Effect And Mechanisms Of Flavonoids From The Leaves Of Carya Cathayensis Sarg On Proliferation And Adipogenic Differentiation Of C3H10T1/2 Cells

Objective:In This study we aims to investigate the effcet of total flavonoid (TFs) which was isolated from the leaves of Carya cathayensis Sarg., and five flavonoids, i.e. cardamonin, pinostrobin chalcone (PC), wogonin, chrysin and pinocembrin on proliferation and adipogenic differentiation of C3H10T1/2 cells, and screening the components to inhibit adipogenesis.Furthermore,to clear the preliminary mechanisms, in order to provides an experimental basis for preventing and treating diseases caused by obesity.Methods:(1) Establishment the model of C3H10T1/2 cells differentiated into adipocytes,and its identification.(2) The effect of TFs and five flavonoids on the growth of C3H10T1/2 cells was detected by MTS assay.(3) The effect of TFs and five flavonoids on proliferation and adipogenic differentiation of C3H10T1/2 cells was detected by oil red O staining.(4) The triglyceride content of TFs and five flavonoids on proliferation and adipogenic differentiation of C3H10T1/2 cells was detected by GPO-PAPenzymatic.(5) Expression of PPARγ,C/EBPα,FABP4 mRNA after treatment of C3H10T1/2 cells with TFs, cardamonin, pinostrobin chalcone (PC), chrysin and pinocembrin, were detected by fluorescent quantitative PCR.(6) The protein expression of PPARγ,C/EBPα,FABP4 that C3H10T1/2 cells were incubated with TFs, cardamonin, pinostrobin chalcone (PC), chrysin and pinocembrin, were detected by Western-blot.Results:(1) Classics appraisal, using C3H10T1/2 cells can successfully established model of cell adipogenic differentiation.(2) TFs, chrysin, wogonin, cardamonin, pinocembrin and PC in 30μg/mL,60μmol/L,60μmol/L,50μmol/L,60μmol/L,60μmol/L concentrations, respectively, inhibited C3H10T1/2 cells growth(P<0.05orP<0.01)in a dose-dependent manner.(3)TFs,chrysin,wogonin,cardamonin,pinocembrin and PC in 10μg/mL,30μmol/L,5μmol/L,30μmol/L,30μmol/L concentrations, respectively,inhibit-ed C3H10T1/2 adipocyte differentiation(P<0.05 or P<0.01) in a dose-dependent manner; wogonin can’t inhibited C3H10T1/2 adipocyte differentiation. (4) TFs, chrysin, wogonin,cardamonin,pinocembrin and PC in 10μg/mL,30μmol/L,5μmol/L, 30μmol/L,30μmol/L concentrations, respectively, inhibited the accumulation of triglyceride in the cytoplasm of adipocytes(P<0.05orP<0.01) in a dose-dependent manner. wogonin can’t inhibited inhibited the accumulation of triglyceride in the cytoplasm of adipocytes. (5) TFs,chrysin,wogonin,cardamonin,pinocembrin and PC in 5μg/mL,10μmol/L,10μmol/L,10μmol/L,50μmol/L concentrations, respectively, down-regulated the mRNA levels of PPARy and C/EBPa in C3H10T1/2 cells(P<0.05 orP<0.01) in a dose-dependent manner; TFs, chrysin, wogonin, cardamonin, pinoce-mbrin and PC in10μg/mL,5μmol/L,10μmol/L,5μmol/L,50μmol/L concentrations, respectively,down-regulated the mRNA levels of FABP4 in C3H10T1/2 cells(P<0.05 orP<0.01)in a dose-dependent manner. (6) TFs,chrysin,wogonin,cardamonin,pinocem-brin and PC in 20μg/mL,10μmol/L,5μmol/L,50μmol/L,50μmol/L concentrations, respectively, down-regulated the levels of PPARy and C/EBPa protein in C3H10T1/2 cells(P<0.05orP<0.01) in a dose-dependent manner; TFs,chrysin,wogonin,cardamom-mnin, pinocembrin and PC in10μg/mL,10μmol/L,5μmol/L,30μmol/L,30μmol/L concentrations, respectively, down-regulated the levels of FABP4 protein in C3H10T1/2 cells (P<0.05orP<0.01) in a dose-dependent manner.Conclusion:(1) TFs, chrysin, cardamonin, pinocembrin and PC can inhibited C3H10T1/2 adipocyte differentiation, wogonin can’t inhibited C3H10T1/2 adipocyte differentiation. (2) TFs,chrysin,cardamonin,pinocembrin and PC can inhibited the accumulation of triglyceride in the cytoplasm of adipocytes, wogonin can’t inhibited the accumulation of triglyceride in the cytoplasm of adipocytes. (3) The mechanism that they inhibited C3H10T1/2 adipocyte differentiation is through decrease the level of PPARγ,C/EBPα,FABP4 mRNA and PPARγ,C/EBPα,FABP4 protein.