Effect Of The Pyridoxamine On The Cell Proliferation And The Expressions Of TGF-β1,MMP-2,TIMP-2 And Fn In Rat Glomerular Mesangial Cells Under High-glucose Condition

Object: To observe the effect of different concentrations of the pyridoxamine on the cell proliferation, the dynamic changes of transforming growth factor-β1, matrix metalloproteinase-2, tissue inhibitor of metalloproteinase-2 and fibronectin secreted by rat glomerular mesangial cells cultured in high glucose. To investigate the protective effect of pyridoxamine in patient with diabetic nephropathy, and further to explore the possible mechanisms.Methods: Rat glomerular mesangial cells in vitro were cultured in different concentrations glucose. The mesangial cells treated by high glucose were exposed to the pyridoxamine in different doses and time. The cell proliferation of mesangial cells was measured by MTT colorimetric method. The expressions of transforming growth factor-β1, matrix metalloproteinase-2, tissue inhibitor of metalloproteinase-2 and fibronectin mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction. The levels of transforming growth factor-β1 and fibronectin protein in the supernatant were assessed by enzyme linked immunosorbent assay. The contents of malondialdehyde in supernatants were evaluated by spectrophotometer.Results: The high level of glucose concentration could promote the cell proliferation of mesangial cells in the range of 24 to 72 hours. The high level of glucose could stimulate the expressions of transforming growth factor-β1, tissue inhibitor of metalloproteinase-2 and fibronectin mRNA, the contents of malondialdehyde and the levels of transforming growth factor-β1 and fibronectin protein in the supernatant, and also could decrease the expressions of the matrix metalloproteinase-2 mRNA. The pyridoxamine had a significant inhibition of the cell proliferation and the expressions of transforming growth factor-β1, tissue inhibitor of metalloproteinase-2 and fibronectin mRNA of mesangial cells cultured in high glucose, and could also decrease the contents of malondialdehyde and the levels of transforming growth factor-β1 and fibronectin protein in the supernatant. The pyridoxamine could promote the matrix metalloproteinase-2 mRNA. The influences were associated with the doses of the pyridoxamine.Conclusions: High glucose stimulates the cell proliferation of mesangial cells in some concentration and time range. High glucose contributes to an increasing of malondialdehyde and the transforming growth factor-β1 produced by mesangial cells and imbalance between the synthesis and degradation of fibronectin and matrix metalloproteinase-2 / tissue inhibitor of metalloproteinase-2. The pyridoxamine attenuates these effects of high glucose on mesangial cells in a dose-dependent manner.