| Objectives:To investigate the inhibition effect of the serum in radix astragali, hirudo and the compoundings with effective constituent in vitro rat mesangial cells (MC) proliferation, study the effect of yiqi huayu medicine on the PDGF-BB expression correlation in rat glomerular mesangial cells(GMCs) under LPS, compare the different effect strength with serum in radix astragali, hirudo and the compoundings with effective constituent, explore the function and possible inhibition mechanism of yiqi huayu medicines on proliferation of mesangial cells.Methods:Rat glomerular mesangial cells were cultured with cell culture technology in vitro, and the radix astragali, hirudo and the compoundings contained serum of model rats were prepared by adopting the method of serum pharmacology. Then GMCs were cultured with radix astragali high dose, hirudo high dose, high radix astragali and high hirudo dose, low radix astragali and high hirudo dose, LPS model group, astragalus polyose group, astragaloside IV group, calycosin group, hirudin group, the compoundings group and at the same time set up normal control group. Add the drug-containing serum and the compoundings with effective constituent induced in vitro culture GMCs under LPS, detect the proliferation effect of glomerular mesangial cell CCK-8 method, and respectively detected the mRNA and protein expression of PDGF-BB of yiqi huayu Medicine in rat glomerular mesangial cells under LPS.Results:(1) The normal control group can express PDGF-BB; (2) Compared with normal control group, the expressions of PDGF-BB protein and mRNA can obviously increased (P< 0.05) under the stimulation of LPS; (3) Compared with LPS group, different serum and the compoundings with effective constituent of Yiqi Huayu medicine could significantly inhibited the expression of PDGF-BB mRNA and protein(P<0.05); (4) Compared with radix astragali high dose group, the PDGF-BB mRNA expression of astragalus polyose group had significantly increased (P<0.01); Compared with hirudo high dose group, high radix astragali and high hirudo dose group and hirudin group had strong inhibition (P<0.05), the inhibitory effect of low astragalus with hirudo high dose is not obvious (P>0.05), (5) The serum and the compoundings with effective constituent of yiqi huayu medicine can different effect the proliferation of GMCs, the inhibitory effect of each hirudo contained group is more significant(P<0.05), and increased with the increase of astragalus, hirudo dose (P<0.05), and it showed a certain dose-dependent (P<0.05).Conclusions:(1) Lipopolysaccharide(LPS) could stimulated the proliferation of GMCs, resulted in the increased expression of PDGF-BB mRNA and protein in mesangial cells, prompted that PDGF-BB participated in the pathogenesis of renal damage process in the mesangial proliferative glomerulonephritis (MsPGN). (2)Different serum and the compoundings with effective constituent of yiqi huayu medicine contained can reduce the expression of PDGF-BB mRNA and protein, Compared with astragalus contained serum group and its effective components, that PDGF-BB mRNA expression of astragalus polysose had significantly increased than radix astragali high dose, illustrated the effenct of astragalus effective components are no better than drug-contained serum; it conformed that hirudin group had stronger inhibitory effect by the comparison of hirudo contained medicine serum and hirudin group, however, whether contained medicine serum or active ingredients, the compoundings with effective constituent mainly composed of radix astragali is better than hirudo, the study also showed that research mainly on the serum of yiqi huayu medicine while not at the active ingredients of single application, which was beneficial to decline the progress of glomerular sclerosis of MsPGN. (3)To explore the possible inhibit mechanism of yiqi huayu medicines on proliferation of mesangial cells from molecular biology level, provide science experiment basis for clinical application of yiqi huayu chinese medicine. |
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