Up-regulation of parathyroid hormone-related protein in folic acid-induced acute renal failure.
BACKGROUND
Parathyroid hormone (PTH)-related protein (PTHrP) is present in many normal tissues, including the kidney. Current evidence supports that PTHrP is involved in renal pathophysiology, although its role on the mechanisms of renal damage and/or repair is unclear. Our present study examined the changes in PTHrP and the PTH/PTHrP receptor (type 1) in folic acid-induced acute renal failure in rats. The possible role of PTHrP on the process of renal regeneration following folic acid administration, and potential interaction between angiotensin II (Ang II) and endothelin-1, and PTHrP, were examined in this animal model.


METHODS
PTHrP, PTH/PTHrP receptor, ACE, and preproendothelin-1 (preproET-1) mRNA levels in the rat kidney were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and/or RNase protection assay. Immunohistochemistry also was performed for PTHrP, the PTH/PTHrP receptor, and Ang II in the renal tissue of folic acid-injected rats. The role of PTHrP on tubular cell proliferation following folic acid injury was investigated in vitro in rat renal epithelial cells (NRK 52E). PTHrP secretion in the medium conditioned by these cells was measured by an immunoradiometric assay specific for the 1-36 sequence.


RESULTS
Using RT-PCR, PTHrP mRNA was rapidly (1 hour) and maximally increased (3-fold) in the rat kidney after folic acid, decreasing after six hours. At 72 hours, renal function was maximally decreased in these rats, associated with an increased PTHrP immunostaining in both renal tubules and glomeruli. In contrast, the PTH/PTHrP receptor mRNA (RNase protection assay) decreased shortly after folic acid administration. Moreover, PTH/PTHrP receptor immunostaining dramatically decreased in renal tubular cell membranes after folic acid. A single subcutaneous administration of PTHrP (1-36), 3 or 50 microg/kg body weight, shortly after folic acid injection increased the number of tubular cells staining for proliferating cell nuclear antigen by 30% (P < 0.05) or 50% (P < 0.01), respectively, in these rats at 24 hours, without significant changes in either renal function or calcemia. On the other hand, this peptide failed to modify the increase (2-fold over control) in ACE mRNA, associated with a prominent Ang II staining into tubular cell nuclei, in the kidney of folic acid-treated rats at this time period. The addition of 10 mmol/L folic acid to NRK 52E cells caused a twofold increase in PTHrP mRNA at six hours, without significant changes in the PTH/PTHrP receptor mRNA. The presence of two anti-PTHrP antibodies, with or without folic acid, in the cell-conditioned medium decreased (40%, P < 0.01) cell growth.


CONCLUSIONS
Renal PTHrP was rapidly and transiently increased in rats with folic acid-induced acute renal failure, featuring as an early response gene. In addition, changes in ACE and Ang II expression were also found in these animals. PTHrP induces a mitogenic response in folic acid-damaged renal tubular cells both in vivo and in vitro. Our results support the notion that PTHrP up-regulation participates in the regenerative process in this model of acute renal failure and is a common event associated with the mechanisms of renal injury and repair.
Santos
S
S
Bone and Mineral Metabolism Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Bosch
R J
RJ
Ortega
A
A
Largo
R
R
Fernández-Agulló
T
T
Gazapo
R
R
Egido
J
J
Esbrit
P
P
D058186:Acute Kidney Injury / Q000139:chemically induced / Q000378:metabolism*; D000804:Angiotensin II / Q000032:analysis; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D004195:Disease Models, Animal; D019332:Endothelin-1; D016232:Endothelins / Q000032:analysis; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005492:Folic Acid; D007150:Immunohistochemistry; D007672:Kidney Cortex / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D007684:Kidney Tubules / Q000187:drug effects / Q000473:pathology; D008297:Male; D044162:Parathyroid Hormone-Related Protein; D007703:Peptidyl-Dipeptidase A / Q000032:analysis; D011498:Protein Precursors / Q000032:analysis; D011506:Proteins / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012038:Regeneration; D015854:Up-Regulation
D019332:Endothelin-1; D016232:Endothelins; D044162:Parathyroid Hormone-Related Protein; D011498:Protein Precursors; D011506:Proteins; D000804:Angiotensin II; D005492:Folic Acid; D007703:Peptidyl-Dipeptidase A