Suppression of oxidative cellular damage by quinone reductase-inducible constituents in Vegetables of Umbelliferae

伞形科蔬菜中醌还原酶诱导成分对氧化细胞损伤的抑制

• 批准号: 13660116
• 负责人: UDA Yasushi
• 金额: $ 1.6万
• 依托单位: Utsunomiya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660116/
• 关键词: Quinone reductase; HepG2; Ashitaba; Umbelliferae; Chalcone derivative; キノンレダクターゼ誘導; 8-OH-デオキシグアノシン; あしたば

Isolation of quinone reductase (QR)-inducible constituents from freeze-dried Ashitaba leaves (Angelica keiskei Koidz.) was performed. The dried leaves were extracted with 80 % methanol, and ethyl acetate-soluble fraction was separated therein. The fraction was then chromatographed on a silica gel column by eluting with mixtures of n-hexane and ethyl acetate, in which six major components (Pk1, Pk2, Pk3, Pk4, Pk5, and Pk6) were detected by HPLC analysis. Among the components, both Pk3 and Pk4 were found to induce QR. In addition, the component, Pk6 showed a strong cytotoxicity. The QR-inducible components, Pk3 and Pk4, were further purified using a C18 Sep-Pak cartridge, by eluting the cartridge with 50% and 70 % acetonitrile in water, respectively. Also, the cytotoxic component, Pk6 was isolated on the cartridge by eluting with 100% acetonitrile. These isolates were analyzed by HPLC-MS. Mass spectrum of the QR-inducer, Pk3 showed an apparent fragment ion at m/z 285 which may be due to a flavonol, luteolin, along with the molecular ion peak at 645 (M^+-H). It was presumed that this compound might be a flavonol glycoside having luteolin as aglycone. On the other hand, the cytotoxic component showed an ion peak at m/z 338 as the molecular ion, which was identical to that of a chalcone derivative, 4-hydroxy derrick (2',4-dihydroxy-3'-(γ,γ-dimethylallyl)-4'- methoxychalcone). This suggested that the cytotoxic component should be 4-hydroxy derricin, which has been known as a constituent of Ashitaba root. The component was found in other vegetables such as Mitsuba (Cryptotaenia japonica Hassk), Seri (Oenanthe javanica DC.), and Parsley (Petroselium crispum Nym.). Effect of the components, Pk3 and Pk4 on oxidative cellular damage was evaluated on the basis of amount of 8-OhdG formed by menadione-mediated oxidation of DNA, in which both components showed no suppressive effect. This should be further studied along with improvement of the evaluation method.

冻干当归(Angelica keiskei Koidz.)中醌还原酶(QR)诱导成分的分离已经被执行了。干叶用80%甲醇提取，分离出乙酸乙酯可溶物。用正己烷和乙酸乙酯混合溶剂洗脱，在硅胶柱上进行层析，用高效液相色谱分析了6种主要成分(Pk1、Pk2、Pk3、Pk4、Pk5和Pk6)。在这些成分中，Pk3和PK4都能诱导QR。此外，该成分Pk6具有较强的细胞毒作用。QR诱导成分Pk3和PK4用C18Sep-Pak柱进一步纯化，分别用50%和70%的乙腈在水中洗脱。此外，用100%乙腈洗脱，在小柱上分离出细胞毒性成分Pk6。对分离的菌株进行了高效液相色谱-质谱仪分析。QR诱导剂Pk3的质谱图显示在m/z 285处有明显的碎片离子，这可能是由于黄酮醇，木犀草素，以及在645(M^+-H)处的分子离子峰。推测该化合物可能是以木犀草素为苷元的黄酮苷。另一方面，细胞毒组分的分子离子在m/z338处出现一个离子峰，与查尔酮衍生物4-羟基-3‘-(γ，γ-二甲基烯丙基)-4’-甲氧基查尔酮相同。这表明细胞毒性成分应该是4-羟基地黄毒素，它已被认为是灰树根中的一种成分。在其他蔬菜中也发现了这种成分，如Mitsuba(Cryptotaenia Japan Onica Hassk)、Seri(Oenanthe javanica DC.)和欧芹(Petroselum crispum Nym.)。根据甲萘二酮介导的DNA氧化生成8-OHdG的量来评价这两个组分对细胞氧化损伤的影响，其中两个组分都没有抑制作用。这需要随着评价方法的改进而进一步研究。

项目成果

Kei Hashimoto: "In vitro induction of the anticarcinogenic marker enzyme, quinone reductase, in human hepatoma cells by food extracts"Cancer Letters. 180. 1-5 (2002)

Kei Hashimoto：“通过食物提取物在人肝癌细胞中体外诱导抗癌标记酶醌还原酶”《癌症快报》。

Kei Hashimoto, Shinsuke Kawamata, Nomi Usui, Ayako Tanaka, and Yasushi Uda: "In vitro induction of the anticarcinogenic marker Enzyme, in human hepatoma cells by food extract"Cancer Letters. 180. 1-5 (2002)

Kei Hashimoto、Shinsuke Kawamata、Nomi Usui、Ayako Tanaka 和 Yasushi Uda：“通过食物提取物在人肝癌细胞中体外诱导抗癌标记酶”《癌症快报》。

Kei Hashimoto: "In vitro induction of the anticarcinogenic marker enzyme, quinone reductase, in human hepatoma cells by food extracts"Cancer Letters. Vol.180. 1-5 (2002)

Kei Hashimoto：“通过食物提取物在人肝癌细胞中体外诱导抗癌标记酶醌还原酶”《癌症快报》。