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两种天然抗氧化剂对铁离子诱导的羟自由基形成以及细胞毒性的影响

Effects of Two Natural Antioxidants on Iron-Induced Hydroxyl Radical Production and Cytotoxity

【作者】 张丹

【导师】 张捷; 朱本占;

【作者基本信息】 华中农业大学 , 环境科学, 2009, 硕士

【摘要】 天然抗氧化物植酸和秦皮乙素用途广泛。本文以3-羧酸香豆素作为荧光探针研究了植酸、一磷酸腺苷(AMP)、三磷酸腺苷(ATP)、乙烯胺的乙酸衍生物EDTA、NTA和DTPA以及羟自由基清除剂如:二甲基亚砜(DMSO),甲酸(HCOOH)和乙醇(CH3CH2OH)对Fenton体系产生羟自由基的影响,同时选取HepG2细胞作为模型,探讨了秦皮乙素对铁参与的生物反应体系的影响,8HQ-Fe与细胞作用30分钟后加入秦皮乙素。研究结果表明:植酸可促使Fenton体系产生羟自由基(·OH);AMP和ATP对Fenton体系具有同样的促进作用;乙烯胺的乙酸衍生物EDTA、NTA和DTPA也能够促进·OH的产生;DMSO,HCOOH和CH3CH2OH可清除植酸-Fenton体系产生的·OH;同样,植酸可促使VO(Ⅱ)-H2O2反应(类Fenton体系)产生·OH;0.1 mM的8-羟基喹啉-铁(1:1)复合物对细胞有较大毒性;秦皮乙素本身对细胞不产生毒性,可保护细胞免受铁离子诱导的毒性,二者呈现一定的剂量-效应关系;秦皮乙素类似物4-甲基七叶苷原和秦皮素也具有相似的剂量-效应关系。其机理与铁的络合作用有关。该结果为进一步认识天然抗氧化物植酸和秦皮乙素提供参考。

【Abstract】 Phytic acid and esculetin has been widely used as antioxidant. In this study, coumarin-3-caroxylic acid was used as fluorescent probe to detect hydroxyl radical; meanwhile, the effect of esculetin on 8HQ-Fe-induced cell viability was investigated in HepG2 cells. Esculetin was added to the cells 30 min before the addition of 8HQ-Fe.The results indicated that: Phytic acid could markedly enhance·OH produced by ferrous iron and hydrogen peroxide; similar enhancing effects were also observed for AMP and ATP;·OH formation could be enhanced by aminoacetic chelating agents such as ethyleneamine ethylenediaminetetraacetic acid (EDTA), diethylenetriamine-penta acetic acid (DTPA) and nitrilotriacetic acid (NTA);·OH produced by Fe/ H2O2/phytic acid were markedly inhibited by·OH scanvenging agents such as dimethyl sulfoxide(DMSO), ethanol (CH3CH2OH) and formate(HCOOH); similar enhancing effect by phytic acid was observed in the reaction of vanadyl and H2O2 (Fenton-like reaction). Esculetin exhibited a dose dependent protection against iron-induced cell toxicity; similar dose-dependent protection was also observed with two other esculetin derivatives such as 6,7-dihydroxy-4-methylcoumarin and 7,8-dihydroxy-6-methoxycoumarin; in the absence of non-toxic esculetin but in the presence of concentrations of 8-hydroxyquinoline-Fe(II) complex (100μM 8-hydroxyquinoline, 100μM ferrous), cell toxicity was observed.The results are quite related to the mechanism of iron chelation, which help us know more about the property of these two antioxidants

  • 【分类号】X17
  • 【被引频次】1
  • 【下载频次】155
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