石榴皮多酚纯化物对LO2肝细胞的损伤影响

Effects of purified pomegranate peel polyphenols on LO2 hepatocytes

目的探究石榴皮多酚纯化物(安石榴苷含量高于80%)对LO2肝细胞的损伤影响。方法将LO2肝细胞分为药物组(给予3.125、6.250、12.500、25.000、50.000、100.000、20.000μg/mL的石榴皮多酚纯化物)、阴性组(给予正常培养基),置于37℃、5%CO2环境下培养24h后,采用CCK8法测定OD值,计算抑制率;采用Elisa法检测细胞培养液中天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、丙氨酸氨基转移酶(ALT)释放量的变化;采用Hoechst荧光染色法观察LO2肝细胞的形态变化和凋亡情况。结果与阴性组比较,不同浓度药物组OD值降低,LO2肝细胞存活率降低,差异有统计学意义(P <0.01);不同浓度药物组细胞培养液中ALT、AST、ALP含量降低,差异有统计学意义(P <0.05)。结论石榴皮多酚纯化物对体外培养LO2肝细胞无损伤作用。

Objective To explore the effects of purified pomegranate peel polyphenols on LO2 hepatocytes.Methods The LO2 hepatocytes were divided into a series of drug concentration-gradient groups(3.125,6.250,12.500,25.000,50.000,100.000,20.000μg/mL pomegranate polyphenol purified),and a negative group(administered with normal medium),were placed at 37℃,5%CO-2 environment for 24 h.The OD value was measured by the CCK8 method,and the inhibition rate was calculated.The ELISA method was used to detect the changes of aspartate aminotransferase(AST),alkaline phosphatase(ALP)and alanine aminotransferase(ALT)release in cell culture medium.Hoechst fluorescence staining was used to observe the morphological changes and apoptosis of LO2 hepatocytes.Results Compared with the negative group,the OD values of the different concentrations of the drug groups decreased,and the survival rate of the liver cells of LO2 decreased,which the difference was statistically significant(P<0.01).Compared with the negative group,the level of ALT,AST and ALP in the cell culture medium was decreased,and the difference was statistically significant(P<0.05).Conclusion The purified pomegranate peel polyphenol has no damage to cultured LO2 hepatocytes in vitro.