The Impact Of Puerarin On Proliferation、Apoptosis、Extracellular Matrix Of Human Embryo Fibroblasts And Its Potential Mechanism

Objective:To investigate the effects of puerarin on proliferation、apoptosis、extracellular matrix, and potential mechanism of action in TGF-β1-stimulated human embryo fibroblasts, which provide theoretical basis for the therapy of Chinese traditional and Western medicine.Method:200μg/ml puerarin was used for both TGF-p1stimulated human embryo lung fibroblasts and none stimulated. After24hours, with the different concentration we use the flow cytometer to mesure the apoptosis of HELFs and mesure proliferative activity by CCK-8, use Real Time PCR to mesure the changes of mRNA gene expression level for collagen-I and connective tissue growth factor(CTGF). Finally, Western blot used to mesure the changes of protein expression level for collagen-I and connective tissue growth factor.Result:The result of the measurement of flow cytometer shows in presence of24hs puerarin treatment could induce the apoptosis of HELFs in a dose-dependent manner with TGF-β1(P<0.01) stimulating. Puerarin, with concentration of100-200μg/ml, would inhibit cell proliferation in a dose-dependent manner(p<0.01), and seems much more obvious in TGF-β1-stimulated HELFs by CCK-8(P<0.01). By Real Time PCR, puerarin restrains the synthesis of CTGF and collagen-I mRAN of TGF-β1-stimulated HELFs in a dose-dependent manner(p<0.01). And as a result of western-blot, after5ng/ml TGF-β1stimulated, exprsession of a-SMA proteins in HELFs were significantly increased (P<0.05). Puerarin dose-dependently protein synthesis of CTGF and collagen-I in HELFs stimulated by TGF-β1(P<0.01).Conclusion:Puerarin could interfere IPF at the cellular level through affecting the synthesis of extracellular matrix collagen-I by inhibiting the proliferation of HELFs, inducing apoptosis and downregulating the expression of CTGF.