Tacrolimus and cyclosporinein vitro and in vivo induce osteopontin mRNA and protein expression in renal tissues.
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)-beta 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-beta on OPN mRNA, mice were injected with recombinant TGF-beta protein (3 mg/kg). The expression of OPN was also studied in CsA-treated PTE cells with and without anti-TGF-beta 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-beta treatment in vivo also resulted in a significantly increased OPN mRNA expression and anti-TGF-beta 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-beta.
Khanna
Ashwani
A
Department of Medicine (Nephrology), Medical College of Wisconsin, Milwaukee, WI 53226, USA. akkhanna@mcw.edu
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D016572:Cyclosporine / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000737:chemistry* / Q000166:cytology / Q000187:drug effects; D051379:Mice; D008815:Mice, Inbred Strains; D053495:Osteopontin; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics*; D016559:Tacrolimus / Q000494:pharmacology*; D016212:Transforming Growth Factor beta / Q000276:immunology / Q000494:pharmacology
D000906:Antibodies; D012333:RNA, Messenger; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D016212:Transforming Growth Factor beta; D053495:Osteopontin; D016572:Cyclosporine; D016559:Tacrolimus