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Effect of variations in dietary Pi intake on intestinal Pi transporters (NaPi-IIb, PiT-1, and PiT-2) and phosphate-regulating factors (PTH, FGF-23, and MEPE)

  • Tatiana Martins Aniteli
  • Flávia Ramos de Siqueira
  • Luciene Machado dos Reis
  • Wagner Vasques Dominguez
  • Elizabeth Maria Costa de Oliveira
  • Patrícia Castelucci
  • Rosa Maria Affonso Moysés
  • Vanda JorgettiEmail author
Ion channels, receptors and transporters
Part of the following topical collections:
  1. Ion channels, receptors and transporters

Abstract

Hyperphosphatemia is a common condition in patients with chronic kidney disease (CKD) and can lead to bone disease, vascular calcification, and increased risks of cardiovascular disease and mortality. Inorganic phosphate (Pi) is absorbed in the intestine, an important step in the maintenance of homeostasis. In CKD, it is not clear to what extent Pi absorption is modulated by dietary Pi. Thus, we investigated 5/6 nephrectomized (Nx) Wistar rats to test whether acute variations in dietary Pi concentration over 2 days would alter hormones involved in Pi metabolism, expression of sodium-phosphate cotransporters, apoptosis, and the expression of matrix extracellular phosphoglycoprotein (MEPE) in different segments of the small intestine. The animals were divided into groups receiving different levels of dietary phosphate: low (Nx/LPi), normal (Nx/NPi), and high (Nx/HPi). Serum phosphate, fractional excretion of phosphate, intact serum fibroblast growth factor 23 (FGF-23), and parathyroid hormone (PTH) were significantly higher and ionized calcium was significantly lower in the Nx/HPi group than in the Nx/LPi group. The expression levels of NaPi-IIb and PiT-1/2 were increased in the total jejunum mucosa of the Nx/LPi group compared with the Nx/HPi group. Modification of Pi concentration in the diet affected the apoptosis of enterocytes, particularly with Pi overload. MEPE expression was higher in the Nx/HPi group than in the Nx/NPi. These data reveal the importance of early control of Pi in uremia to prevent an increase in serum PTH and FGF-23. Uremia may be a determining factor that explains the expressional modulation of the cotransporters in the small intestine segments.

Keywords

Intestinal absorption Sodium-phosphate cotransporters Apoptosis MEPE Uremia 

Abbreviations

Pi

Inorganic phosphate

mRNA

Messenger ribonucleic acid

FPe

Fractional excretion of phosphate

PTH

Parathyroid hormone

FGF-23

Fibroblast growth factor 23

PCR

Polymerase chain reaction

RT-qPCR

Reverse transcriptase-quantitative PCR

PBS

Phosphate-buffered saline

TBS

Tris-buffered saline

Notes

Acknowledgements

The authors thank Flávia Gomes Machado and Walter Campestre for the care and management of the animals. We also thank Prof. Dr. Patrícia Gama from the University of São Paulo for the earlier support of this project.

Authors’ contributions

TMA and VJ conceived and designed the research; TMA, FRS, and EMCO performed the experiments; TMA, FRS, WVD, and PC analyzed the data; TMA, FRS, LMR, WVD, and VJ interpreted the results of the experiments; TMA, FRS, and WVD prepared the figures; TMA and FRS drafted the manuscript; TMA, FRS, LMR, RMAM, and VJ edited and revised the manuscript. All authors read and approved the final version of the manuscript.

Compliance with ethical standards

Grant

This study was supported by grants from the Fundação de Amparo à Pesquisa do Estado de São Paulo (São Paulo State Research Foundation—grant number 2011/00036-0).

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. 1.
    Batista DG, Neves KR, Graciolli FG, dos Reis LM, Graciolli RG, Dominguez WV, Neves CL, Magalhães AO, Custódio MR, Moysés RM, Jorgetti V (2010) The bone histology spectrum in experimental renal failure: adverse effects of phosphate and parathyroid hormone disturbances. Calcif Tissue Int 87(1):60–67.  https://doi.org/10.1007/s00223-010-9367-y CrossRefPubMedGoogle Scholar
  2. 2.
    Berndt TJ, Schiavi S, Kumar R (2005) “Phosphatonins” and the regulation of phosphorus homeostasis. Am J Physiol Renal Physiol 289(6):F1170–F1182.  https://doi.org/10.1152/ajprenal.00072.2005 CrossRefPubMedGoogle Scholar
  3. 3.
    Capuano P, Radanovic T, Wagner CA, Bacic D, Kato S, Uchiyama Y, St-Arnoud R, Murer H, Biber J (2005) Intestinal and renal adaptation to a low-Pi diet of type II NaPi cotransporters in vitamin D receptor- and 1alphaOHase-deficient mice. Am J Physiol Cell Physiol 288(2):C429–C434.  https://doi.org/10.1152/ajpcell.00331.2004 CrossRefPubMedGoogle Scholar
  4. 4.
    Chavkin NW, Chia JJ, Crouthamel MH, Giachelli CM (2015) Phosphate uptake-independent signaling functions of the type III sodium-dependent phosphate transporter, PiT-1, in vascular smooth muscle cells. Exp Cell Res 333(1):39–48.  https://doi.org/10.1016/j.yexcr.2015.02.002 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Chomczynski P, Sacchi N (2006) The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on. Nat Protoc 1(2):581–585.  https://doi.org/10.1038/nprot.2006.83 CrossRefPubMedGoogle Scholar
  6. 6.
    Di Marco GS, Hausberg M, Hillebrand U, Rustemeyer P, Wittkowski W, Lang D, Pavenstädt H (2008) Increased inorganic phosphate induces human endothelial cell apoptosis in vitro. Am J Physiol Renal Physiol 294(6):F1381–F1387.  https://doi.org/10.1152/ajprenal.00003.2008 CrossRefPubMedGoogle Scholar
  7. 7.
    Duflos C, Bellaton C, Pansu D, Bronner F (1995) Calcium solubility, intestinal sojourn time and paracellular permeability codetermine passive calcium absorption in rats. J Nutr 125(9):2348–2355CrossRefPubMedGoogle Scholar
  8. 8.
    Eto N, Tomita M, Hayashi M (2006) NaPi-mediated transcellular permeation is the dominant route in intestinal inorganic phosphate absorption in rats. Drug Metab Pharmacokinet 21(3):217–221.  https://doi.org/10.2133/dmpk.21.217 CrossRefPubMedGoogle Scholar
  9. 9.
    Felsenfeld AJ, Levine BS, Rodriguez M (2015) Pathophysiology of calcium, phosphorus, and magnesium dysregulation in chronic kidney disease. Semin Dial 28(6):564–577.  https://doi.org/10.1111/sdi.12411 CrossRefPubMedGoogle Scholar
  10. 10.
    Forster I, Hernando N, Sorribas V, Werner A (2011) Phosphate transporters in renal, gastrointestinal, and other tissues. Adv Chronic Kidney Dis 18(2):63–76.  https://doi.org/10.1053/j.ackd.2011.01.006 CrossRefPubMedGoogle Scholar
  11. 11.
    Forster IC, Hernando N, Biber J, Murer H (2013) Phosphate transporters of the SLC20 and SLC34 families. Mol Asp Med 34(2-3):386–395.  https://doi.org/10.1016/j.mam.2012.07.007 CrossRefGoogle Scholar
  12. 12.
    Frei P, Gao B, Hagenbuch B, Mate A, Biber J, Murer H, Meier PJ, Stieger B (2005) Identification and localization of sodium-phosphate cotransporters in hepatocytes and cholangiocytes of rat liver. Am J Physiol Gastrointest Liver Physiol 288(4):G771–G778.  https://doi.org/10.1152/ajpgi.00272.2004 CrossRefPubMedGoogle Scholar
  13. 13.
    Giachelli CM (2003) Vascular calcification: in vitro evidence for the role of inorganic phosphate. J Am Soc Nephrol 14(90004):S300–S304.  https://doi.org/10.1097/01.ASN.0000081663.52165.66 CrossRefPubMedGoogle Scholar
  14. 14.
    Giral H, Caldas Y, Sutherland E, Wilson P, Breusegem S, Barry N, Blaine J, Jiang T, Wang XX, Levi M (2009) Regulation of rat intestinal Na-dependent phosphate transporters by dietary phosphate. Am J Physiol Renal Physiol 297(5):F1466–F1475.  https://doi.org/10.1152/ajprenal.00279.2009 CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Katai K, Miyamoto K, Kishida S, Segawa H, Nii T, Tanaka H, Tani Y, Arai H, Tatsumi S, Morita K, Taketani Y, Takeda E (1999) Regulation of intestinal Na+-dependent phosphate co-transporters by a low-phosphate diet and 1,25-dihydroxyvitamin D3. Biochem J 343(Pt 3):705–712.  https://doi.org/10.1042/bj3430705 CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Kiela PR, Ghishan FK (2009) Recent advances in the renal-skeletal-gut axis that controls phosphate homeostasis. Lab Investig 89(1):7–14.  https://doi.org/10.1038/labinvest.2008.114 CrossRefPubMedGoogle Scholar
  17. 17.
    Lau WL, Linnes M, Chu EY, Foster BL, Bartley BA, Somerman MJ, Giachelli CM (2013) High phosphate feeding promotes mineral and bone abnormalities in mice with chronic kidney disease. Nephrol Dial Transplant 28(1):62–69.  https://doi.org/10.1093/ndt/gfs333 CrossRefPubMedGoogle Scholar
  18. 18.
    Lee GJ, Marks J (2015) Intestinal phosphate transport: a therapeutic target in chronic kidney disease and beyond? Pediatr Nephrol 30(3):363–371.  https://doi.org/10.1007/s00467-014-2759-x CrossRefPubMedGoogle Scholar
  19. 19.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25(4):402–408.  https://doi.org/10.1006/meth.2001.1262 CrossRefPubMedGoogle Scholar
  20. 20.
    Magne D, Bluteau G, Faucheux C, Palmer G, Vignes-Colombeix C, Pilet P, Rouillon T, Caverzasio J, Weiss P, Daculsi G, Guicheux J (2003) Phosphate is a specific signal for ATDC5 chondrocyte maturation and apoptosis-associated mineralization: possible implication of apoptosis in the regulation of endochondral ossification. J Bone Miner Res 18(8):1430–1442.  https://doi.org/10.1359/jbmr.2003.18.8.1430 CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Mancini G, Carbonara AO, Heremans JF (1965) Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry 2(3):235–254.  https://doi.org/10.1016/0019-2791(65)90004-2 CrossRefPubMedGoogle Scholar
  22. 22.
    Mansfield K, Rajpurohit R, Shapiro IM (1999) Extracellular phosphate ions cause apoptosis of terminally differentiated epiphyseal chondrocytes. J Cell Physiol 179(3):276–286. https://doi.org/10.1002/(SICI)1097-4652(199906)179:3<276::AID-JCP5>3.0.CO;2-#Google Scholar
  23. 23.
    Marks J, Srai SK, Biber J, Murer H, Unwin RJ, Debnam ES (2006) Intestinal phosphate absorption and the effect of vitamin D: a comparison of rats with mice. Exp Physiol 91(3):531–537.  https://doi.org/10.1113/expphysiol.2005.032516 CrossRefPubMedGoogle Scholar
  24. 24.
    Marks J, Churchill LJ, Srai SK, Biber J, Murer H, Jaeger P, Debnam ES, Unwin RJ, Group ETaCB (2007) Intestinal phosphate absorption in a model of chronic renal failure. Kidney Int 72:166–173.  https://doi.org/10.1038/sj.ki.5002292 CrossRefPubMedGoogle Scholar
  25. 25.
    Marks J, Churchill LJ, Debnam ES, Unwin RJ (2008) Matrix extracellular phosphoglycoprotein inhibits phosphate transport. J Am Soc Nephrol 19(12):2313–2320.  https://doi.org/10.1681/ASN.2008030315 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Marks J, Debnam ES, Unwin RJ (2010) Phosphate homeostasis and the renal-gastrointestinal axis. Am J Physiol Renal Physiol 299(2):F285–F296.  https://doi.org/10.1152/ajprenal.00508.2009 CrossRefPubMedGoogle Scholar
  27. 27.
    Marks J, Lee GJ, Nadaraja SP, Debnam ES, Unwin RJ (2015) Experimental and regional variations in Na+-dependent and Na+-independent phosphate transport along the rat small intestine and colon. Physiol Rep 3(1):e12281.  https://doi.org/10.14814/phy2.12281 CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Meleti Z, Shapiro IM, Adams CS (2000) Inorganic phosphate induces apoptosis of osteoblast-like cells in culture. Bone 27(3):359–366.  https://doi.org/10.1016/S8756-3282(00)00346-X CrossRefPubMedGoogle Scholar
  29. 29.
    Murer H, Biber J (1996) Molecular mechanisms of renal apical Na/phosphate cotransport. Annu Rev Physiol 58(1):607–618.  https://doi.org/10.1146/annurev.ph.58.030196.003135 CrossRefPubMedGoogle Scholar
  30. 30.
    Park JW, Yook JM, Ryu HM, Choi SY, Morishita M, Do JY, Park SH, Kim CD, Choi JY, Chung HY, Kim YL (2011) Phosphate-induced apoptosis in human peritoneal mesothelial cells in vitro. Am J Nephrol 34(1):77–86.  https://doi.org/10.1159/000329081 CrossRefPubMedGoogle Scholar
  31. 31.
    Picard N, Capuano P, Stange G, Mihailova M, Kaissling B, Murer H, Biber J, Wagner CA (2010) Acute parathyroid hormone differentially regulates renal brush border membrane phosphate cotransporters. Pflugers Arch 460(3):677–687.  https://doi.org/10.1007/s00424-010-0841-1 CrossRefPubMedGoogle Scholar
  32. 32.
    Ritter CS, Finch JL, Slatopolsky EA, Brown AJ (2001) Parathyroid hyperplasia in uremic rats precedes down-regulation of the calcium receptor. Kidney Int 60(5):1737–1744.  https://doi.org/10.1046/j.1523-1755.2001.00027.x CrossRefPubMedGoogle Scholar
  33. 33.
    Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T (2006) The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol 7(1):3.  https://doi.org/10.1186/1471-2199-7-3 CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Sommer S, Berndt T, Craig T, Kumar R (2007) The phosphatonins and the regulation of phosphate transport and vitamin D metabolism. J Steroid Biochem Mol Biol 103(3-5):497–503.  https://doi.org/10.1016/j.jsbmb.2006.11.010 CrossRefPubMedGoogle Scholar
  35. 35.
    Virkki LV, Biber J, Murer H, Forster IC (2007) Phosphate transporters: a tale of two solute carrier families. Am J Physiol Renal Physiol 293(3):F643–F654.  https://doi.org/10.1152/ajprenal.00228.2007 CrossRefPubMedGoogle Scholar
  36. 36.
    Zegre Cannon C, Kissling GE, Goulding DR, King-Herbert AP, Blankenship-Paris T (2011) Analgesic effects of tramadol, carprofen or multimodal analgesia in rats undergoing ventral laparotomy. Lab Anim (NY) 40(3):85–93.  https://doi.org/10.1038/laban0311-85 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Tatiana Martins Aniteli
    • 1
  • Flávia Ramos de Siqueira
    • 1
  • Luciene Machado dos Reis
    • 1
  • Wagner Vasques Dominguez
    • 1
  • Elizabeth Maria Costa de Oliveira
    • 1
  • Patrícia Castelucci
    • 3
  • Rosa Maria Affonso Moysés
    • 1
    • 2
  • Vanda Jorgetti
    • 1
    • 4
    Email author
  1. 1.Medical School, Division of NephrologyUniversidade de São PauloSão PauloBrazil
  2. 2.Universidade Nove de Julho – UNINOVESão PauloBrazil
  3. 3.Department of Anatomy, Institute of Biomedical SciencesUniversidade de São PauloSão PauloBrazil
  4. 4.Faculdade de Medicina, Serviço de NefrologiaUniversidade de São PauloSão PauloBrazil

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