Effects Of Grape Procyanidins On The Concentration Of Intracellular Calcium In Hepatocytes

OBJECTIVETo investigate the effects of grape procyanidins on the concentration of intracellular calcium in hepatocytes. METHODS 1. Vitro ExperimentsWe cultured 4 types of different hepatocytes,that is, normal hepatocytes, hepatocytes treated with 1mmol/L H2O2 or 100mg/L ethanol and hepatoma cells.The microfluorimetric techniques based on the calcium indicator Fura-2/AM were utilized to investigate intracellular calcium concentration and MTT assay was utilized to investigate the proliferation of different hepatocytes treated with GPC. 2.Vivo Experiments30 mice were randomly divided into 5 groups,normal group,GPC control group,ethanol control group,low-dosage GPC group and high-dosage GPC group. The mice in normal group were gavaged only with water.The mice in GPC control group and high-dosage GPC group were gavaged with 100mg/kg body weight. The mice in low-dosage GPC group were gavaged with 1 Omg/kg body weight. Furthmore ,on the fifth and sixth days,the mice in ethanol control group,low-dosage GPC group and high-dosage GPC group were gavaged with ethanol 4000mg/kg body weight.On the seventh day,Fura-2/AM technique was utilized to investigate intracellular calcium concentration, MTT assay was utilized to investigate the proliferation and immunohistochemistry method and imaging analysis technique was applied to measure the expression level of Bcl-2 and Bax of hepatocytes in every group. RESULTSThe concentration of intracellular calcium and proliferation were 108.26 +14.17 nmol/L and 0.320+0.016 respectively in vitro normal hepatocytes.But the two indexes eleveted to 149.44 ± 11.18nmol/L and 0. 515 ±0. 013 in 50mg/L GPC group. Meanwhile they eleveted to176.46 + 19.36nmol/L 和 0. 756+0. 045 in 100mg/L GPC group. Furthmore,There are significant diferences between the three groups(P<0.05).