| 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].现代生物医学进展英文版,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 |
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| 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） |
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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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