EphA2在内毒素诱导人肺血管内皮细胞凋亡模型中表达与意义的研究

The expression and the role of EphA2 in lipopolysaccharides induced apoptosis model of human pulmonary microvascular endothelial cells

目的探讨EphA2在内毒素（lipopolysaccharides，LPS）所致人肺血管内皮细胞（human pulmonary microvascular endothelial cells，HPMVECs）凋亡模型中的表达与作用。方法体外培养HPMVECs（24瓶），采用完全随机分组法分为4组（每组6瓶）：对照组（C组）、EphA2抑制剂组（Fc组）、LPS干预组（LPS组）和LPS＋EphA2抑制剂组（LPS＋Fc组）。流式细胞仪Annexin V／PI法检测每组细胞凋亡率，酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测各组细胞上清液中肿瘤坏死因子（tumor necrosis factor，TNF）-α水平，Western blot检测各组EphA2水平。结果与C组比较，LPS组细胞的凋亡率升高[（7．8±1．4）％，（23．1±2．9）％]（P〈0．05），TNF啾的量显著增加[（12．4±2．6），（37．1±5．3）ng/L]（P〈0．05），EphA2蛋白表达水平明显升高[（1．118±0．093），（2．244±0．127）]（P〈0．05）；与LPS组比较，LPS＋Fc组细胞凋亡率下降[（23．1±2．9）％，（15．1±1．5）％]（P〈0．05），TNF-α的量显著降低[（37．1±5．3），（25．5±2．9）ng/L]（P〈0．05），EphA2表达明显下降[（2．244±0．127），（1．502±0．063）]（P〈0．05）；而C组与Fc组比较，细胞凋亡率[（7．8±1．4）％，（7．4±1．3）％]、TNF-α的量[（12．4±2．6），（14．2±2．3）ng／L]、EphA2表达量[（1．118±0．093），0．943±0．014）]差异无统计学意义（P〉0．05）。结论LPS诱导的HPMVECs炎性反应凋亡模型中，抑制EphA2的表达可以减轻细胞凋亡。

Objective To investigate the expression and the role of EphA2 in lipopolysaccharides （LPS） induced apoptosis model of human pulmonary microvascular endothelial cells （HPMVECs）. Methods HPMVECs were cultivated in vitro, and were randomly divided into four groups： control group （group C）, Fc group, LPS group, LPS＋Fc group. Apoptosis was detected by flow cytometry Annexin-FITC and PI double staining, western blot was performed to detect protein EphA2 expression in human pulmonary microvascular endothelial cells, and the concentration of tumor necrosis factor （TNF）-α was determined by enzyme-linked immunosorbent assay（ELISA）. Results Compared with group C, apoptosis rate was significantly inereased[（7.8±1.4）%,（23.1±2.9）%] in LPS group （P〈0.05）. TNF-α concentration in LPS group was increased compared with group 12 [ （12.4±2.6）, （37.1±5.3） ng/L]（P〈0.05 ）. EphA2 protein expression of LPS group was increased [ （1.118±0.093 ）, （2.244±0.127）] compared with group 12 （P〈0.05）. Compared with LPS group, apoptosis rate was significantly decreased [ （ 23.1 ±2.9 ）%, （15.1±1.5 ） % ] in LPS＋Fc group （P〈0.05）, TNF-α concentration was significantly decreased in LPS＋Fc group [ （37.1±5.3）, （25.5±2.9） ng/L] （P〈0.05）, and EphA2 protein expression in LPS＋Fc group was significantly decreased [ （ 2.244±0.127 ）, （ 1.502±0.063 ） ] （P〈0.05）. Compared with group 12, no statistically difference was found in apoptosis rate[ （ 7.8± 1.4 ）%, （7.4± 1.3 ）% ] （P〉0.05）, in TNF -α concertration[ （ 12.4±2.6 ）, （ 14.2±2.3 ） ng/L ] （P〉0.05）, in EphA2 protein expression [（1.118±0.093）, （0.943±0.014）]（P〉0.05）of Fc group. Conclusions In LPS-induced HPMVE12s lung inflammation and apoptosis models, inhibiting the expression of EphA2 can reduce LPS-induced cell apoptposis.