砷对人淋巴细胞 DNA 氧化损伤的作用

目的：探讨砷（As)引起植物血凝集素（PHA）刺激和无刺激人外周血淋巴细胞DNA氧化性损伤。方法：用10μmol/L砷处理细胞2h,经单细胞凝胶电泳（SCGE,或彗星试验）－FPG（甲酰胺基嘧啶－DNA糖基化酶）消化法检测砷引起的DNA碱基损伤。结果：砷引起的DNA链断裂的修复过程与过氧化氢（H2O2）引起的修复过程类似,FPG消化产生的单链断裂,或砷引起的碱基损伤在PHA刺激淋巴细胞较未刺激细胞显著,在PHA刺激的淋巴细胞,砷和H2O2引起的DNA链断裂2h分别修复63％和68％,但在未刺激细胞分别修复大约34％和43％,在PHA刺激的淋巴细胞,砷和H2O2引起的碱基损伤2h分别修复40％和49％,但在未刺激细胞分别修复大约19％和21％。结果：微量砷可引起人类细胞DNA氧化性损伤,损伤的碱基主要是嘌呤或甲酰胺基嘧啶,未分裂（刺激）淋巴细胞修复砷与H2O2引起的DNA损伤较慢。

Application of SCGE-FPG in the study of arsenic-induced oxidative DNA damage in PHA-stimulated and unstimulated human lymphocytes

OBJECTIVE: To confirm that arsenic (As) induces oxidative DNA damage in phytohemagglutinin (PHA)-stimulated and unstimulated human lymphocytes. METHODS: The alkaline comet assay combined with specific enzyme (Formamidopyrimidine-DNA glycosylase, FPG) digestion was used to measure As-induced base damage. RESULTS: The enzyme-sensitive sites were readily detected with the alkaline comet assay after the cells were treated with 10 micromol As for 2 hours. The repair patterns observed for FPG-created DNA single strand breaks (SSBs) in As-treated cells were comparable to those in hydrogen peroxide (H(2)O(2))-treated cells. The enzyme-created SSBs, As-induced base damage, were more significantly revealed in PHA-stimulated lymphocytes. About 63% and 68% of SSBs induced by As and H(2)O(2), respectively, were repaired in PHA-stimulated lymphocytes by 2-hour repair incubation, but about 34% and 43%, respectively, were repaired in unstimulated cells. About 40% and 49% of base damage induced by As and H(2)O(2), respectively, were repaired in PHA-stimulated lymphocytes, but about 19% and 21 %, respectively, were repaired in unstimulated cells. CONCLUSIONS: As induces oxidative DNA damage in human lymphocytes within micromolar concentrations. Like the damage induced by H(2)O(2), As-induced DNA damage was more slowly repaired in unstimulated lymphocytes.