Antibody to transforming growth factor-beta ameliorates tubular apoptosis in unilateral ureteral obstruction.
BACKGROUND
Unilateral ureteral obstruction (UUO) is characterized by progressive renal atrophy, renal interstitial fibrosis, an increase in renal transforming growth factor-beta (TGF-beta), and renal tubular apoptosis. The present study was undertaken to determine the effect of a monoclonal antibody to TGF-beta (1D11) in UUO.


METHODS
Mechanical stretch was applied to tubular epithelial cells (NRK-52E) by a computer-assisted system. Three doses of 1D11 (either 0.5, 2, or 4 mg/rat) were administered to rats one day prior to UUO and every two days thereafter, and kidneys were harvested at day 13. Fibrosis was assessed by measuring tissue hydroxyproline and mRNA for collagen and fibronectin. Apoptosis was assessed with the terminal deoxy transferase uridine triphosphate nick end-labeling assay. TGF-beta levels were determined by bioassay. Western blot and immunostaining were used to identify proliferating cell nuclear antigen (PCNA), p53, bcl-2, and inducible nitric oxide synthase (iNOS).


RESULTS
Stretch significantly induced apoptosis in NRK-52E cells, which was accompanied by an increased release of TGF-beta; 1D11 (10 microg/mL) totally inhibited stretch-induced apoptosis. Control obstructed kidney contained 20-fold higher TGF-beta as compared with its unobstructed kidney; 1D11 neutralized tissue TGF-beta of the obstructed kidney. Control obstructed kidney exhibited significantly more fibrosis and tubular apoptosis than its unobstructed counterpart, which was blunted by 1D11. In contrast, 1D11 significantly increased tubular proliferation. p53 immunostaining was localized to renal tubular nuclei of control obstructed kidney and was diminished by 1D11. In contrast, bcl-2 was up-regulated in the 1D11-treated obstructed kidney. Total NOS activity and iNOS activity of the obstructed kidney were increased by 1D11 treatment.


CONCLUSIONS
The present study strongly suggests that an antibody to TGF-beta is a promising agent to prevent renal tubular fibrosis and apoptosis in UUO.
Miyajima
A
A
Center for Pediatric Urology and Laboratory for Minimally Invasive Urologic Surgery, New York, New York, USA.
Chen
J
J
Lawrence
C
C
Ledbetter
S
S
Soslow
R A
RA
Stern
J
J
Jha
S
S
Pigato
J
J
Lemer
M L
ML
Poppas
D P
DP
Vaughan
E D
ED
Felsen
D
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis; D001284:Atrophy; D015153:Blotting, Western; D002460:Cell Line; D002956:Citrulline; D005355:Fibrosis; D015971:Gene Expression Regulation, Enzymologic; D020287:In Situ Nick-End Labeling; D007684:Kidney Tubules / Q000737:chemistry / Q000201:enzymology / Q000473:pathology*; D009395:Nephritis, Interstitial / Q000473:pathology / Q000517:prevention & control / Q000628:therapy; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013314:Stress, Mechanical; D016212:Transforming Growth Factor beta / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000032:analysis; D014517:Ureteral Obstruction / Q000473:pathology* / Q000517:prevention & control / Q000628:therapy*
D000911:Antibodies, Monoclonal; D018809:Proliferating Cell Nuclear Antigen; D019253:Proto-Oncogene Proteins c-bcl-2; D016212:Transforming Growth Factor beta; D016159:Tumor Suppressor Protein p53; D002956:Citrulline; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat