Quercetin coumaroyl glucorhamnoside from Ginkgo biloba

A new flavonoid glycoside was isolated from the leaves of Ginkgo biloba (Ginkgoaceae) and its structure elucidated as quercetin-3-O-α-(6‴-p-coumaroylglucosyl-β-1,4-rhamnoside).     

Camellianoside, a novel antioxidant glycoside from the leaves of Camellia japonica

A novel flavonol glycoside named camellianoside and three known flavonol glycosides were isolated from the leaves of Camellia japonica. The structure of camellianoside was established as quercetin-3-O-beta-D-xylopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->6)-O-beta-D-glucopyranoside by spectroscopic and chemical methods. The antioxidant activities of these glycosides evaluated by the diphenylpicrylhydrazyl (DPPH) radical scavenging reaction was higher than those of L-cysteine and L-ascorbic acid used as the reference antioxidants.     

Phytochemical investigation on Mercurialis annua

A new natural flavonol glycoside, isorhamnetin-3-rutinoside-4′-glucoside, together with rutin, narcissin, quercetin-3-(2^G-glucosyl)-rutinoside and isorhamnetin-3-rutinoside-7-glucoside, was identified from the MeOH extract of Mercurialis annua L. The structures were established on the basis of chemical and spectral (¹H and ¹³C NMR, FAB MS) data.     

Polyphenols from Cornulaca monacantha

The aerial parts of the Egyptian herb Cornulaca monacantha have yielded two new galloyltannin analogs (named monacanthin A and monacanthin B) characterized by protoctechuoyl moiety at C-6, and also a new flavonol glycoside, quercetin-4'-O-beta-D-galactoside. The structures of the new compounds were established by conventional analytical methods and confirmed by spectral analyses. Two known flavonoids, luteolin-7-O-rhamnoside, luteolin-7-O-glucoside, and two known gallotannins, penta-O-galloyl-beta-D-glucose and 1,2,3,6-tetra-O-galloyl-beta-D-glucose were also isolated.     

微生物糖基转移酶催化合成槲皮素糖苷及其抗炎活性评价

利用微生物来源的糖基转移酶GT-1、GT-2和BTGT-1对槲皮素进行糖基化修饰。研究发现这3种酶均能够催化以槲皮素和UDP-葡萄糖为底物生成槲皮素-3-O-β-D-葡萄糖（异槲皮苷）,GT-1、 GT-2和BTGT-1催化反应的产物生成率分别为5.33%、15.18%和63.82%。通过对槲皮素、异槲皮苷和槲皮素-N-乙酰-D-氨基葡萄糖进行水溶性以及体外细胞抗炎活性检测,发现槲皮素经糖基化后其水溶性得到较大提高,异槲皮苷和槲皮素-N-乙酰-D-氨基葡萄糖水溶性分别是槲皮素的13.8倍和15.4倍。同等浓度下槲皮素糖苷对RAW264.7 细胞的毒性作用低于槲皮素,且3种化合物在一定浓度范围内对LPS诱导RAW264.7细胞释放NO、 IL-1β、IL-6 都有显著的抑制作用,且呈剂量依赖性,研究表明3种化合物都具有一定的抗炎作用。     

Properties of quercetin conjugates: modulation of LDL oxidation and binding to human serum albumin

Quercetin is an important dietary flavonoid with in vitro antioxidant activity. However, it is found in human plasma as conjugates with glucuronic acid, sulfate or methyl groups, with no significant amounts of free quercetin present. The antioxidant properties of the conjugates found in vivo and their binding to serum albumin are unknown, but essential for understanding possible actions of quercetin in vivo. We, therefore, tested the most abundant human plasma quercetin conjugates, quercetin-3-glucuronide, quercetin-3'-sulfate and isorhamnetin-3-glucuronide, for their ability to inhibit Cu(II)-induced oxidation of human low density lipoprotein and to bind to human albumin, in comparison to free flavonoids and other quercetin conjugates. LDL oxidation lag time was increased by up to four times by low (<2 microM) concentrations of quercetin-3-glucuronide, but was unaffected by equivalent concentrations of quercetin-3'-sulfate and isorhamnetin-3-glucuronide. In general, the compounds under study prolonged the lag time of copper-induced LDL oxidation in the order: quercetin-7-glucuronide > quercetin > quercetin-3-glucuronide = quercetin-3-glucoside > catechin > quercetin-4'-glucuronide > isorhamnetin-3-glucuronide > quercetin-3'-sulfate. Thus the proposed products of small intestine metabolism (quercetin-7-glucuronide, quercetin-3-glucuronide) are more efficient antioxidants than subsequent liver metabolites (isorhamnetin-3-glucuronide, quercetin-3'-sulfate). Albumin-bound conjugates retained their property of protecting LDL from oxidation, although the order of efficacy was altered (quercetin-3'-sulfate > quercetin-7-glucuronide > quercetin-3-glucuronide > quercetin-4'-glucuronide = isorahmnetin-3-glucuronide). Kq values (concentration required to achieve 50% quenching) for albumin binding, as assessed by fluorescence quenching of Trp214, were as follows: quercetin-3'-sulfate (approximately 4 microM)= quercetin > or = quercetin-7-glucuronide > quercetin-3-glucuronide = quercetin-3-glucoside > isorhamnetin-3-glucuronide > quercetin-4'-glucuronide (approximately 20 microM). The data show that flavonoid intestinal and hepatic metabolism have profound effects on ability to inhibit LDL oxidation and a lesser but significant effect on binding to serum albumin.     

Anaerobic transformation of quercetin-3-glucoside by bacteria from the human intestinal tract

From human feces two phenotypically different types of bacteria were isolated on quercetin-3-glucoside as carbon and energy source. Isolates of one type were identified as strains of Enterococcus casseliflavus. They utilized the sugar moiety of the glycoside, but did not degrade the aglycon further. The sugar moiety (4 mM) was fermented to 5.5 ± 2.1 mM formate, 2.1 ± 0.7 mM acetate, 1.6 ± 0.3 mM l-lactate, and 1.3 ± 0.4 mM ethanol. The second type of isolate was identified as Eubacterium ramulus. This organism was capable of degrading the aromatic ring system. Growing cultures of Eubacterium ramulus converted 5 mM quercetin-3-glucoside to 1.7 ± 0.6 mM 3,4-dihydroxyphenylacetic acid, 7.6 ± 1.0 mM acetate, and 4.0 ± 0.4 mM butyrate. Molecular hydrogen, 3,4-dihydroxybenzaldehyde, and ethanol were detected in small amounts. Phloroglucinol was a transient intermediate in the breakdown of quercetin-3-glucoside. Eubacterium ramulus did not grow on the aglycon quercetin or the ring-fission intermediate phloroglucinol, but cleaved the flavonoid ring system when glucose was present as a cosubstrate. The most probable number of quercetin-3-glucoside-degrading bacteria determined in nine human fecal samples was 10⁷–10⁹/g dry mass. Isolates from these experiments were all identified as Eubacterium ramulus. Received: 9 July 1998 / Accepted: 10 November 1998     

Phenolics in the Strawberry Root

Roots of four strawberry cultivars contained phenolics previouslyisolated from the fruits and leaves; others were found whichapparently have not been reported. Catechin, gallic acid, andthree biflavans were the most prominent as spots on paper chromatograms.Kaempherol-7-glucoside, quercetin-7-glucoside, an unidentifiedflavanone glycoside, ellagic acid, an ellagic acid derivative,a galloyl ester, and two other hydrolysable tannins were detected.A leucoanthocyanin, cyanidin-3-monoglucoside, chlorogenie acid,a scopoletin glycoside, a derivative of arbutin, DOPA (3,4-dihydroxyphenylalanine),and three unidentified polyphenols comprised the other phenolicscommon to the four cultivars. A xanthone was found in only oneof four cultivars, Howard 17.     

A Drosophila model for the screening of bioavailable NADPH oxidase inhibitors and antioxidants

NADPH oxidase is the major source of non-mitochondrial cellular reactive oxygen species (ROS), and also is reported to be a major cause of various diseases including atherosclerosis and hypertension. In order to screen a new curative reagent that can suppress NADPH oxidase activity, we developed a Drosophila melanogaster fly that would overexpress human Dual oxidase 2 (hDuox2), a member of the NADPH oxidase family, as a screening model. These flies (GMR-GAL4/UAS-hDuox2) had a high generation of ROS in the posterior region of the eye discs along with an easily recognizable rough-eye phenotype, which is an ideal and convenient marker for further screening steps. Moreover, the hDuox2-induced rough-eye phenotype can be rescued by feeding with a culture medium containing mulberry leaves (MLs), which reportedly have an antimetabolic effect. Some commercially available antioxidants such as quercetin-3-O-D-glucoside or quercetin-3-O-glucose-6'-acetate, or the naringin contained in MLs and other herbs, also have shown a similar suppressing effect on the rough-eye phenotype. Our results suggest that flavonoid glycoside is absorbed from the intestine and functions in the body of D. melanogaster as it does in mammalian models such as rats. Thus, the GMR-GAL4/UAS-hDuox2 fly line is a promising model for the screening of novel drugs such as NADPH oxidase inhibitors and/or antioxidants.     

Metabolic conversion of dietary flavonoids alters their anti-inflammatory and antioxidant properties

The notion that dietary flavonoids exert beneficial health effects in humans is often based on in vitro studies using the glycoside or aglycone forms of these flavonoids. However, flavonoids are extensively metabolized in humans, resulting in the formation of glucuronide, methyl, and sulfate derivatives, which may have different properties than their parent compounds. The goal of this study was to investigate whether different chemical modifications of the same flavonoid molecule affect its biological and antioxidant activities. Hence, we studied the anti-inflammatory effects of several major human metabolites of quercetin and (-)-epigallocatechin-3-O-gallate (EGCG) by assessing their inhibitory effects on tumor necrosis factor α (TNFα)-induced protein expression of cellular adhesion molecules in human aortic endothelial cells (HAEC). HAEC were incubated with 1-30 μM quercetin, 3'- or 4'-O-methyl-quercetin, quercetin-3-O-glucuronide, and quercetin-3'-O-sulfate or 20-100 μM EGCG, 4'-O-methyl-EGCG, and 4',4'-di-O-methyl-EGCG, prior to coincubation with 100 U/ml of TNFα. 3'-O-Methyl-quercetin, 4'-O-methyl-quercetin, and their parent aglycone compound, quercetin, all effectively inhibited expression of intercellular adhesion molecule-1 (ICAM-1) with IC(50) values (concentration required for 50% inhibition) of 8.0, 5.0, and 4.4 μM, respectively; E-selectin expression was suppressed to a somewhat lesser but still significant degree by all three compounds, whereas vascular cell adhesion molecule-1 (VCAM-1) was not affected. In contrast, quercetin-3-O-glucuronide (20-100 μM), quercetin-3'-O-sulfate (10-30 μM), and phenolic acid metabolites of quercetin (20-100 μM) did not inhibit adhesion molecule expression. 4',4'-Di-O-methyl-EGCG selectively inhibited ICAM-1 expression with an IC(50) value of 94 μM, whereas EGCG (20-60 μM) and 4'-O-methyl-EGCG (20-100 μM) had no effect. The inhibitory effects of 3'-O-methyl-quercetin and 4',4'-di-O-methyl-EGCG on adhesion molecule expression were not related either to inhibition of NF-κB activation or to their antioxidant reducing capacity. Our data indicate that flavonoid metabolites have different biological and antioxidant properties than their parent compounds, and suggest that data from in vitro studies using nonmetabolites of flavonoids are of limited relevance in vivo.     

Properties of Quercetin Conjugates: Modulation of LDL Oxidation and Binding to Human Serum Albumin

Quercetin is an important dietary flavonoid with in vitro antioxidant activity. However, it is found in human plasma as conjugates with glucuronic acid, sulfate or methyl groups, with no significant amounts of free quercetin present. The antioxidant properties of the conjugates found in vivo and their binding to serum albumin are unknown, but essential for understanding possible actions of quercetin in vivo. We, therefore, tested the most abundant human plasma quercetin conjugates, quercetin-3-glucuronide, quercetin-3′-sulfate and isorhamnetin-3-glucuronide, for their ability to inhibit Cu(II)-induced oxidation of human low density lipoprotein and to bind to human albumin, in comparison to free flavonoids and other quercetin conjugates. LDL oxidation lag time was increased by up to four times by low (<2?μM) concentrations of quercetin-3-glucuronide, but was unaffected by equivalent concentrations of quercetin-3′-sulfate and isorhamnetin-3-glucuronide. In general, the compounds under study prolonged the lag time of copper-induced LDL oxidation in the order: quercetin-7-glucuronide>quercetin>quercetin-3-glucuronide=quercetin-3-glucoside>catechin>quercetin-4′-glucuronide>isorhamnetin-3-glucuronide>quercetin-3′-sulfate. Thus the proposed products of small intestine metabolism (quercetin-7-glucuronide, quercetin-3-glucuronide) are more efficient antioxidants than subsequent liver metabolites (isorhamnetin-3-glucuronide, quercetin-3′-sulfate). Albumin-bound conjugates retained their property of protecting LDL from oxidation, although the order of efficacy was altered (quercetin-3′-sulfate>quercetin-7-glucuronide>quercetin-3-glucuronide>quercetin-4′-glucuronide=isorahmnetin-3-glucuronide). K_q values (concentration required to achieve 50% quenching) for albumin binding, as assessed by fluorescence quenching of Trp214, were as follows: quercetin-3′-sulfate (～4?μM)=quercetin≥quercetin-7-glucuronide>quercetin-3-glucuronide=quercetin-3-glucoside>isorhamnetin-3-glucuronide>quercetin-4′-glucuronide (～20?μM). The data show that flavonoid intestinal and hepatic metabolism have profound effects on ability to inhibit LDL oxidation and a lesser but significant effect on binding to serum albumin.     

New chromanone and acylphloroglucinol glycosides from the bracts of hops

A chromanone glycoside, 5-β-d-glucopyranosyloxy-7-hydroxy-2-isopropylchromanone, and two acylphloroglucinol glycosides, 2-(3-methylbutyryl)phloroglucinol-4,6-di-C-β-d-glucopyranoside and 2-isobutyrylphloroglucinol-1,5-di-O-β-d-glucopyranoside, along with 15 known compounds were isolated from the bracts of hops (Humulus lupulus L.). Their structures were elucidated based on spectroscopic data, including TOFMS and 1D/2D NMR.     

Immunochemical detection of flavonoid glycosides: development, specificity, and application of novel monoclonal antibodies

Flavonoid-rich diets are expected to decrease the risk of cardiovascular diseases. The localization and target sites of flavonoids underlying the protective mechanism in vivo have not been fully investigated because the methods for detection of flavonoids have been limited to chemical analysis such as high-performance liquid chromatography. To further understand the actions of flavonoids in vivo, we developed a novel methodology that immunochemically evaluates flavonoids using specific antibodies. Quercetin-3-glucuronide (Q3GA), a major metabolite in human plasma, was coupled with keyhole limpet hemocyanin. Alternatively, the sugar moiety of quercetin-3-glucoside (Q3G) was succinylated and then coupled with a carrier protein. Using these two immunogens, we finally obtained two monoclonal antibodies, mAb14A2 and mAb11G6, from the immunogen using Q3GA and Q3G, respectively. Competitive enzyme-linked immunosorbent assay showed the unique difference in the specificity between the two similar antibodies: mAb14A2 recognized several quercetin-3-glycosides including Q3G and rutin but mAb11G6 was highly specific to the Q3G structure. The macrophage-derived foam cells in human atherosclerotic lesions were significantly stained with mAb14A2 but scarcely with mAb11G6. These results showed that the anti-flavonoid glycoside antibodies are useful tools for evaluating their localization in tissues and that the specificities strongly depend on the immunogen design for synthesizing the hapten-protein conjugates.     

棉花苞叶光呼吸和PSII热耗散对土壤水分的响应

在新疆气候生态条件下, 采用膜下滴灌植棉技术, 设置不同滴灌水分处理, 研究了不同滴灌量条件下棉花(Gossypium hirsutum)苞叶和叶片碳同化、光呼吸作用、光系统II (PSII)热耗散作用及其光破坏防御机制的差异, 以揭示滴灌节水条件下棉花苞叶缓解光抑制的机理及与棉花抗旱特性的关系。结果表明: 棉花开花后苞叶及叶片在高温强光下实际光化学效率(Φ_PSII)显著降低, 发生明显的光抑制现象, 但苞叶的光抑制程度较叶片轻; 与正常滴灌量处理相比, 节水滴灌条件下棉花水分亏缺, 叶片净光合速率(P_n)、Φ_PSII、光呼吸(P_r)、光化学猝灭系数(q_P)降低, 非光化学猝灭系数(NPQ)升高, 叶片光抑制程度加重, 而苞叶P_n、Φ_PSII、P_r、q_P、NPQ变化不大, 与正常滴灌量处理相比, 光抑制程度无显著差异。苞叶光呼吸速率与光合速率的比值(P_r/P_n)显著高于叶片; 滴灌节水条件下棉花适度水分亏缺对苞叶光呼吸及P_r/P_n无显著影响。高温强光下, 棉花节水滴灌对叶片PSII量子产量的转化与分配影响显著, 但对苞叶的影响不显著; 苞叶非调节性能量耗散的量子产量(Y(NPQ))高于叶片, 因此能有效地将PSII的过剩光能以热的形式耗散。综上所述, 与叶片相比, 苞叶对轻度水分亏缺不敏感, 是棉花适应干旱逆境较强的器官, 苞叶光呼吸和热耗散作用对光破坏防御具有重要意义。     

Protective effect of quercetin against cigarette tar extract-induced impairment of erythrocyte deformability

Quercetin (3,3',4',5,7-pentahydroxyflavone) is one of the most abundant flavonol-type flavonoids rich in diet and suggested to possess a beneficial role in blood circulation. This study was conducted to know the effect of quercetin aglycone and one of its possible metabolite, quercetin-3-O-beta-D-glucuronide on cigarette tar extract-induced impairment of erythrocyte deformability. Erythrocyte suspension containing quercetin aglycone, quercetin-3-O-beta-D-glucuronide or quercetin-3-O-beta-D-glucoside was forced to flow through microchannels with equivalent diameter of 7 &mgr;m and its transit time was measured as an index of erythrocyte deformability using microchannel array method. Both quercetin aglycone and quercetin-3-O-beta-D-glucuronide, but not quercetin-3-O-beta-D-glucoside, substantially increased erythrocyte deformability indicating that the former two compounds affect the physicochemical state of erythrocyte by interacting with its membranes. Aqueous cigarette tar extract caused marked decrease in erythrocyte deformability with concomitant increase of membranous lipid peroxidation. In that case, quercetin aglycone suppressed the impairment of erythrocyte deformability as well as membranous lipid peroxidation. The same effect was found in quercetin-3-O-beta-D-glucuronide, eventhough its effect was lower than that of quercetin aglycone. Thus, not only quercetin aglycone but also its conjugate metabolite protects erythrocyte membrane from the damage of smoking by scavenging reactive oxygen species generated from cigarette tar. Intake of quercetin-rich food may be helpful to protect membranous damage in erythrocytes from smoking.     

Production of a novel quercetin glycoside through metabolic engineering of Escherichia coli

Most flavonoids exist as sugar conjugates. Naturally occurring flavonoid sugar conjugates include glucose, galactose, glucuronide, rhamnose, xylose, and arabinose. These flavonoid glycosides have diverse physiological activities, depending on the type of sugar attached. To synthesize an unnatural flavonoid glycoside, Actinobacillus actinomycetemcomitans gene tll (encoding dTDP-6-deoxy-L-lyxo-4-hexulose reductase, which converts the endogenous nucleotide sugar dTDP-4-dehydro-6-deoxy-L-mannose to dTDP-6-deoxytalose) was introduced into Escherichia coli. In addition, nucleotide-sugar dependent glycosyltransferases (UGTs) were screened to find a UGT that could use dTDP-6-deoxytalose. Supplementation of this engineered strain of E. coli with quercetin resulted in the production of quercetin-3-O-(6-deoxytalose). To increase the production of quercetin 3-O-(6-deoxytalose) by increasing the supplement of dTDP-6-deoxytalose in E. coli, we engineered nucleotide biosynthetic genes of E. coli, such as galU (UTP-glucose 1-phosphate uridyltransferase), rffA (dTDP-4-oxo-6-deoxy-d-glucose transaminase), and/or rfbD (dTDP-4-dehydrorahmnose reductase). The engineered E. coli strain produced approximately 98 mg of quercetin 3-O-(6-deoxytalose)/liter, which is 7-fold more than that produced by the wild-type strain, and the by-products, quercetin 3-O-glucose and quercetin 3-O-rhamnose, were also significantly reduced.     

Flavonoids in translucent bracts of the Himalayan<Emphasis Type="Italic"> Rheum nobile</Emphasis> (Polygonaceae) as ultraviolet shields

UV-absorbing substances were isolated from the translucent bracts of Rheum nobile, which grows in the alpine zone of the eastern Himalayas. Nine kinds of the UV-absorbing substances were found by high performance liquid chromatography (HPLC) and paper chromatography (PC) surveys. All of the five major compounds are flavonoids, and were identified as quercetin 3-O-glucoside, quercetin 3-O-galactoside, quercetin 3-O-rutinoside, quercetin 3-O-arabinoside and quercetin 3-O-[6-(3-hydroxy-3-methylglutaroyl)-glucoside] by UV, ¹H and ¹³C NMR, mass spectra, and acid hydrolysis of the original glycosides, and direct PC and HPLC comparisons with authentic specimens. The four minor compounds were characterised as quercetin itself, quercetin 7-O-glycoside, kaempferol glycoside and feruloyl ester. Of those compounds, quercetin 3-O-[6-(3-hydroxy-3-methylglutaroyl)-glucoside] was found in nature for the first time. The translucent bracts of R. nobile accumulate a substantial quantity of flavonoids (3.3–5 mg per g dry material for the major compounds). Moreover, it was clarified by quantitative HPLC survey that much more of the UV-absorbing substances is present in the bracts than in rosulate leaves. Although the flavonoid compounds have been presumed to be the important UV shields in higher plants, there has been little characterisation of these compounds. In this paper, the UV-absorbing substances of the Himalayan R. nobile were characterised as flavonol glycosides based on quercetin.     

Taxodione, a DNA-binding compound from Taxodium distichum L. (Rich.)

8-beta-Hydroxypimar-15-en-19-oic acid (1), taxodione (2), 6,7-dehydro-8-hydrotaxodone (3), quercetin-3-O-beta-D-glucopyranoside (4), and shikimic acid (5) were isolated from the leaves of Taxodium distichum L. (Rich.) for the first time. Previously reported compounds [beta-sitosterol (6), isorhamnetin (7), quercetin (8), isorhamnetin-3-O-alpha-arabinofuranoside (9), quercetin-3-O-a-arabinofuranoside (10)] have also been isolated. The activity of taxodione as an inhibitor for hepatic stellate cells was determined. The antitumour activity of 2, 3, and 5 using a DNA affinity probe was examined.     

Photosynthesis of bract and its contribution to seed maturity inCarpinus laxiflora

Light saturated net photosynthesis was measured in bracts and leaves ofCarpinus laxiflora, the major species in secondary forests in cool and intermediate temperate zones in Japan. The maximum net photosynthesis of leaves and bracts was essentially constant from May to early August and decreased gradually thereafter. For bracts, it was 3.2 μmol m⁻²s⁻¹, approximately half that for the leaves. The photosynthesis of bracts would thus appear to contribute significantly to seed maturity. The estimated production of bract based on the photosynthesis would make seeds (3 mg dry weight) mature for 37 days, assuming all photosynthate of the bracts to have been distributed in the seeds only. This was quite consistent with the growth curve for the seeds. A mast year phenomenon is discussed in relation to bract photosynthesis and leaf number.     

Acylated flavonoid glycosides from Bassia muricata.

From the arial parts of Bassia muricata, two acylated flavonoid glycosides quercetin-3-O-(6"-caffeoyl)-sophoroside and quercetin-3-O-(6"-feruloyl)-sophoroside have been isolated together with two known flavonoid glycosides quercetin-3-O-sophoroside and quercetin-3,7-O-beta-diglucopyranoside, as well as four known triterpenoidal saponins, oleanolic acid-3-O-beta-glucopyranoside, chikusetsusaponin IVa, chikusetsusaponin IVa methyl ester and oleanolic acid-3,28-beta-diglucopyranoside. The structures of the isolated compounds were verified by means of MS and NMR spectral analyses.