Ultrasound-microbubble-mediated gene transfer of inducible Smad7 blocks transforming growth factor-beta signaling and fibrosis in rat remnant kidney.
Transforming growth factor (TGF)-beta1 has been shown to play a critical role in hypertensive nephropathy. We hypothesized that blocking TGF-beta1 signaling could attenuate renal fibrosis in a rat model of remnant kidney disease. Groups of six rats were subjected to 5/6 nephrectomy and received renal arterial injection of a doxycycline-regulated Smad7 gene or control empty vector using an ultrasound-microbubble-mediated system. Smad7 transgene expression within the kidney was tightly controlled by the addition of doxycycline in the daily drinking water. All animals were euthanized at week 4 for renal functional and histological examination. Hypertension of equivalent magnitude (190 to 200 mmHg) developed in both Smad7- and empty vector-treated rats. However, treatment with Smad7 substantially inhibited Smad2/3 activation and prevented progressive renal injury by inhibiting the rise of 24-hour proteinuria (P < 0.001) and serum creatinine (P < 0.001), preserving creatinine clearance (P < 0.05), and attenuating renal fibrosis and vascular sclerosis such as collagen I and III expression (P < 0.01) and myofibroblast accumulation (P < 0.001). In conclusion, TGF-beta/Smad signaling plays a critical role in renal fibrosis in a rat remnant kidney model. The ability of Smad7 to block Smad2/3 activation and attenuate renal and vascular sclerosis demonstrates that ultrasound-mediated Smad7 gene therapy may be a useful therapeutic strategy for the prevention of renal fibrosis in association with hypertension.
Hou
Chun-Cheng
CC
Department of Medicine, Municipal Wan-Fang Hospital, Taipei Medical University, Taipei, Taiwan.
Wang
Wansheng
W
Huang
Xiao R
XR
Fu
Ping
P
Chen
Tso-Hsiao
TH
Sheikh-Hamad
David
D
Lan
Hui Y
HY
D000818:Animals; D001794:Blood Pressure; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D004318:Doxycycline / Q000494:pharmacology; D005355:Fibrosis / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006973:Hypertension; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D007668:Kidney / Q000378:metabolism* / Q000473:pathology; D008297:Male; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D051906:Smad7 Protein; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D019076:Transgenes; D014465:Ultrasonics
D004268:DNA-Binding Proteins; D051906:Smad7 Protein; C497808:Smad7 protein, rat; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D004318:Doxycycline
J Clin Invest. 1999 Jul;104(1):5-11
10393693
J Hypertens. 1998 Nov;16(11):1603-9
9856360
Mol Cell. 2000 Dec;6(6):1365-75
11163210
J Biol Chem. 2001 Mar 9;276(10):6983-92
11114293
Contrib Nephrol. 2001;(135):153-60
11705280
Kidney Int. 2002 Jan;61(1 Suppl):S94-8
11841620
Am J Kidney Dis. 2002 Mar;39(3):486-92
11877567
J Am Soc Nephrol. 2002 Apr;13(4):887-93
11912247
J Am Soc Nephrol. 2002 Apr;13(4):1126-8
11912276
Diabetes Care. 2002 Jun;25(6):1072-7
12032117
J Am Soc Nephrol. 2002 Jun;13(6):1464-72
12039975
Biochem Biophys Res Commun. 2002 Sep 6;296(5):1356-65
12207925
J Am Soc Nephrol. 2002 Oct;13(10):2600-10
12239251
J Am Soc Nephrol. 2002 Nov;13(11):2657-66
12397035
Am J Kidney Dis. 2003 Mar;41(3 Suppl 1):S3-7
12612942
Kidney Int. 2003 Jun;63(6):2010-9
12753288
J Am Soc Nephrol. 2003 Jun;14(6):1535-48
12761254
J Am Soc Nephrol. 2003 Jul;14(7):1816-24
12819241
J Am Soc Nephrol. 2003 Jul;14(7 Suppl 2):S92-8
12819310
Gastroenterology. 2003 Jul;125(1):178-91
12851882
J Am Soc Nephrol. 2004 Jan;15 Suppl 1:S55-7
14684674
FASEB J. 2004 Jan;18(1):176-8
12709399
Am J Pathol. 2004 Apr;164(4):1389-97
15039226
Am J Pathol. 1992 Jan;140(1):95-107
1731533
J Clin Invest. 1992 Aug;90(2):456-61
1644917
J Clin Invest. 1993 Dec;92(6):2597-601
8254017
J Clin Invest. 1994 Jun;93(6):2431-7
8200978
Kidney Int. 2000 May;57(5):1803-17
10792600
Nat Med. 1996 Apr;2(4):418-23
8597951
Nephrol Dial Transplant. 1996 Feb;11(2):263-9
8671777
Kidney Int. 1996 Jul;50(1):148-55
8807583
Hypertension. 1998 Jan;31(1 Pt 2):181-8
9453300
Miner Electrolyte Metab. 1998;24(2-3):174-80
9525702
Curr Opin Genet Dev. 1998 Feb;8(1):103-11
9529613
Kidney Int. 1998 Sep;54(3):864-76
9734611
Kidney Int. 1998 Nov;54(5):1570-80
9844133
J Mol Med (Berl). 1999 Jul;77(7):556-64
10494801