[Effect of plasminogen activator inhibitor 1 gene transfer into mesangial cells on Extracellular matrix accumulation]
1Department of Nephrology, General Hospital of PLA, Kidney Center of PLA, Beijing 100853.
Objective:
To observe the influence of plasminogen activator inhibitor 1 (PAI-1) overexpression on extracellular matrix (ECM) accumulation of rat mesangial cells.
Methods:
PAI-1/GFP green fluorescent protein fusion gene eukaryotic expression vector pCMX-PAI-1-GFP was constructed and delivered into cultured rat mesangial cells by liposome. PAI-1/GFP expression was observed by fluorescent microscope. FN, LN and Type IV collagen levels were measured by Northern blot analysis and ELISA method.
Results:
Northern blot analysis, SDS-PAGE analysis and Western blot analysis demonstrated that PAI-1/GFP gene can be transcripted and translated in rat mesangial cells. PAI-1/GFP gene can express a specific protein. The recombinant PAI-1/GFP had significant inhibition activity on urokinase-type plasminogen activator and displayed autonomous fluorescence. The PA activity of cell supernatants decreased markedly in gene transfer group. FN, LN and Type IV collagen levels were higher than in a control group.
Conclusion:
Over-expression of PAI-1 gene leads to mesangial ECM accumulation. The changes of PA/PAI-1 activity in supernatants of cultured rat mesangial cells might be the key factor in ECM accumulation regulation.
