effects of protein tyrosine kinase inhibitor on expression of chemotatic factors and Ca^(2+) in RPE cells

作　　者：张世杰[1] 张皙[2] 

机构地区：[1]北京大学第一医院,北京市100034 [2]上海市第一人民医院,上海市200080

出　　处：《眼科新进展》2010年第9期828-831,共4页Recent Advances in Ophthalmology

摘　　要：目的研究蛋白酪氨酸激酶(protein tyrosine kinase,PTK)抑制剂染料木黄酮(Genistein)对白介素-1β(interleukin-1β,IL-1β)诱导的视网膜色素上皮(retinal pigment epithelium,RPE)细胞趋化因子表达和细胞内Ca2+的影响。方法应用不同剂量IL-1β对RPE细胞进行刺激,RT-PCR法观察细胞IL-8和单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)mRNA表达的变化。应用Genistein对RPE细胞培养进行干预,观察其对IL-1β诱导RPE细胞IL-8和MCP-1 mRNA表达的影响。应用Fura2/AM、荧光分光光度计检测IL-1β和Genistein对RPE细胞内Ca2+的影响。结果 RT-PCR结果显示,0.1μg.L-1、1μg.L-1、10μg.L-1IL-1β刺激组IL-8 mRNA相对表达值分别为0.446±0.133、0.727±0.054和0.523±0.106,对照组为0.272±0.088;1μg.L-1、10μg.L-1IL-1β刺激组与对照组比较,差异有统计学意义(P=0.001、0.015)。10μg.L-1IL-1β诱导的RPE细胞MCP-1 mRNA相对表达值与对照组比较,差异有显著统计学意义(P=0.002)。10mg.L-1、50mg·L-1 Genistein对IL-1β诱导的IL-8和MCP-1 mRNA表达具有明显的抑制作用,差异均有显著统计学意义(均为P<0.01)。IL-1β引起细胞内Ca2+浓度升高,Genistein对细胞内Ca2+作用与之相反。结论 Genistein对IL-1β诱导的IL-8和MCP-1 mRNA表达具有明显的抑制作用,其机制可能为抑制IL-1β诱导的细胞内Ca2+增加。Objective To investigate the effects of protein tyrosine kinase inhibitor-Genistein on expression of interleukin-1β（IL-1β） induced chemotatic factors and intracellular Ca^2＋ in retinal pigment epithelium（RPE） cells.Methods Different doses of IL-1β were used to stimulate RPE cells.RT-PCR method was used to observe the changes of IL-8 and monocyte chemotactic protein-1（MCP-1） mRNA.Genistein was used to interfere RPE cell culturing,and observed its effects on expression of IL-1β induced IL-8 and MCP-1 mRNA in RPE cells.Fura2/AM fluorescent probe and spectrofluorometer were used to detect the effects of IL-1β and Genistein on intracellular Ca2＋ in RPE cells.Results RT-PCR results indicated that the relative expression of IL-8 mRNA were 0.446±0.133,0.727±0.054 and 0.523±0.106 in 0.1 μg·L-1,1 μg·L-1 and 10 μg·L-1 IL-1β stimulated groups,0.272±0.088 in control group,and there were statistical differences for 1 μg·L-1 and 10 μg·L-1 IL-1β group compared with control group（P=0.001,0.015）.There was significant difference in relative expression of MCP-1 mRNA between 10 μg·L-1 IL-1β stimulated group and control group（P=0.002）.The 10 mg·L-1 and 50 mg·L-1 Genistein had obviously inhibitory effect on expression of IL-8 and MCP-1 mRNA induced by IL-1β,and there were significant differences（all P0.01）.IL-1β could increase the intracellular Ca^2＋ levels,while Genistein had opposite effect on intracellular Ca^2＋.Conclusion Genistein has obviously inhibitory effect on expression of IL-8 and MCP-1 mRNA induced by IL-1β,which may be through inhibiting the rising of intracellular Ca^2＋ induced by IL-1β.

关 键 词：视网膜色素上皮细胞 趋化因子 蛋白酪氨酸激酶抑制剂 白介素-1Β 细胞内CA2+ 

分 类 号：R774.1[医药卫生—眼科]