Inhibitory Effect Of Heparin On Rat Glomerulus Mesangial Cell Proliferation And Activation Of Nuclear Factor-κB

PrefaceGlomerular mesangial cells (GMCs) proliferation play significant pathogen-esis roles in the development of glomerular injury. Some findings demonstrared that nuclear factor - kappa B ( NF - KB ) appear to be important in cell proliferation. Recently, there are increasing evidences that suggest that heparin can inhibit GMCs proliferation. We hypothesized that heparin may decrease GMCs proliferation through the inhibition of NF - kB activation. To address this issue, we studied the effects of heparin on GMCs proliferation and activation of NF -KB.Materials and Methods1. The rat GMCs were bought from Shanghai Changzheng Hospital and cultured using the methods described by reference.2. GMCs were cultured in 96 well plates. Three group were established: (1) GMCs group; (2) lipopolysaccharide (LPS) group (GMCs + UPS) ; (3) heparin group (GMCs + LPS + heparin) , in which there are different concentrations of heparin (3 g/ml,30 g/ml ,300 g/ml). GMCs proliferation was detected by MTT at 24hs after cultured.3. GMCs were cultured in 25 cm2 culture flasks. Three group were established: (1) GMCs group; (2) lipopolysaccharide (LPS) group (GMCs + LPS) ; (3) heparin group (GMCs + LPS + heparin) , the terminal concentrations of heparin is 300 g/ml. After GMCs were cultured 24 hours, a nuclear protein extract was obtained. Immunoblotting was applied to examine the expression of p65 in nuclear.4. Statistical analysisResults were expressed as mean SD ( x s ). Comparisions among 3 groups were analyzed by one - way analysis of variance, followed by LSD multiple comparision test to evaluate statistical difference between 2 groups (SPSS12. 0 statistical software) , p <0.05 is considered as significant.Results1. Effct of heparin on GMCs proliferationIn this study, heparin was found to inhibit GMCs proliferation in a dose -dependent relationship with the concentration.2. Effct of heparin on NF - KB acivationThe result of western blot showed that the NF - KB expression was obviously higher in the LPS group than in the GMCs group, while it was obviously lower in the haparin group than that in the LPS group.Conclusion1. Heparin can inhibit GMCs proliferation.2. Heparin can inhibit the activation of NF - KB.