LPS诱导小鼠肺微血管内皮细胞VEGF表达的探讨

Researches of Mice Lung Microvessel Endothelial Cell Expressing VEGF after Lipopolysaccharide Inducing

目的:研究小鼠肺微血管内皮细胞分离培养方法并探讨脂多糖(LPS)诱导小鼠肺微血管内皮细胞血管 内皮生长因子(VEGF)表达。方法:贴块法培养小鼠肺微血管内皮细胞,并进行系列鉴定。采用ELISA方法检测 正常组、LPS(1μg/ml)作用2、8、16、24、36、48h组的VEGF变化水平。结果:获得的小鼠肺微血管内皮细胞具有鹅 卵石形态,但LPS对小鼠肺微血管内皮细胞形态影响明显。对异植物血凝素结合试验,CD3l及Ⅷ因子抗原免疫组 化染色为阳性。LPS(1μg/ml)分别作用小鼠肺微血管内皮细胞2h组可见VEGF表达升高,8h组达最高,16、24、 36、48h组的表达量低于8h组。结论:LPS作用后,小鼠肺微血管内皮细胞VEGF表达呈抛物线型下降。LPS损 伤内皮细胞功能和形态。

Objective: To establish a isolation and cultivation method of mice lung microvessel endothelial cell(LMVEC) and investigate the VEGF expression of mice LMVEC induced by lipopolysaccharide(LPS). Methods: Mice LMVEC were cultured with lung tissue block pasted method, and were identified using immunocytochemical methods with factor Ⅷ antibody, by testing their binding with the Lectin Bandeiraea simplicifolia I with immunofluorescent staining,by testing anti-human CD31 with flow cytometry. The cells were treated with LPS at 1 ug/ml for2、8、16、 24、 36、 48hours respectively, and measuring VEGF level of culture medium by ELISA. Results: The self-cultured mice LMVEC in flask showed regular cobblestone morphology but the changes in the mice LMVEC morphology was obvious after LPS stimulation,and positive for binding of the Lectin Bandeiraea simplicifolia I and anti-human CD31 and factor Ⅷ antibody.The VEGF level in mice LMVEC increased in 2 hours group, reached the highest in 8 hours group,and fell in 16、24、36、48 hours group. Conclusion: After LPS treating, the VEGF expression of mice LMVEC show the parabola descent. LPS damage the morphology and function of mice LMVEC.