Chimeric DNA-RNA hammerhead ribozyme targeting transforming growth factor-beta1 mRNA ameliorates renal injury in hypertensive rats.
OBJECTIVE
Transforming growth factor (TGF)-beta is a critical factor in the progression of renal injury, regardless of the primary etiology. Such injury is characterized by glomerular sclerosis and tubulointerstitial fibrosis. To develop a ribozyme-based therapy for progressive renal diseases, we examined the effects of chimeric DNA-RNA hammerhead ribozyme targeting TGF-beta1 mRNA on glomerulosclerosis in salt-loaded, stroke-prone spontaneously hypertensive rats (SHR-SP) and salt-sensitive Dahl (Dahl-S) rats.


METHODS
The chimeric DNA-RNA ribozyme to TGF-beta1 was delivered by polyethylenimine to cultured mesangial cells from SHR-SP in vitro and to glomeruli in SHR-SP in vivo. The chimeric ribozyme reduced expression of TGF-beta1 mRNA and protein, which was accompanied by inhibition of expression of extracellular matrix molecules such as fibronectin and collagen type I in mesangial cells from SHR-SP in vitro.


RESULTS
One intraperitoneal injection of 200 microg of chimeric DNA-RNA ribozyme to TGF-beta1 in vivo markedly ameliorated thickening of capillary artery walls and glomerulosclerosis in salt-loaded SHR-SP and Dahl-S rats without a reduction in blood pressure. The chimeric ribozyme reduced expression of TGF-beta1 and connective tissue growth factor (CTGF) mRNAs in renal cortex in salt-loaded Dahl-S rats. Chimeric ribozyme to TGF-beta1 significantly reduced levels of protein in urine in the Dahl-S rats.


CONCLUSIONS
These results suggest that chimeric DNA-RNA ribozyme to TGF-beta1 may be useful as a gene therapy for progressive tissue injury in a wide variety of renal diseases, including hypertensive nephrosclerosis.
Tahira
Yoshiko
Y
Department of Medicine, Nihon University School of Medicine, Division of Nephrology and Endocrinology, Tokyo, Japan.
Fukuda
Noboru
N
Endo
Morito
M
Ueno
Takahiro
T
Matsuda
Hiroyuki
H
Saito
Satoshi
S
Matsumoto
Koichi
K
Mugishima
Hideo
H
D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D005786:Gene Expression Regulation / Q000187:drug effects*; D006651:Histocytochemistry; D007678:Kidney Glomerulus / Q000187:drug effects / Q000473:pathology; D008297:Male; D009400:Nephrosclerosis / Q000188:drug therapy*; D016337:RNA, Catalytic / Q000096:biosynthesis / Q000494:pharmacology*; D012333:RNA, Messenger / Q000187:drug effects*; D051381:Rats; D020303:Rats, Inbred Dahl; D011918:Rats, Inbred SHR; D012598:Sclerosis / Q000188:drug therapy; D012720:Severity of Illness Index; D017673:Sodium Chloride, Dietary / Q000009:adverse effects; D053773:Transforming Growth Factor beta1 / Q000235:genetics* / Q000378:metabolism
D016337:RNA, Catalytic; D012333:RNA, Messenger; D017673:Sodium Chloride, Dietary; D053773:Transforming Growth Factor beta1; C427204:hammerhead ribozyme