Establishment of lung microvascular endothelial cell line and activity of tyrosine kinase receptor in the cells after lipopolysaccharide treatment / 第三军医大学学报

ABSTRACT

Objective To establish a cell line of rat lung microvascular endothelial cells (LMVEC) and investigate the activity of tyrosine kinase receptor (TKR) in them after lipopolysaccharide (LPS) treatment Methods Rat LMVEC were cultured with lung tissue block pasted methods, and were identified by using immuocytochemical methods with anti human CD31 and factor Ⅷ antibody, by testing their binding with the Lectin Bandeiraea simplicifoliaⅠwith immunofluoresence staining, and by measuring the activity of angiotensin converting enzyme (ACE) with ACE kit The cells were treated with LPS at 1?g/ml for 2, 8, 16, 24 h respectively The activity of TKR in the cells was determined with radioimmunoassay Results The self cultured LMVEC in flask showed regular cobblestone morphology and positive for binding of the Lectin Bandeiraea simplicifoliaⅠand anti human CD31 antibody The TKR activity in LMVEC increased in 2 h group, reached the highest in 8 h group, and fell in 16 h group, but still significantly higher than that in normal group Conclusion TKR activity increases in the cytoplasm of LMVEC, but decreases in the cell membrane, suggesting that TKR and its correlated signal pathways may exert important modulation in the mechanism of acute lung injury and systemic inflammatory response syndrome