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目的：研究不同浓度葡萄糖氧化酶（GO）对人肝细胞L02 氧化应激水平的影响，以确定建立肝细胞氧化应激模型的合适浓度。方法：用不同浓度GO干预L02 肝细胞2 h，MTT法检测细胞的存活率，流式细胞术检测细胞内活性氧簇(ROS)，荧光强度（FI）来表示ROS水平。分光光度法检测检测细胞MDA、GSH，速率法检测细胞培养液LDH、AST和ALT的水平。结果：① 随GO浓度增加，肝细胞的存活率逐渐降低，其中75 U/L、100U/L和125 U/L组存活率显著低于对照组（P<0.05）。②随GO 浓度增加，MDA 含量逐渐增高，其中50 U/L、75 U/L、100 U/L、125 U/L组MDA水平较对照组显著增高（P<0.05）。GSH水平随GO 浓度增高而逐渐减低，各干预组较对照组均显著降低（P<0.05）。GO各干预组FI均较对照组显著降低（P<0.05）。③各干预组LDH活性均显著高于对照组（P<0.05），50 U/L、75 U/L、100 U/L、125 U/L干预组AST与ALT水平均较对照组显著增高（P<0.05）。结论：GO能引起的肝细胞氧化应激损伤有剂量依赖性，100 U/L是建立肝细胞氧化应激的合适浓度。

英文摘要:

Objective: To investigate the effect of glucose oxidase(GO) on oxidative stress to establish a model for oxidative stress in cultured human LO2 hepatocytes. Methods: LO2 hepatocytes were treated with different concentrations of GO. The viability of LO2 hepatocyte was evaluated by MTT assay. The intercellular ROS level was analyzed on a flow cytometer by dihydroethidium and was indicated with fluorescence intensity (FI). The levels of MDA and GSH were measured by spectrophotometric method. The levels of LDH, AST and ALT were measured by kinetic rate method using commercially available reagent kits. Results: ①The viability of LO2 hepatocytes decreased along with the increase in concentration of GO. The viabilities of hepatocytes in GO group 75 U/L, 100U/L and 125 U/L were significantly higher than those in control group (P<0.05). ②The levels of MDA increased along with the increase in concentration of GO, MDA levels in GO group 75 U/L, 100U/L and 125 U/L significantly were higher than those in control group (P<0.05). Compared with control group, the levels of GSH decreased significantly in different concentrations of GO groups (P<0.05). The levels of FI increased significantly in different concentrations of GO groups compared with those in the control group (P<0.05). ③The LDH levels increased significantly in model groups treated with different concentrations of GO compared with those in control group (P<0.05). The levels of AST and ALT increased with the increase in concentration of GO, the levels in different concentration of GO groups were significantly higher in comparison with control group (P<0.05). Conclusion: Oxidative stress induced by GO is dose-dependent in cultured hepatocyte L02, and 100 U/L maybe the ideal concentration to establish the hepacytoe model with oxidative stress.

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