DNAzyme for TGF-beta suppressed extracellular matrix accumulation in experimental glomerulonephritis.

Abstract:

BACKGROUND:We developed an electroporation-mediated gene transfer method targeting glomerular mesangial cells. Injecting DNA solution via renal artery followed by electric pulses using tweezers-type electrodes could result in efficient transfection in mesangial cells. Therefore, this gene transfer system opened a feasible strategy to manipulate the function of several cytokines and growth factors in mesangial cells. Recently, a new generation of catalytic nucleic acid composed of DNA, named DNA enzyme (DNAzyme), has been developed. METHOD:We generated a DNAzyme (TGFDE) targeting transforming growth factor-beta1 (TGF-beta1), and examined the therapeutic effect of TGFDE in vitro and in vivo. RESULTS:In cultured rat mesangial cells, treatment with TGFDE blocked TGF-beta1 mRNA expression, and thereby suppressed type I collagen mRNA expression. Next, we introduced TGFDE or scrambled DNAzyme (TGFSCR) into anti-Thy-1 model of nephritic rats by electroporation 3 days after disease induction. Northern blot analysis and immunohistochemical staining demonstrated that glomerular message and protein expression of TGF-beta1, alpha-smooth muscle actin (alpha-SMA), and type I collagen were suppressed in TGFDE-transfected nephritic rats compared with untreated nephritic rats and TGFSCR-transfected rats on day 7. Consequently, we observed significant reduction in glomerular matrix score in TGFDE-transfected nephritic rats. CONCLUSION:Inhibition of TGF-beta1 expression by electroporation-mediated DNAzyme transfer might be useful for the therapy of glomerulonephritis.

journal_name

Kidney Int

journal_title

Kidney international

authors

Isaka Y,Nakamura H,Mizui M,Takabatake Y,Horio M,Kawachi H,Shimizu F,Imai E,Hori M

doi

10.1111/j.1523-1755.2004.00777.x

subject

Has Abstract

pub_date

2004-08-01 00:00:00

pages

586-90

issue

2

eissn

0085-2538

issn

1523-1755

pii

S0085-2538(15)50085-9

journal_volume

66

pub_type

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