文章摘要
叶巍 夏风杰 姚瑶 曹谦 闫峰 黄振平.机械牵拉诱导人RPE细胞IL-8分泌及其对PI3K/AKT 通路的影响[J].,2016,16(22):4217-4220
机械牵拉诱导人RPE细胞IL-8分泌及其对PI3K/AKT 通路的影响
IL-8 Secretion Induced by Mechanical Stretch in HumanRPE Cells and Its Effect on PI3K/AKT Pathway
  
DOI:
中文关键词: 机械应力  视网膜色素上皮细胞  PI3K/AKT 通路  上皮-间质转化
英文关键词: Mechanical stress  Retinal pigment epitheliumcell  PI3K/AKT pathway  Epithelial mesenchymal transition
基金项目:国家自然科学基金项目(81200719)
作者单位
叶巍 夏风杰 姚瑶 曹谦 闫峰 黄振平 第二军医大学附属南京军区总医院眼科南京大学医学院附属金陵医院 
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中文摘要:
      目的:视网膜脱离发生机制尚不明确,本文旨在研究机械应力牵拉后视网膜色素上皮细胞上清中IL-8 分泌量的变化以及人 其对RPE 细胞PI3K/AKT 通路变化影响。方法:培养人RPE 细胞,应用Flex-cell细胞应力加载系统牵拉人RPE 细胞不同时间(0 h、1 h、3 h、6 h、9 h)建立不同牵拉时间的模型。分别标记为对照组、牵拉1 h组、牵拉3 h组、牵拉6 h组、牵拉9 h 组。随后用 ELISA 方法检测人RPE 细胞不同实验组上清中IL-8 分泌量的变化。细胞免疫荧光和Western blot 方法观察PI3K、PPI3K、 T-AKT、P-AKT 的表达。结果:随着牵拉时间的延长,ELISA 方法检测人RPE 细胞不同实验组上清中IL-8 分泌量逐渐增加 [924.79± 5.92 pg/mL、947.73± 5.34 pg/mL、974.53± 5.74 pg/mL、979.57± 1.12 pg/mL、1019.22± 4.25 pg/mL],差异有统计学意义 (F=166,p<0.01),各牵拉组与对照组分别对比,差异有统计学意义(P<0.05)。细胞中P-AKT 蛋白表达增加[0.61± 0.02、0.97± 0.05、 0.99± 0.04、1.21± 0.11、1.20± 0.07],差异有统计学意义(F=41.24,p<0.01),各牵拉组与对照组分别对比,差异有统计学意义(P<0. 05),AKT 表达未见明显变化。结论:机械应力牵拉人RPE细胞时,细胞上清中IL-8 分泌量逐渐增加同时RPE 细胞内PI3K/AKT 通路明显激活,并与牵拉时间有关,为探究EMT 原因和对防治PVR提供了理论基础。
英文摘要:
      Objective:The mechanism of retinal detachment is not clear, this paper aims to study volume changes of supernatant IL-8 and the changes of of PI3K / Akt pathway in human RPE cells which were stretched by the Device of Mechanical Stress.Methods:Human RPE cells were stretched by the Device of Mechanical Stress for 0 h,1 h,3 h,6 h,9 h. Labeled as the control group, 1 h group, 3 h group, 6 h group, 9 h group. The changes of IL-8 secreted by human RPE cells in different experimental groups were detected by ELISA . Expression levels of PPI3K, T-AKT, PI3K P-AKT were observed by the immunofluorescence and Western blot method.Results:With the extension of stretch time, the volume of supernatant IL-8 in different groups increased gradually [924.79± 5.92 pg/mL, 947.73± 5.34 pg/mL, 974.53± 5.74 pg/mL, 979.57± 1.12 pg/mL, 1019.22± 4.25pg/mL]. The results reviewed that the differences between different groups were significant (F=166, P<0.01). The differences between stretch groups and control group were significant (P < 0.05). The protein expression of PAKT increased [0.61± 0.02, 0.97± 0.05, 0.99± 0.04, 1.21± 0.11, 1.20± 0.07]. The differences between different groups were significant (F=41.24, P < 0.01). The differences between stretch groups and control group were significant (P < 0.05).Conclusion:With the extension of stretch time, the volumes of supernatant IL-8 in different groups increased gradually and the PI3K / Akt pathway was activated in human RPE cells which was stretched by the Device of Mechanical Stress. The results offer a method to explore the reasons of EMT and provide a basis for the prevention and treatment of PVR.
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