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Vol. 101, No. 4, 2005
Issue release date: December 2005
Nephron Exp Nephrol 2005;101:e119–e126

Tacrolimus and Cyclosporinein vitro and in vivo Induce Osteopontin mRNA and Protein Expression in Renal Tissues

Khanna A.
Department of Medicine (Nephrology), Medical College of Wisconsin, Milwaukee, Wisc., USA

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The mechanism of immunosuppression-linked nephrotoxicity in organ transplantation remains to be solved. Expression of osteopontin (OPN), a multifunctional secreted glycoprotein, has been associated with various forms of renal injuries. In this study, using in vitro and in vivo models, we examined the effects of cyclosporine (CsA) and tacrolimus (TAC) on OPN mRNA and protein expression. We also examined if CsA- and TAC-induced OPN expression is dependent on transforming growth factor (TGF)-β expression. For in vivo experiments mice and rats were injected with CsA (25 mg/kg) and TAC (0.2 mg/kg). For in vitro experiments, human proximal tubular epithelial (PTE) cells were treated with CsA and TAC for 4 h. To study the in vivo effect of TGF-β on OPN mRNA, mice were injected with recombinant TGF-β protein (3 mg/kg). The expression of OPN was also studied in CsA-treated PTE cells with and without anti-TGF-β antibody. At the end of in vitro and in vivo treatments, RNA was isolated from kidney tissue and renal cells reverse transcribed to cDNA and amplified for OPN mRNA. Using immunochemistry and Western blot analysis OPN protein expression was also studied in vivo and in vitro, respectively. Both in vitro and in vivo treatment with CsA and TAC resulted in significantly increased OPN mRNA and protein expression. TGF-β treatment in vivo also resulted in a significantly increased OPN mRNA expression and anti-TGF-β antibody but not the control antibody in vivo in CsA-treated mice, and in vitro in CsA-treated PTE cells inhibited OPN mRNA expression. OPN may contribute to the CsA- and TAC-induced nephrotoxicity in organ transplant recipients and the increased OPN expression might be mediated by TGF-β.

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  1. O’Brien ER, Garvin MR, Stewart DK, Hinohara T, Simpson JB, Schwartz SM, et al: Osteopontin is synthesized by macrophage, smooth muscle, and endothelial cells in primary and restenotic human coronary atherosclerotic plaques. Arterioscler Thromb 1994;14:1648–1656.
  2. O’Regan AW, Chupp GL, Lowry JA, Goetschkes M, Mulligan N, Berman JS: Osteopontin is associated with T cells in sarcoid granulomas and has T cell adhesive and cytokine-like properties in vitro. J Immunol 1999;162:1024–1031.
  3. Malyankar UM, Almeida M, Johnson RJ, Pichler RH, Giachelli CM: Osteopontin regulation in cultured rat renal epithelial cells. Kidney Int 1997;51:1766–1773.
  4. Hu DD, Lin EC, Kovach NL, Hoyer JR, Smith JW: A biochemical characterization of the binding of osteopontin to integrins alpha v beta 1 and alpha v beta 5. J Biol Chem 1995;270:26232–26238.
  5. Singh K, Deonarine D, Shanmugam V, Senger DR, Mukherjee AB, Chang PL, Prince CW, Mukherjee BB: Calcium-binding properties of osteopontin derived from non-osteogenic sources. J Biochem 1993;114:702–707.
  6. Wuthrich RP: The complex role of osteopontin in renal disease. Nephrol Dial Transplant1999;13:2448–2450.
  7. Panzer U, Thaiss F, Zahner G, Barth P, Reszka M, Reinking RR, Wolf G, Helmchen U, Stahl RA: Monocyte chemoattractant protein-1 and osteopontin differentially regulate monocytes recruitment in experimental glomerulonephritis. Kidney Int2001;59:1762–1769.
  8. Yu XQ, Nikolic-Paterson DJ, Mu W, Giachelli CM, Atkins RC, Johnson RJ, Lan HY: A functional role for osteopontin in experimental crescentic glomerulonephritis in the rat. Proc Assoc Am Physicians 1998;110:50–64.
  9. Mazzali M, Kipari T, Ophascharoensuk V, Wesson JA, Johnson R, Hughes J: Osteopontin – a molecule for all seasons. QJM 2002;95:3–13.
  10. Xie Y, Sakatsume M, Nishi S, Narita I, Arakawa M, Gejyo F: Expression, roles, receptors, and regulation of osteopontin in the kidney. Kidney Int 2001;60:1645–1657.
  11. Suthanthiran M: Renal transplantation: Reaping the rewards of biomedical research. Curr Opin Nephrol Hypertens 2000;9:597–598.
  12. Khanna A, Cairns V, Hosenpud JD: Tacrolimus induces increased expression of transforming growth factor-beta1 in mammalian lymphoid as well as nonlymphoid cells. Transplantation 1999;67:614–619.
  13. Khanna AK, Li B, Stenzel KH, Suthanthiran M: Regulation of new DNA synthesis in mammalian cells by cyclosporine: demonstration of a transforming growth factor-β dependent mechanism of inhibition of cell growth. Transplantation 1994;57:577–582.
  14. Khanna A, Cairns V, Becker CG, Hosenpud JD: Transforming growth factor-β (TGF-β) mimics and anti-TGF-β antibody abrogates the in-vivo effects of cyclosporine: demonstration of a direct role of TGF-β in immunosuppression and nephrotoxicity of CsA. Transplantation 1999;67:882–889.
  15. Zhou H, Hammonds RG Jr, Findlay DM, Martin TJ, Ng KW: Differential effects of transforming growth factor-beta 1 and bone morphogenetic protein 4 on gene expression and differentiated function of preosteoblasts. J Cell Physiol 1993;155:112–119.
  16. Oldberg A, Franzen A, Heinegard D: Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc Natl Acad Sci USA 1986;83:8819–8823.
  17. Kiefer MC, Bauer DM, Barr PJ: The cDNA and derived amino acid sequence for human osteopontin. Nucleic Acids Res 1989;17:3306.
  18. Miyazaki Y, Setoguchi M, Yoshida S, Higuchi Y, Akizuki S, Yamamoto S: The mouse osteopontin gene. Expression in monocytic lineages and complete nucleotide sequence. J Biol Chem 1990;265:14432–14438.
  19. Xie Y, Nishi S, Iguchi S, Imai N, Sakatsume M, Saito A, Ikegame M, Iino N, Shimada H, Ueno M, Kawashima H, Arakawa M, Gejyo F: Expression of osteopontin in gentamicin-induced acute tubular necrosis and its recovery process. Kidney Int 2001;59:959–974.
  20. Hudkins KL, Le QC, Segerer S, Johnson RJ, Davis CL, Giachelli CM, Alpers CE: Osteopontin expression in human cyclosporine toxicity. Kidney Int 2001;60:635–640.
  21. Khanna A, Plummer M, Bromberek C, Bresnahan B, Hariharan S: Expression of TGF-β and fibrogenic genes in transplant recipients with Tacrolimus and cyclosporine nephrotoxicity. Kidney Int 2002;62:2257–2263.
  22. Mezzano SA, Barria M, Droguett MA, Burgos ME, Ardiles LG, Flores C, Egido J: Tubular NF-kappaB and AP-1 activation in human proteinuric renal disease. Kidney Int 2001;60:1366–1377.
  23. Tamada S, Nakatani T, Asai T, Tashiro K, Komiya T, Sumi T, Okamura M, Kim S, Iwao H, Kishimoto T, Yamanaka S, Miura K: Inhibition of nuclear factor-kappaB activation by pyrrolidine dithiocarbamate prevents chronic FK506 nephropathy. Kidney Int 2003;63:306–314.
  24. Asai T, Nakatani T, Yamanaka S, Tamada S, Kishimoto T, Tashiro K, Nakao T, Okamura M, Kim S, Iwao H, Miura K: Magnesium supplementation prevents experimental chronic cyclosporine a nephrotoxicity via renin-angiotensin system independent mechanism. Transplantation 2002;74:784–791.
  25. Young BA, Burdmann EA, Johnson RJ, Alpers CE, Giachelli CM, Eng E, Andoh T, Bennett WM, Couser WG: Cellular proliferation and macrophage influx precede interstitial fibrosis in cyclosporine nephrotoxicity. Kidney Int 1995;48:439–448.
  26. Lee SK, Park JY, Yu ES, Yang WS, Kim SB, Park SK, Park JS: Individual or combined effects of enalapril and verapamil on chronic cyclosporine nephrotoxicity in rats. J Korean Med Sci 1999;14:653–658.
  27. Lewington AJ, Padanilam BJ, Martin DR, Hammerman MR: Expression of CD44 in kidney after acute ischemic injury in rats. Am J Physiol Regul Integr Comp Physiol 2000;278:R247–R254.
  28. Johnson RJ, Gordon KL, Giachelli C, Kurth T, Skelton MM, Cowley AW Jr: Tubulointerstitial injury and loss of nitric oxide synthases parallel the development of hypertension in the Dahl-SS rat. J Hypertens 2000;18:1497–1505.
  29. Diamond JR, Kreisberg R, Evans R, Nguyen TA, Ricardo SD: Regulation of proximal tubular osteopontin in experimental hydronephrosis in the rat. Kidney Int 1998;54:1501–1509.
  30. Abbate M, Zoja C, Corna D, Capitanio M, Bertani T, Remuzzi G: In progressive nephropathies, overload of tubular cells with filtered proteins translates glomerular permeability dysfunction into cellular signals of interstitial inflammation. J Am Soc Nephrol1998;9:1213–1224.
  31. Pichler R, Giachelli CM, Lombardi D, Pippin J, Gordon K, Alpers CE, Schwartz SM, Johnson RJ: Tubulointerstitial disease in glomerulonephritis. Potential role of osteopontin (uropontin). Am J Pathol 1994;144:915–926.
  32. Mazzali M, Hughes J, Dantas M, Liaw L, Steitz S, Alpers CE, Pichler RH, Lan HY, Giachelli CM, Shankland SJ, Couser WG, Johnson RJ: Effects of cyclosporine in osteopontin null mice. Kidney Int 2002;62:78–85.
  33. Lim BJ, Kim PK, Hong SW, Jeong HJ: Osteopontin expression and microvascular injury in cyclosporine nephrotoxicity. Pediatr Nephrol 2004;19:288–294.

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