芹菜素对H2O2诱导人肝细胞L02氧化损伤模型的影响  

Effect of apigenin on H2O2-induced oxidative injury in human hepatocytes L02

作　　者：杜毅超 张浩 黄治伟 浦仕林 钱保林 赖莉 谭鹏 夏先明 付文广 DU Yichao;ZHANG Hao;HUANG Zhiwei(Academician (Expert) Workstation of Sichuan Province, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, China)

机构地区：[1]西南医科大学附属医院四川省院士(专家)工作站,四川泸州646000 [2]西南医科大学附属医院肝胆外科,四川泸州646000

出　　处：《临床肝胆病杂志》2020年第5期1077-1081,共5页Journal of Clinical Hepatology

基　　金：四川省应用基础研究基金(2018JY0283);四川省定向财力转移支付科技项目(2017SZYZF0015)。

摘　　要：目的探索芹菜素对H2O2致人肝细胞L02损伤模型的保护作用。方法以H2O2诱导L02细胞建立氧化损伤模型,CCK-8法检测细胞存活率,DCFH-DA法检测细胞活性氧的生成,试剂盒检测细胞上清液中乳酸脱氢酶(LDH)、丙二醛(MDA)及超氧化物歧化酶(SOD)活性,Hoechst染色观察细胞凋亡情况,试剂盒检测caspase-3的活性。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果芹菜素浓度为≥20μmol/L时对L02细胞的增殖有显著抑制作用(P值均<0.01);与空白对照组L02细胞活力相比,500μmol/L及以上的H2O2浓度均可使细胞活力极显著降低(P值均<0.001),500μmol/L为H2O2的建模浓度;模型组细胞活力与空白对照组相比差异显著(P<0.01),与模型组相比,芹菜素5、10μmol/L组细胞存活率显著提高(P值均<0.01);空白对照组细胞状态较好,模型组细胞间收缩变圆,细胞破损变形严重,芹菜素5μmol/L组与模型组相比,明显得到改善,变圆破损细胞较少;空白对照组、模型组、芹菜素5μmol/L组3组间相对荧光强度比较差异有统计学意义(1.00±0.26 vs 32.94±1.29 vs 13.49±1.23,F=1.10,P<0.001),模型组相对荧光强度较空白对照组明显增强(P<0.001),芹菜素5μmol/L组与模型组比较,芹菜素可明显清除H2O2诱发的ROS(P<0.001);模型组中LDH和MDA水平均明显升高,SOD水平明显降低,与正常对照组相比差异均有统计学意义(F值分别为3.21、2.03、3.32,P值均<0.05),芹菜素(5μmol/L)处理后,与模型组相比,LDH和MDA水平均明显降低,SOD水平明显升高(P值均<0.05);空白对照组、模型组、芹菜素5μmol/L组3组间细胞凋亡率比较差异有统计学意义(7.54%±0.52%vs 39.77%±3.44%vs 14.40%±0.79%,F=9.44,P<0.01);模型组细胞凋亡率显著高于对照组(P<0.01);给与芹菜素处理后,相比于模型组凋亡率明显降低(P<0.01);空白对照组、模型组、芹菜素5μmol/L组3组间caspase-3活性比较差异有统计学意义[(4.3Objective To investigate the protective effect of apigenin against H2O2-induced injury in human hepatocytes(L02).Methods L02 cells were treated with H2O2 to establish a model of oxidative injury.CCK-8 assay was used to measure cell viability;DCFH-DA was used to measure the production of reactive oxygen species(ROS)in cells;test kits were used to measure the activities of lactate dehydrogenase(LDH),malondialdehyde(MDA),and superoxide dismutase(SOD)in cell supernatant;Hoechst staining was performed to observe cell apoptosis,and a test kit was used to observe the activity of caspase-3.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the LSD-t test was used for further comparison between two groups.Results Apigenin at a concentration of≥20μmol/L significantly inhibited the proliferation of L02 cells(P<0.01).Compared with the cells in the blank control group,the cells treated with H2O2 at a concentration of≥500μmol/L had a significant reduction in cell viability(P<0.001),and therefore,500μmol/L was determined as the optimal concentration for modeling.There was a significant difference in cell viability between the model group and the blank control group(P<0.01),and compared with the model group,the 5 and 10μmol/L apigenin groups had a significant increase in cell viability(P<0.01).The cells in the blank control group had good morphology,while those in the model group were contracted and round-shaped and had marked rupture and deformity,and compared with the model group,the 5μmol/L apigenin group showed significant improvement with a reduction in ruptured and round-shaped cells.There was a significant difference in fluorescence intensity between the blank control group,the model group,and the 5μmol/L apigenin group(1.00±0.26 vs 32.94±1.29 vs 13.49±1.23,F=1.10,P<0.001),and the model group had a significantly higher fluorescence intensity than the blank control group(P<0.001).Compared with the model group,the 5μmol/L apigenin group had a significant reduc

关 键 词：芹菜素 过氧化氢 L02细胞 氧化损伤 细胞凋亡 

分 类 号：R575[医药卫生—消化系统]