5'-磷酸腺苷对小鼠脾细胞氧化应激损伤的保护作用

PROTECTIVE ROLE OF 5'-ADENYLIC ACID AGAINST OXIDATIVE STRESS INJURY OF MOUSE SPLENIC CELLS

目的研究5'-磷酸腺苷(5'-AMP)体外抗氧化和对体外氧化损伤脾细胞的损伤修复能力。方法用化学比色法测定5'-AMP体外清除二苯代苦味酰基自由基(DPPH自由基)的能力;建立过氧化氢(H2O2)氧化损伤体外培养小鼠脾细胞模型,用MTT法检测5'-AMP修复受损伤脾细胞的作用,并分析其对细胞抗氧化体系及抗氧化能力的影响。结果5'-AMP具有剂量依赖性的体外抗氧化和清除活性氧能力,添加0·5mmol/L、1mmol/L、5mmol/L和10mmol/L5'-AMP均能显著修复H2O2诱导的脾细胞氧化损伤(P<0·05),总抗氧化能力和抗氧化酶类活力(P<0·01),5'-AMP添加量大于1mmol/L时,可显著降低丙二醛(MDA)含量(P<0·01)。其细胞培养液的氧自由基(ROS)水平逐渐降低,5'-AMP添加量为10mmol/L时,ROS水平接近对照组水平。结论5'-磷酸腺苷能显著修复氧化损伤,具有显著的抗氧化作用。

Objective To investigate in vitro antioxidation of 5′-adenylic acid （5′-AMP） and its repairing effect on oxidative damage in spleen cells. Methods Scavenging of DPPH free radicals by 5′-AMP was studied by chemical colorimetry. Mouse splenic cells were treated with H2O2 to induce oxidative stress injury, which was determined. The effect of 5′-AMP on injury was studied by MTT method. Total antioxidation capability （T-AOC）, catalase （CAT） activities, glutathione peroxidase （ GSH-Px ） activities, diaminodiphenyl methane （MDA） and reactive oxygen species （ROS） level in the incubation medium were detected. Results 5′-AMP could scavenge DPPH radicals in a concentration dependent manner in vitro and significantly repair the injury by H2O2 in splenic cells. Compared to that of the untreated group, the number of living cells increased significantly in H2O2-injured splenic cells with the supplementation of 0.5, 1, 5 and 10mmol/L 5′-AMP （P〈0.05） . CAT activities, GSH-Px activities and T-AOC in the incubation medium were increased significantly with 5′-AMP concentration increase （P〈0.01）, while MDA content decreased significantly （P〈0.01） with the supplementation of more than 1mmol/L 52AMP. ROS level decreased gradually, and 10mmol/L of 5′-AMP obtained the best effect, which was close to the control group. Conclusion 5′-AMP has a significant anti-oxidation activity, which can significantly repair the oxidative injury.