SP600125对周期牵张致A549细胞上调白细胞介素-8表达的影响  

The effects of SP600125 on the upregulation of IL-8 expression in A549 cell induced by cyclic stretching

作　　者：王月兰[1] 刘玉华[2] 姚尚龙[3] 

机构地区：[1]山东大学千佛山医院麻醉科,济南市250014 [2]河北省医学情报研究所 [3]华中科技大学同济医学院协和医院麻醉科

出　　处：《中华麻醉学杂志》2006年第7期646-648,共3页Chinese Journal of Anesthesiology

基　　金：教育部博士点基金资助项目(20020487063);国家自然基金资助项目(30471661)

摘　　要：目的探讨SP600125对周期牵张致A549细胞上调白细胞介素-8(IL-8)表达的影响。方法将A549细胞以密度为5×105种植于特制的硅胶树脂膜上,DMEM培养基培养48 h后,置于细胞牵张仪行周期性牵张。牵张组:牵张强度为细胞体积扩大15％,牵张周期:30个周期／min,牵张时间分别为0(未进行牵张)、15、30、60、120 min。预防处理组:将JNK抑制剂SP600125加入培养基,终浓度为50μmmol／L孵育30 min后再行上述处理,收集细胞和培养液,提取细胞中的总蛋白和RNA,用RT-PCR方法测定细胞IL-8 mRNA表达,用Western blot测定细胞JNK和磷酸化JNK(p-JNK)的水平,采用ELISA法测定细胞培养液中IL-8浓度。结果周期牵张30～120 min可导致牵张组A549细胞IL-8 mRNA、IL- 8浓度增加,且呈时间依赖性,周期牵张120 min时达高峰,p-JNK水平升高;SP600125可减弱周期牵张导致的上述改变(P<0．05)。结论周期牵张致A549细胞IL-8的上调与JNK的激活有关。Objective To investigate the effects of SP600125 on the upregulation of interleukin-8 （IL-8） expression in A549 cells induced by cyclic stretching.Methods A549 cells of human lung adenocarcinoma cell line were cultured in DMEM culture medium for 48 h, and were inoculated on fibronectin-coated silicone clastomeric membrane at the density of 5 x 105 and incubated for another 48h. The cells were then divided into 2 groups： Ⅰ stretch group and Ⅱ SP600125 pretreatment ＋ stretch group. The cells were stretched and enlarged by 15% at 30 cycles per minute for 0, 15, 30, 60 and 120 min. In group Ⅱ SP600125 50 μmol·L^-1 was added to culture medium 30 min before the cells were stretched. The total protein and RNA in the cells were obtained after being stretched for determination of IL-8 mRNA expression （ by RT-PCR） and phospho-JNK （p-JNK） activity （Western blot）. The concentration of IL-8 in the culture medium was determined （by EIdSA）.Results Cyclic stretching significantly increased IL-8 mRNA expression and p-JNK activity in A549 cells and IL-8 concentration in culture medium time-dependently in group Ⅰ and the peak was reached at 120 min. In group Ⅱ pretreatment with SP600125 inhibited the changes induced by cyclic stretching. Conclusion Upregulation of IL-8 expression in A549 ceils induced by cyclic stretching is closely associated with the activation of JNK.

关 键 词：细胞系 肿瘤 白细胞介素8 丝裂原激活蛋白激酶类 周期牵张 

分 类 号：R734.2[医药卫生—肿瘤]