Ex vivo transfer of the decorin gene into rat glomerulus via a mesangial cell vector suppressed extracellular matrix accumulation in experimental glomerulonephritis.
The activation of transforming growth factor-beta (TGF-beta) is known to be one of the major causes of glomerulosclerosis. Decorin (DCN) is a natural inhibitor of TGF. The purpose of this study was to assess the feasibility of transferring the DCN gene to antithymocyte serum (ATS) glomerulonephritis glomeruli via a mesangial cell vector to treat glomerulonephritis fibrosis. For this process, the recombinant eukaryotic expression plasmid pcDNA3.1A-DCN was constructed and transfected into mesangial cell. The DCN-positive cloned cells were transferred to rat antithymocyte serum glomeruli by a left renal artery injection. Using immunohistochemical staining, approximately 37-60% (48.6% +/- 11.34%; mean +/- SE, n = 8) of the glomeruli were BrdU-positive in the injected-side kidney. DCN proteins were observed in the cytoplast beginning 12 h after injection. TGF-beta1 expression in the injected side glomeruli decreased significantly at day 4 (P < 0.05), compared with that in the uninjected-side kidney. The expression leaves of fibronectin and collagen IV decreased significantly at days 1-2 (P < 0.01) and day 4 (fibronectin, P < 0.01; collagen IV, P < 0.05). These results suggest that the use of DCN can decrease antithymocyte serum glomerulonephritis extracellular matrix (ECM) ingredients and that such use offers a favorable experimental basis for gene therapy for kidney disease.
Huijun
Wang
W
Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA. huijuwan@mdanderson.org
Long
Chen
C
Zhigang
Zhang
Z
Feng
Jiang
J
Muyi
Guo
G
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000235:genetics / Q000378:metabolism; D058575:Decorin; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D005822:Genetic Vectors; D005920:Glomerular Mesangium / Q000378:metabolism* / Q000473:pathology; D015432:Glomerulonephritis, Membranoproliferative / Q000378:metabolism*; D007106:Immune Sera / Q000276:immunology; D007124:Immunoenzyme Techniques; D007678:Kidney Glomerulus / Q000378:metabolism* / Q000473:pathology; D008297:Male; D011509:Proteoglycans / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018800:Thy-1 Antigens / Q000276:immunology; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors*
D024141:Collagen Type IV; C546880:Dcn protein, rat; D058575:Decorin; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D007106:Immune Sera; D011509:Proteoglycans; D018800:Thy-1 Antigens; D016212:Transforming Growth Factor beta; D001973:Bromodeoxyuridine