Effects of mycophenolate mofetil on kidney function and phosphorylation status of renal proteins in Alport COL4A3-deficient mice

Abstract

BackgroundWe investigated the effects of mycophenolate mofetil (MMF) on kidney function and on protein phosphorylation in a mouse model for the human Alport syndrome.MethodsCOL4A3-deficient (COL4A3−/−) mice were randomly allocated to receive a placebo (PLC COL4A3−/−) or MMF treatment (MMF COL4A3−/−). Wild type mice (WT) were used as controls. Changes in serum creatinine, total protein and blood urea nitrogen (BUN), concentrations of mycophenolic acid (MPA) and its glucuronide metabolite (MPAG), serum protein electrophoresis, urine dipstick chemistry and sediment were measured. Changes in the phosphorylation status of renal proteins and histology were analyzed.ResultsMMF influenced kidney function and protein phosphorylation. Serum creatinine and BUN were lower in MMF treated compared to PLC treated COL4A3−/− mice. Serum albumin and alpha-1 globulins were significantly decreased while serum creatinine, alpha-2 globulins, urine dipstick protein, leukocyte esterase, hemoglobin and red blood cells were all increased in both COL4A3−/− groups compared to WT. Differential 2DE-gel analysis identified six phosphorylated kidney protein spots that were significantly altered by MMF.ConclusionsThese data suggest that the MMF treatment in this murine model moderately improved kidney function and reversed the phosphorylation status of six renal phosphoprotein spots to that seen in WT mice.Electronic supplementary materialThe online version of this article (doi:10.1186/s12953-014-0056-z) contains supplementary material, which is available to authorized users.

Mycophenolic acid reverses TGF beta‐induced cell motility, collagen matrix contraction and cell morphology in vitro

“…We concluded that MPA showed antifibrotic effects in vitro. Furthermore, these in vitro results correlate well with a previously published in vivo study, 15 where MMF treatment of COL4A3-defficient mice was associated with moderately improved kidney function and the reversed phosphorylation status of six renal phosphoprotein spots compared to that seen in healthy wild-type mice. The involved proteins were associated with a number of important cell properties including: cell projection, motility, migration, invasion, polarity, division, transformation and cell growth (T cells, B cells and fibroblasts).…”

“…14,9 Our recent in vivo study on the effects of MPA in COL4A3 deficient mice suggested moderately improved kidney function, presumably through inhibition of tubulointerstitial fibrosis and through regulation of phosphoproteins involved in cell projection, motility, migration, invasion, polarity, division, transformation and/or cell growth. 15 Our hypothesis is that MPA can inhibit renal fibrosis by disturbing TGF transduction and changing the cell phenotype. Therefore, we designed the current in vitro study using a human HK-2 cell line 16 stimulated by EGF and/or TGF.…”

“…TGF‐independent signal pathways have also been suggested to play a role in tubulointerstitial fibrosis . Our recent in vivo study on the effects of MPA in COL4A3 deficient mice suggested moderately improved kidney function, presumably through inhibition of tubulointerstitial fibrosis and through regulation of phosphoproteins involved in cell projection, motility, migration, invasion, polarity, division, transformation and/or cell growth …”