芹菜素对H2O2诱导人肝细胞L02氧化损伤模型的影响

杜毅超; 张浩; 黄治伟; 浦仕林; 钱保林; 赖莉; 谭鹏; 夏先明; 付文广

doi:10.3969/j.issn.1001-5256.2020.05.025

芹菜素对H2O2诱导人肝细胞L02氧化损伤模型的影响

DOI: 10.3969/j.issn.1001-5256.2020.05.025

1. 西南医科大学附属医院四川省院士(专家)工作站
2. 西南医科大学附属医院肝胆外科

基金项目:

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）；

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中图分类号: R285.5
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出版历程
- 出版日期: 2020-05-20

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

Academician (Expert) Workstation of Sichuan Province, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, China

Research funding:

摘要

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<001）;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...
- 芹菜素 /
- 过氧化氢 /
- L02细胞 /
- 氧化损伤 /
- 细胞凋亡
Abstract:
Objective 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 de-hydrogenase(LDH), malondialdehyde(MDA), and superoxide dismutase(SOD) in cell supernatant; Hoechst staining was performed toobserve cell apoptosis, and a test kit was used to observe the activity of caspase-3. A one-way analysis of variance was used for compari-son of continuous data between multiple groups, and the LSD-ttest was used for further comparison between two groups.Results Apige-nin at a concentration of ≥20 μmol/L significantly inhibited the proliferation of L02 cells(P< 0. 01). Compared with the cells in the blankcontrol group, the cells treated with H2O2 at a concentration of ≥500 μmol/L had a significant reduction in cell viability(P< 0. 001), andtherefore, 500 μmol/L was determined as the optimal concentration for modeling. There was a significant difference in cell viability betweenthe model group and the blank control group(P< 0. 01), and compared with the model group, the 5 and 10 μmol/L apigenin groups had asignificant increase in cell viability(P< 0. 01). The cells in the blank control group had good morphology, while those in the model groupwere contracted and round-shaped and had marked rupture and deformity, and compared with the model group, the 5 μmol/L apigenin group showed significant improvement with a reduction in ruptured and round-shaped cells. There was a significant difference in fluores-cence intensity between the blank control group, the model group, and the 5 μmol/L apigenin group(1. 00 ± 0. 26 vs 32. 94 ± 1. 29 vs13. 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 5 μmol/L apigenin group had a significant reduction in H2O2-induced ROS(P<0. 001). Compared with the control group, the model group had significant increases in the levels of LDH and MDA and a significant reduc-tion in the level of SOD(F= 3. 21, 2. 03, and 3. 32, allP< 0. 05), and compared with the model group, the 5 μmol/L apigenin group hadsignificant reductions in the levels of LDH and MDA and a significant increase in the level of SOD(allP< 0. 05). There was a significantdifference in cell apoptosis rate between the blank control group, the model group, and the 5 μmol/L apigenin group(7. 54% ± 0. 52% vs39. 77% ± 3. 44% vs 14. 40% ± 0. 79%,F =9.439,P< 0. 01], and the model group had a significantly higher cell apoptosis rate than thecontrol group(P< 0. 01); the cells treated with apigenin had a significantly lower apoptosis rate than those in the model group(P< 0. 01).There was a significant difference in the activity of caspase-3 between the blank control group, the model group, and the 5 μmol/L apigeningroup(4. 38 ±0. 59 U/mg vs 16. 44 ±1. 13 U/mg vs 10. 60 ± 1. 04 U/mg,F =1.17,P< 0. 05), and the model group had a significantlyhigher activity of caspase-3 than the blank control group(P< 0. 05); compared with the cells in the model group, the cells treated withapigenin had a significant reduction in the activity of caspase-3(P< 0. 05).Conclusion Apigenin may exert a protective effect againstH2O2-induced injury in L02 cells by eliminating ROS and reducing caspase-3 activity.
- apigenin /
- hydrogen peroxide /
- LO2 cells /
- oxidative damage /
- apoptosis

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