[Effect of plasminogen activator inhibitor 1 gene transfer into mesangial cells on Extracellular matrix accumulation].
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.


CONCLUSIONS
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.
Yu
Z
Z
Department of Nephrology, General Hospital of PLA, Kidney Center of PLA, Beijing 100853.
Chen
X
X
Liao
H
H
Ye
Y
Y
Fu
B
B
D000818:Animals; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005920:Glomerular Mesangium / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis / Q000235:genetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D016326:Extracellular Matrix Proteins; D008164:Luminescent Proteins; D017395:Plasminogen Activator Inhibitor 1; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins