铜致L-02细胞的氧化应激及凋亡的影响

Oxidative Damage on Human Liver Cells L-02 by Copper

[目的]研究铜致人肝细胞L-02的氧化应激及凋亡的影响。[方法]噻唑蓝(MTT)法检测细胞生长活性。黄嘌呤氧化酶法检测细胞培养上清液中超氧化物歧化酶(SOD)活力。比色法测定还原型谷胱甘肽(GSH)含量。硫代巴比妥酸法(TBA)检测氧化产物丙二醛(MDA)含量。流式细胞术(FCM)检测细胞凋亡率。[结果]铜呈剂量依赖性和时间依赖性地抑制细胞生长活性,50、100、150、200μmol/L浓度组细胞生长抑制率分别为6.27%、16.89%、30.64%和39.32%,6、12、18、24h组分别为7.63%、16.83%、33.66%和38.74%,与对照组比较差异有统计学意义(P<0.05,P<0.01)。铜抑制细胞培养上清液中抗氧化酶SOD活性,降低GSH含量,使氧化产物MDA生成增加,各浓度组与与对照组比较差异有统计学意义(P<0.05,P<0.01)。FCM结果表明随着铜浓度增大和时间延长,细胞凋亡率逐渐上升。5、100、150、200μmol/L浓度组细胞凋亡率分别为4.06%、8.39%、13.81%和17.07%,6、12、18、24h组分别为5.30%、7.39%、13.81%和20.45%.与对照组比较差异有统计学意义(P<0.05,P<0.01)。[结论]铜可致人肝细胞L-02氧化应激,抑制细胞生长,诱导细胞凋亡。

[ Objective ] To study the oxidative damage on human liver cells L-02 by copper. [ Methods ] The viability of cells was determined by 3-[ 4, 5-dimedthyhhiazol ]-2, 5-diphenyltetrazolium bromide（ MTT ） assay. The activity of superoxide dismutase （ SOD ）was detected by xanthine oxidase method, and the content of Glatathiane （ GSH ）was measured by colorimetric and malondialdehyde （ MDA ）was determinated by thiobituric acid reaction（ TBA ）method in culture supernatant fluid. The changes of the cell apoptosis was detected by flow cytometer. [ Results ] Copper could inhibit the viability of cells in concentration and time dependent. The inhibitory rate increased from 6.27% to 39.32% as the dosage increased from 50 to 200 μmol/L, and from 7.63% to 38.74% as the time increased from 6 to 24h, which were significantly higher than that of the control（ P〈0,05, P〈 0.01 respectively ）.Copper could restrain the activity of SOD, and decrease the content of GSH and increase the level of MDA in culture supernatant fluid. The result of flow cytometer indicated that cell apoptosis increased in concentration and time dependent. The rate of apoptosis increased from 4.06% tel 7.07% as the dosage increased from 50 to 200μmol/L, and from 5.30% to 20.45% as the time increased from 6 to 24h, which were significantly higher than that of the control （P〈0.05, P〈0.01 respectively ）. [ Conclusion ] Copper can induce oxidative damage on human liver cell L-02, and can inhibit cell＇ s growth and result in apoptosis