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Objective:To investigate the effects of Herbal Compound 861(Cpd 861) on collagen synthesis and degradation in rat mesangial cells exposed to high glucose.Methods:The third to fifth passage of rat mesangial cells were exposed to high glucose and Cpd 861 at a concentration of 0.25-4.00 g/L for 24,48and 72 h,respectively.Benazepril(10~(-7)-10~(-3) mmol/L) was selected as positive control.The methyl thiazolyl tetrazolium colorimetric assay was used to evaluate the effect of Cpd 861 on cell proliferation.After incubation with Cpd 861 at a concentration of 2.00 g/L for 48 h,the protein secretions of collagen type Ⅳ,matrix metallopeptidase 9(MMP-9),tissue inhibitor of metalloproteinase-1(TIMP-1),transforming growth factor beta1(TGF-β_1),and hepatocyte growth factor(HGF) were detected by enzyme-linked immunosorbent assay method.And rat mesangial cells were harvested to determine MMP-9,TIMP-1,TGF-β_1 and HGF mRNA expression by reverse transcription polymerase chain reaction.Results:Cpd 861 inhibited cell proliferation induced by high glucose in a dose- and time-dependent manner.Compared with high glucose,collagen typeⅣ production was decreased significantly by Cpd 861(P<0.01).Cpd 861 increased the protein secretions and mRNA expressions of MMP-9 and HGF,whereas the protein secretions and mRNA expressions of TIMP-1and TGF-β_1 were reduced markedly(P<0.05).The ratio of MMP-9 to TIMP-1 was enhanced by Cpd 861significantly.There was no significant difference in all above-mentioned effects between Cpd 861(2.00 g/L)and benazepril(10~(-5) mmol/L).Conclusion:The anti-glomerulosclerosis mechanisms of Cpd 861 were partly attributed to its effects of inhibiting mesangial cell proliferation,decreasing collagen synthesis and enhancing collagen degradation.
Objective: To investigate the effects of Herbal Compound 861 (Cpd 861) on collagen synthesis and degradation in rat mesangial cells exposed to high glucose. Methods: The third to fifth passage of rat mesangial cells were exposed to high glucose and Cpd 861 at a concentration of 0.25-4.00 g / L for 24,48 and 72 h respectively.Benazepril (10 -7 -10 -3 mmol / L) was selected as positive control.The methyl thiazolyl tetrazolium colorimetric assay was used to evaluate the effect of Cpd 861 on cell proliferation. After incubation with Cpd 861 at a concentration of 2.00 g / L for 48 h, the protein secretions of collagen type IV, matrix metallopeptidase 9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), transforming growth factor beta 1 (TGF-beta 1), and hepatocyte growth factor (HGF) were detected by enzyme-linked immunosorbent assay method. -β 1 and HGF mRNA expression by reverse transcription polymerase chain reaction. Results: Cpd 861 inhibited cell proliferation induced by high glucose in a dose- and time-dependent manner. Compared with high glucose, collagen type IV production was significantly significantly decreased by Cpd 861 (P <0.01) .Cpd 861 increased the protein secretions and mRNA expressions of MMP-9 and the HGF, whereas the protein secretions and mRNA expressions of TIMP-1 and TGF-β_1 were reduced markedly (P <0.05). The ratio of MMP-9 to TIMP-1 was enhanced by Cpd 861 slightlyificly.There was no significant difference in all above -cative effects between Cpd 861 (2.00 g / L) and benazepril (10-5 mmol / L) .Conlusion: The anti-glomerulosclerosis mechanisms of Cpd 861 were partly attributed to its effects of inhibiting mesangial cell proliferation, reducing collagen synthesis and enhancing collagen degradation.