权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Studies on the inhibitors of the catalytic activities and gene expressions of arachidonate oxygenases

花生四烯酸加氧酶催化活性及基因表达抑制剂的研究

基本信息

批准号：
14570113
负责人：
YAMAMOTO Shozo
金额：
$ 2.18万
依托单位：
Kyoto Women's University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

The biosynthesis of lipid mediators from arachidonic acid is initiated by cyclooxygenase and lipoxygenase enzymes. The purpose of this research project is to search for inhibitors of the catalytic activities of these enzymes and the transcription of enzyme genes.1)Transcription inhibitor of cyclooxygenase gene : Earlier we reported that humulone isolated from beer hop inhibited the transcription of cyclooxygenase-2 gene. In connection of the role of cyclooxygenase-2 in angiogenesis, we found that the angiogenesis of chicken embryo chorioallantoic membrane was markedly inhibited by humulone.2)12-Lipoxygenase inhibitor : Earlier we found that hinokitiol, a tropolon derivative, was a selective inhibitor of the platelet-type among the three isozymes of 12-lipoxygenase ; platelet-, leukocyte- and epidermis-types. Further studies demonstrated a higher specificity of the inhibition of platelet 12-lipoxygenase with an ether of hinokitiol and cinnamic alcohol. Furthermore, we noted the antioxidant activity of tea leaf catechins, and tested them as an inhibitor of 12-lipoxygenase. Most catechins tested inhibited the enzyme of platelet-type relatively selectively. The most potent and selective inhibition was observed with (-)-gallocatechin gallate with an IC50 of 0.1μM. In addition, hinokitiol at about 10μM inhibited the growth of various cell lines of melanoma and prostate cancer3)Enzyme immunoassay of prostaglandin E2 : Among various cyclooxygenase-metabolites of arachidonic acid, prostaglandin E2 shows a variety of biological activities, and its production is assayed to follow the cyclooxygenase expression. As an easy and sensitive assay, we attempted to develop an enzyme immunoassay of prostaglandin E2. We developed an assay with acetylcholine esterase as a label which gave a calibration curve with an IC50 of 4.5 pg/50 μl sample.

由花生四烯酸生物合成脂质介质是由环氧合酶和脂氧合酶启动的。本研究的目的是寻找这些酶的催化活性和酶基因转录的抑制剂。1）环氧合酶基因的转录抑制剂：早期我们报道了从啤酒花中分离的蛇麻酮抑制环氧合酶-2基因的转录。2）12-脂氧合酶抑制剂：在12-脂氧合酶的三种同工酶（血小板型、白细胞型和表皮型）中，我们发现一种托酚衍生物扁柏酚是血小板型的选择性抑制剂。进一步的研究表明，扁柏酚和肉桂醇的醚对血小板12-脂氧合酶的抑制具有更高的特异性。此外，我们注意到茶叶儿茶素的抗氧化活性，并测试它们作为12-脂氧合酶的抑制剂。大多数儿茶素对血小板型酶有较强的抑制作用。在（-）-没食子儿茶素没食子酸酯中观察到最有效和选择性的抑制作用，IC 50为0.1μM。此外，约10μM的扁柏酚抑制各种黑色素瘤和前列腺癌细胞系的生长3）前列腺素E2的酶免疫测定：在花生四烯酸的各种环氧合酶代谢产物中，前列腺素E2显示出多种生物活性，并且其产生被测定为跟踪环氧合酶表达。作为一种简便、灵敏的检测方法，我们尝试建立前列腺素E2的酶免疫分析法。我们开发了一种以乙酰胆碱酯酶为标记物的测定方法，其校准曲线的IC 50为4.5 pg/50 μl样品。

项目成果

期刊论文数量（30）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

N.Ueda, K.Tsuboi, S.Yamamoto: "Enzymological studies on endocannabinoid metabolism"International Symposium on Recent Biomedical Advances in Eicosanoid Research. Abstract. 33 (2002)

N.Ueda、K.Tsuboi、S.Yamamoto：“内源性大麻素代谢的酶学研究”二十烷酸研究最新生物医学进展国际研讨会。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

プロスタグランジンE_2の酵素免疫測定法

前列腺素E_2酶联免疫分析

DOI：
发表时间：
2004
期刊：
ビタミン 78(4)
影响因子：
0
作者：
Trembley;J.H.;Tatsumi;S.;Sakashita;E.;Loyer;P.;Slaughter;C.A.;Suzuki;H.;Endo;H.;Kidd;V.J.;Mayeda;A.;勝川路子
通讯作者：
勝川路子

R.Wiesner: "Suicidal inactivation of the rabbit 15-lipoxygenase by 15S-HPETE is paralleled by covalent modification of active site peptides"Free Radical Biology & Medicine. 34(2). 304-315 (2003)

R.Wiesner：“15S-HPETE 对兔子 15-脂氧合酶的自杀性失活与活性位点肽的共价修饰是平行的”自由基生物学