Webb Michael R, Min Kyungmi, Ebeler Susan E
Department of Viticulture and Enology, University of California-Davis, One Shields Avenue, Davis, CA 95616, U.S.A.
J Food Biochem. 2008 Sep 23;32(5):576-596. doi: 10.1111/j.1745-4514.2008.00181.x.
Anthocyanins and their aglycone anthocyanidins are pigmented flavonoids found in significant amounts in many commonly consumed foods. They exhibit a complex chemistry in aqueous solution, which makes it difficult to study their chemistry under physiological conditions. Here we used a gel electrophoresis assay employing supercoiled DNA plasmid to examine the ability of these compounds (1) to intercalate DNA, (2) to inhibit human topoisomerase I through both inhibition of plasmid relaxation activity (catalytic inhibition) and stabilization of the cleavable DNA-topoisomerase complex (poisoning), and (3) to inhibit or enhance oxidative single-strand DNA nicking. We found no evidence of DNA intercalation by anthocyan(id)ins in the physiological pH range for any of the compounds used in this study-cyanidin chloride, cyanidin 3-O-glucoside, cyanidin 3,5-O-diglucoside, malvidin 3-O-glucoside and luteolinidin chloride. The anthocyanins inhibited topoisomerase relaxation activity only at high concentrations (> 50 muM) and we could find no evidence of topoisomerase I cleavable complex stabilization by these compounds. However, we observed that all of the anthocyan(id)ins used in this study were capable of inducing significant oxidative DNA strand cleavage (nicking) in the presence of 1 mM DTT (dithiothreitol), while the free radical scavenger, DMSO, at concentrations typically used in similar studies, completely inhibited DNA nicking. Finally, we propose a mechanism to explain the anthocyan(id)in induced oxidative DNA cleavage observed under our experimental conditions.
花青素及其糖苷配基花色素是在许多常见食用食物中大量存在的色素类黄酮。它们在水溶液中呈现出复杂的化学性质,这使得在生理条件下研究其化学性质变得困难。在此,我们使用了一种采用超螺旋DNA质粒的凝胶电泳分析方法,来检测这些化合物的能力:(1)嵌入DNA;(2)通过抑制质粒松弛活性(催化抑制)和稳定可裂解的DNA-拓扑异构酶复合物(中毒)来抑制人拓扑异构酶I;以及(3)抑制或增强氧化性单链DNA切口。我们发现,在本研究中使用的任何化合物——氯化花青素、花青素3-O-葡萄糖苷、花青素3,5-O-二葡萄糖苷、锦葵色素3-O-葡萄糖苷和氯化木犀草素——在生理pH范围内均无花青素(苷元)嵌入DNA的证据。花青素仅在高浓度(>50μM)时抑制拓扑异构酶松弛活性,且我们未发现这些化合物稳定拓扑异构酶I可裂解复合物的证据。然而,我们观察到,在本研究中使用的所有花青素(苷元)在存在1 mM二硫苏糖醇(DTT)的情况下均能够诱导显著的氧化性DNA链断裂(切口),而在类似研究中通常使用的浓度下,自由基清除剂二甲基亚砜(DMSO)完全抑制了DNA切口。最后,我们提出了一种机制来解释在我们的实验条件下观察到的花青素(苷元)诱导的氧化性DNA裂解。