MODIFIED METHOD TO EVALUATE THE PROTECTION OF THE ANTIOXIDANTS AGAINST HYDROXYL RADICAL-MEDIATED DNA DAMAGE
- VernacularTitle:一种评价天然抗氧化剂对羟自由基致DNA损伤保护作用的改进方法
- Author:
Wei CAO
;
Weijun CHEN
;
Xiaohui ZHENG
;
Jianbin ZHENG
- Publication Type:Journal Article
- Keywords:
DNA damage;
hydroxyl radical;
Fenton reaction;
antioxidant
- From:
Acta Nutrimenta Sinica
1956;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a modified method to evaluate the protection of the antioxidant of DNA damage induced by hydroxyl radical. Method Reaction mixture at a final volume of 1.0 ml contained 0.8mg/ml DNA,0.2 mmol/L H2O2,1.0 mmol/L ascorbic acid,in 50 mmol/L NaH2PO4-Na2HPO4 buffer (pH 7.4). EDTA(1.0 mmol/L) and FeCl2 (1.0 mmol/L)were premixed and then dispensed into the reaction mixture to trigger the Fenton reaction,and the mixture was incubated at 37℃ for 30 min. The reaction was stopped by dispensing 1ml of 10% (w/v) trichloroacetic acid ,and reaction mixture was mixed with 1ml of 1% (w/v) 2-thiobarbituric acid,then further heated at 80 ℃ for 30 min. The chromogen formed was extracted into n-1-butanol and absorbance was measured at 532 nm. Results Addition of hydroxyl radical scavenger can compete with DNA for the hydroxyl radicals and diminished TBARS formation at a defined time of reaction with dose-dependent effect. The result showed that hydroxyl radical scavenger exerted inhibition effects on hydroxyl radical-mediated DNA damage. Conclusion This method can be used to evaluate the protection of the antioxidant of DNA damage induced by hydroxyl radical.
