龙胆苦苷与黄芩素对hiPS诱导的肝样细胞过氧化氢造成的氧化损伤的保护作用  

Protective Effect of Baicalein and Gentiopicroside on H_2O_2 Induced Liver Injury in Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells

作　　者：徐雯星 曾星[1] XU Wenxing;ZENG Xing(Department of Phase I Research Center,The Second Affiliated Hospital,GuangzhouUniversity of Chinese Medicine, Guangzhou 510006, Guangdong, China)

机构地区：[1]广州中医药大学第二临床医学院,广东广州510006

出　　处：《辽宁中医杂志》2019年第4期792-796,I0002,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：广东省中医院中医药科学技术研究专项(YK2013B2N10)

摘　　要：目的:建立H_2O_2致人诱导多能干细胞(i PSC)诱导的肝样细胞损伤模型并探讨龙胆苦苷与黄芩素对H_2O_2致肝样细胞损伤的保护作用。方法:采用四步诱导法,将人i PSC诱导为肝样细胞,将其分为正常组、模型组、中药单体干预组。除正常组外,其他组均以H_2O_2造模,建立肝样细胞氧化损伤模型,龙胆苦苷(gentiopicroside,GPS)做预防作用研究,在干预组中先加药,后造模。黄芩素(Baicalein,BAI)做保护作用研究,在干预组中先造模,后加药;通过收集细胞及细胞培养上清液,按照试剂盒的方法测定上清液中天冬氨酸转氨酶(ALT)、丙氨酸转氨酶(AST),细胞中丙二醛(MDA)的含量及超氧化物歧化酶(SOD)活性。结果:与正常组相比,模型组上清中ALT、AST升高,细胞中MDA含量显著升高,SOD活性显著降低,差异均具有统计学意义(P<0.01)。与模型组相比,黄芩素及龙胆苦苷干预组均可显著降低上清液中ALT和AST活性及肝样细胞中MDA含量,显著升高肝样细胞中SOD活性,差异均具有统计学意义(P<0.01)。结论:基于人i PSC诱导的肝样细胞成功建立H_2O_2氧化损伤模型,相较于其他肝损伤模型,此次造模的细胞是人源诱导而来,更贴近于人的病理,对肝损伤疾病具有临床指导意义。龙胆苦苷对H_2O_2致人诱导多能干细胞诱导的肝样细胞损伤有显著的预防作用。黄芩素对H_2O_2致人诱导多能干细胞诱导的肝样细胞损伤有显著保护作用。Objective:To establish a liver injury cell model by H2O2in human induced pluripotent stem cell(i PSC)-derived hepatocyte-like cells and explore the hepatoprotective effect of baicalein and gentiopicroside.Methods:Hepatocyte-like cells were produced by differentiation of human i PSC using an established 4-stage approach and then they were divided into normal group,model group and intervention group with different concentrations of baicalein and gentiopicroside.All the other groups were added H2O2to establish hepatocyte-like cell oxidation damage model except normal group.Subsequently intervention groups were exchanged with different concentrations of baicalein and gentiopicroside.The cells and their supernatants were collected.According to the method of the kit,the alanine transaminase(ALT)and aspartate transaminase(AST)in supernatant as well as malondialdehyde(MDA)and superoxide dismutase(SOD)activity in cells were measured.Results:Compared with normal group,supernatant levels of ALT and AST and cell MDA level were significantly increased in H2O2model group,along with a remarkable decrease in cell SOD activity and the differences were statistically significant(P<0.01).Compared with the model group,baicalein intervention group could significantly decrease supernatant ALT and AST levels and cell MDA content and increase cell SOD activity and the differences were statistically significant(P<0.01).Conclusion:The H2O2oxidative injury cell model was successfully established by human i PSC-derived hepatocyte-like cells.Compared with other models of liver injury,hepatocytelike cells in this model were induced from human species and more closer to human pathology.This may provide clinical significance for liver injury diseases.We conclude that baicalein and gentiopicroside have a significant protective effect on H2O2induced liver injury in human i PSC-derived hepatocyte-like cells.

关 键 词：黄芩素 龙胆苦苷 双氧水 肝样细胞 氧化损伤 肝保护 

分 类 号：R285[医药卫生—中药学]