Flavonoid inhibition of the human neutrophil NADPH-oxidase.
Flavonoid inhibition of the human neutrophil NADPH-oxidase.
复制标题
类黄酮抑制人中性粒细胞 NADPH 氧化酶。
DOI：
10.1016/0006-2952(84)90194-1
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发表时间：
1984
影响因子：
5.8
通讯作者：
Marletta,MA
中科院分区：
文献类型：
--
作者：
Tauber,AI;Fay,JR;Marletta,MA
材料与方法槲皮素、山奈酚、桑色素、还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）、酵母聚糖、细胞色素c（VI型）、乳酸脱氢酶试剂盒、超氧化物歧化酶（SOD）和佛波醇12-肉豆蔻酸酯，13-乙酸酯（PMA）购自Sigma Chemical Co. (St. Louis，MO），并且非瑟酮从Aldrich Chemical Co.（密尔沃基，WI）。通过HPLC测定黄酮醇的最终纯度。HPLC系统由反相柱（来自沃茨Associates，Inc.的uBondapak C18）和线性甲醇/水（1%乙酸）梯度，在20分钟内从45%甲醇至100%。样品以50%甲醇进样，黄酮醇的储备液用二甲基亚砜（DMSO）配制，任何实验中DMSO的终浓度均不超过1%（v/v）。通过Hansch及其同事[9]报告的方法测定黄酮醇的分配系数，不同之处在于水溶液部分由细胞实验中使用的磷酸盐缓冲盐水缓冲液组成。将两相体系通过涡旋混合1分钟并使其分离，然后测量水相在370 nm处的吸光度。
Materials and methodsQuercetin, kaempferol, morin, nicotinamide adenine dinucleotide phosphate, reduced form (NADPH), zymosan, cytochrome c (Type VI), lactate dehydrogenase kit, superoxide dismutase (SOD) and phorbol12-myristate, 13-acetate (PMA) were purchased from the Sigma Chemical Co.(St. Louis, MO), and fisetin was uurchased from the Aldrich Chemical Co.(Milwaukee, WI). The final purity of the flavonols was determined by HPLC. The HPLC system consisted of a reverse-phase column (, uBondapak Cl8 from Waters Associates, Inc.) and a linear methanol/water (1% acetic acid) gradient from 45% methanol to 100% over 20min. The samples were injected at 50% methanol.The stock solutions of flavonols were made up in dimethyl sulfoxide (DMSO), and the final concentration of DMSO in any experiment did not exceed 1%(v/v). The partition coefficient of the flavonols was determined by the method reported by Hansch and colleagues [9] except that the aqueous portion consisted of the phosphate-buffered saline buffer used in the experiments with the cells. The twophase system was mixed by vortex for 1 min and allowed to separate, and then the absorbance at 370nm of the aqueous phase was measured.