Effects Of Fucoidan On The Accumulation Of Extracellular Matrix In Human Mesangial Cells (HMC) Which Induced By Growth Factor Betal

(Objective] To investigate the effective of Fucoidan(FPS) on the secretion of Fibronectin(FN)、matrix metalloproteinases-2(MMP-2) and tissue inhibitor of metalloproteinases-2(TIMP-2) and phosphorylation of ERK1/2in human mesangial cells (HMC) which induced by growth factor β1,so we can elplore how FPS prevents the accumulation of Extracellular Matrix in HMC.[Methods] HMC were cultured in vitro with TGF-β1, and treated with different concentration of FPS. The cells were divided into6groups:model group(TGF-β1:4ng/ml), high-FPS group (TGF-(31:4ng/ml+FPS:100ug/ml), medium-FPS group (TGF-β1:4ng/ml+FPS:50ug/ml),low-FPS group (TGF-β1:4ng/ml+FPS:25ug/ml), FPS control group (FPS:100ug/ml) and blank control group. The level of MMP-2and TIMP-2protein expression was measured by ELISA, The level of MMP-2、TIMP-2and FN mRNA expression was measured by Resltime-PCR. The level of FN、P-ERK1/2、ERK protein expression was measured by Westernblot.[Results](1) Through immunofluorescence method we abserved the cellular markers of the cells such as actin, myosin, vimentin, desmin were expressed,and the expressions of factor Ⅷ and cytokeratin were negative, which can prove that the cells were HMC.(2)With the stimulation of TGF-β1, the expression of FN protein and mRNA was increased significantly compared with blank control group (P<0.01), which then were decreased by the treatment of FPS (P<0.01P<0.05). Compared with blank control group, the expression of FN protein and mRNA in FPS-control group was not statistically significant (P>0.05).(3) With the stimulation of TGF-β1, the expression of MMP-2protein and mRNA was decreased significantly compared with blank control group (P<0.01), which then were increased by the treatment of FPS (P<0.01), and showed dose dependent manner; the expression of TIMP-2protein and mRNA was increased significantly compared with blank control group (P<0.01), which then were decreased by the treatment of FPS (P<0.01), and showed dose dependent manner too. Compared with blank control group, the expression of MMP-2and TIMP-2protein and mRNA in FPS-control group was not statistically significant (P>0.05).(4)With the stimulation of TGF-β1, the expression of P-ERK1/2protein was increased significantly compared with blank control group (P<0.01), which then were decreased by the treatment of FPS (P<0.01P<0.05), and showed dose dependent manner. Compared with blank control group, the expression of P-ERK1/2protein in FPS-control group was not statistically significant (P>0.05).[Conclusion](1)High purity HMC was cultivated and that made a basis for research of cytokines express in HMC which cultured with TGF-β1and FPS.(2)The expression of FN in HMC may be induced by TGF-β1and FPS can inhibit it.Thus, the accumulation of Extracelluar matrix (ECM) and glomerulosclerosis can be prevented.(3)The disbananlance of MMP-2and TIMP-2in HMC may be induced by TGF-β1and FPS can block the alteration. Thus, the accumulation of Extracelluar Matrix (ECM) and glomerulosclerosis can be prevented.(4)The phosphorylation of ERK1/2in HMC may be induced by TGF-β1and FPS can inhibit it. Thus, the accumulation of Extracelluar matrix (ECM) and glomerulosclerosis can be prevented.