吡哆胺修复糖基化终产物诱导的脾淋巴细胞DNA损伤的实验研究

Determination of DNA damage induced by AGEs and DNA repair with pyridoxamine in mice spleen lymphocytes in vitro

目的 观察吡哆胺 (PM)对糖基化终产物 (AGEs)诱导的脾淋巴细胞DNA损伤的修复情况。方法 采用单细胞凝胶电泳 (SCGE)技术 ,根据AGEs BSA浓度的不同所致离体小鼠脾淋巴细胞的DNA损伤进行研究 ,并对PM修复AGEs BSA诱导的DNA损伤不同时点的修复能力作了分析。结果 ① 5 0、10 0 μg/mLAGEs BSA组脾淋巴细胞DNA迁移率与阴性对照组比较无显著性差异 (P >0 0 5 )。 2 0 0、4 0 0、80 0 μg/mLAGEs BSA组脾淋巴细胞DNA迁移率明显高于阴性对照组 (P <0 0 1) ,且两者呈正相关 (r=0 95 6 3)。②以能引起明显DNA断裂剂量 (2 0 0、4 0 0、80 0 μg/mL)的AGEs BSA作用小鼠脾淋巴细胞 ,PM最佳修复时点为 2h。结论 AGEs可引起离体脾淋巴细胞DNA链损伤 ,并存在一定的剂量 反应关系 ;

Objective To observe DNA damage induced by advanced glycation end-products(AGEs) and DNA repair with pyridoxamine(PM) in mice spleen lymphocytes in vitro. Methods The DNA damage effects of AGEs-BSA in different concentration were studied using the single cell gel electrophoresis(SCGE) technique in isolated mice spleen lymphocytes. So did the kinetics of DNA repair by PM in different time. Results We observed that in certain concentration (200, 400, 800 μg/mL), AGEs-BSA could induce the breakage of DNA single strand of mice spleen lymphocytes and result in comet cells with tail( P <0 01). The rate of DNA migration increased with concentration of AGEs-BSA. There were no significant difference between 50, 100 μg/mL AGEs-BSA group and control group( P >0 05). In addition, 200, 400, 800 μg/mL AGEs-BSA groups which induced DNA break were added to 10 mmol/L PM respectively. The optimal ratio of DNA repair was only seen within 2 hours during an incubation period of 1～4 hours after the operation. Conclusion Certain doses of AGEs-BSA could induce DNA damage of mice spleen lymphocytes in vitro and there is also an obvious dose-response-relationship between the rate of DNA migration and the dose of AGEs-BSA. Furthermore, the AGEs-BSA induced DNA damage was controlled by PM.