目的 考察天麻素对高糖诱导的人脐静脉内皮细胞（HUVEC）氧化应激的影响。方法 建立高糖诱导的HUVEC氧化损伤模型，用不同质量浓度的天麻素干预。MTT法检测HUVEC存活率；相关试剂盒测定一氧化碳（NO）的量、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）以及总抗氧化能力（T-AOC）的活性、丙二醛（MDA）的量，DCFH-DA染色法检测胞内活性氧（ROS）水平，实时荧光定量PCR法测定细胞中NF-κB 基因的表达。结果 与对照组相比，模型组HUVEC氧化应激作用明显增强，而天麻素能提高HUVEC中NO的量及SOD和CAT活性，提高细胞的总抗氧化能力，降低MDA的量与ROS水平，显著降低细胞中NF-κB 基因的表达。结论 天麻素通过提高抗氧化酶活性、降低脂质过氧化产物和ROS水平，抑制高糖诱导的HUVEC氧化应激作用，其作用机制可能与NF-κB通路密切相关。

Objective To establish a cell model for anti-oxidative drug screening by using anti-oxidative response element (ARE) to regulate the expression level of luciferase (Luc) reporter gene. Methods The recombinant plasmid vector pARE-Luc-Neo, expressed by ARE-regulated Luc reporter gene, was constructed. The recombinant plasmid vector was transfected into human embryonic kidney epithelial cells (Hek293 cells) using FuGENE^? HD transfection reagent. The positive cells were screened by G418, and the positive monoclonal cells were selected using the dilution method. The monoclonal cell lines with high activity of Luc reporter gene in response to anti-oxidative protein inducer TBHQ were obtained using the Luc activity detection. The screening effect of the positive clones was evaluated by resveratrol and curcumin. The effects of andrographolide, cryptotanshinone, silymarin, matrine, and polydatin at different concentration (0, 3.125, 6.25, 12.5, 25, 50, and 100 μmol/L) on Luc expression level were observed. Results The Hek293-ARE cells were obtained by detecting the Luc reporter gene activity, the Luc reporter gene expression was regulated by ARE, meanwhile there exists a dose-response relationship between the expression level and the concentration of the inducer within a certain range. The andrographolide, cryptotanshinone, and silymarin have significant induction effects. Conclusion The cell model for anti-oxidative drug screening is established successfully and could be effectively and high-throughput for the preliminary screening of anti-oxidative drugs.

国家高技术研究发展计划“863”项目（2011AA100603）；“重大新药创制”科技重大专项（2012ZX09102301）