Hydrangeae Dulcis Folium Research Articles

Comparison of Growth in Hydrangea macrophylla var. thunbergii Grown in Different Soil pH and Quantitative Analysis of Its Sweetness-Related Constituents.

Dried and fermented (processed) leaves of Hydrangea macrophylla Seringe var. thunbergii Makino (Hydrangeae Dulcis Folium) are currently used as a crude drug with a sweet taste for diabetic patients and as an oral refrigerant. The sweet taste of this crude drug is primarily attributed to phyllodulcin. However, there are currently no standards for the cultivation of H. macrophylla var. thunbergii and the isolation and production of the primary constituents of this crude drug. In the present study, we prepared five types of soils with different pH values (pH 7.5-5.0) and investigated the effects of these soils on the growth of this plant. The contents of phyllodulcin and its glycoside, phyllodulcin 8-O-β-D-glucopyranoside, in the leaves of plants grown in these soils were quantified. Furthermore, the correlation between the sweetness of Hydrangeae Dulcis Folium and phyllodulcin was investigated. The results showed that soils with pH ranging from 7.0 to 5.5 was not only suitable for plant growth but also increased the content of phyllodulcin and phyllodulcin 8-O-β-D-glucopyranoside in the leaves. Altogether, these findings could be useful for the development of high-quality Hydrangeae Dulcis Folium.

Hydrangeae Dulcis Folium Attenuates Physical Stress by Supressing ACTH-Induced Cortisol in Zebrafish.

To determine the effects of Hydrangeae Dulcis Folium (EHDF) on physical stress, changes in the whole-body cortisol level and behaviour in zebrafish (Danio rerio). One hundred and seventy-four fish were randomly divided into 4 [adrenocorticotropin hormone (ACTH) challenge test: 4 fish per group] or 6 groups (behavioural test: 10-12 fish per group, whole-body cortisol: 4 fish per group). Net handling stress (NHS) was used to induce physical stress. Fish were treated with vehicle or EHDF (5-20 mg/L) for 6 min before they were exposed to stress. And then, fish were sacrificed for collecting body fluid from whole-body or conducted behavioural tests, including novel tank test and open field test, and were evaluated to observe anxiety-like behaviours and locomotion. In addition, to elucidate the mode of action of the anti-stress effects of EHDF, ACTH (0.2 IU/g, i.p.) challenge test was performed. The increased anxiety-like behaviours in novel tank test and open field test under stress were prevented by treatment with EHDF at 5-20 mg/L (P <0.05). Moreover, compared with the unstressed group, which was not treated with NHS, the whole-body cortisol level was significantly increased by treatment with NHS (P <0.05). Compared with the NHS-treated stressed control group, pre-treatment with EHDF at concentrations of 5-20 mg/L for 6 min significantly prevented the NHS-increased whole-body cortisol level (<0.05). In addition, ACTH challenge test showed that EHDF completely blocked the effects of ACTH on cortisol secretion (P <0.05). EHDF may be a good antistress candidate and its mechanism of action may be related to its positive effects on cortisol release.

수국 추출물이 알코올로 유도한 기억 장애 및 long-term potentiation 억제에 미치는 영향

다량의 에탄올을 섭취하면 기억 상실로 이어질 수 있으며, 종종 blackout으로 나타난다. Blackout의 불균형은 알코올 소비에 있어 다양한 사회 문제의 주요 원인이 될 수 있다. 그러나 이러한 알코올 유발 문제를 예방하는 치료법은 아직 존재하지 않는다. Hydrangeae dulcis folium은 Hydrangea serrata Seringe의 잎을 발효가공을 통해 만든 민간약 또는 차이다. 본 연구에서는 에탄올로 유도한 정신적 결핍에 대한 Hydrangeae dulcis folium의 에탄올 추출물(EHDF)의 효과를 평가하였다. 행동적 결핍 또는 장애를 테스트하기 위해 마우스에서 물체 인식 테스트가 수행하였다. 또한 시냅스 결손을 평가하기 위해, 마우스 해마 조각에서 에탄올에 취약한 것으로 알려져 있고 에탄올로 유발한 기억 상실과 관련이 있는 N-methyl-D-aspartate (NMDA) 수용체-매개 흥분성 시냅스 후 전위 및 long-term potentiation (LTP)을 측정하였다. 본 연구에서 에탄올(1 g/kg, i.p.)은 물체 인식 메모리를 손상 시켰지만, EHDF (10 또는 30 mg/kg)는 물체 인식 테스트에서 이러한 장애를 극복하였다. 흥미롭게도, EHDF (<TEX>$30{\mu}g/ml$</TEX>)는 해마 절편에서 에탄올 처리 후 억제되었던 LTP 및 NMDA 수용체 매개 시냅스 전달을 유의하게 개선시켰다. EHDF는 에탄올에 의해 유발된 물체 인식 기억력 결핍을 개선하였고, 또한 EHDF는 해마 절편에서 에탄올 유도성 LTP 및 NMDA 수용체 매개성 시냅스 전달을 상당히 개선시켰다. Consumption of high doses of ethanol can lead to amnesia, which often manifests as a blackout. This incoordination of blackout may be a major cause in various social problems in alcohol consumption. However, there is still no treatment for preventing these alcohol-induced problems. Hydrangeae dulcis folium is a drug or a tea which is made from the fermented and dried leaves of Hydrangea serrata Seringe. The present study, we tested the ethanol extract of the Hydrangeae dulcis folium (EHDF) on ethanol-induced psychological deficits. To test behavioral deficits, an object recognition test was conducted using a mouse model. To evaluate synaptic deficits, N-methyl-D-aspartate (NMDA) receptor-mediated excitatory postsynaptic potential EPSP and long-term potentiation (LTP) in the mouse hippocampal slices were tested, as they are known to be vulnerable to ethanol and are associated with ethanol-induced amnesia. In the tests, ethanol (1 g/kg, i.p.) impaired object recognition memory, but EHDF (10 or 30 mg/kg) prevented this impairment in object recognition test. Interestingly, EHDF (<TEX>$30{\mu}g/ml$</TEX>) significantly ameliorated ethanol-induced LTP and NMDA receptor-mediated synaptic transmission in the hippocampal slices. EHDF prevented ethanol-induced object recognition memory deficits induced by ethanol. Interestingly, EHDF significantly ameliorated ethanol-induced LTP and NMDA receptor- mediated synaptic transmission in the hippocampal slices.

ChemInform Abstract: Search for Antidiabetic Constituents of Medicinal Food

AbstractReview: 32 refs.

Search for Antidiabetic Constituents of Medicinal Food

Many foods are known to have not only nutritive and taste values but also medicinal effects. In Chinese traditional medicine, the treatment using medicinal foods has been recommended highly. Recently, we examined the effects of the extract and constituents of several medicinal foods on experimental models of diabetes. In this paper, we focus on the bioactive constituents of four medicinal foods, namely the antidiabetic constituents from 1) the roots, stems and leaves of Salacia plants, 2) the male flowers of Borassus flabellifer, 3) the flower buds of Camellia sinensis, 4) the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium).

Hydrangeic acid from the processed leaves of Hydrangea macrophylla var. thunbergii as a new type of anti-diabetic compound

Hydrangeic acid (3–100 μM), a stilbene constituent of the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium), promoted adipogenesis of 3T3-L1 cells. Hydrangeic acid significantly increased the amount of adiponectin released into the medium, the uptake of 2-deoxyglucose into the cells, and the translocation of glucose transporter 4 (GLUT4). Hydrangeic acid also increased mRNA levels of adiponectin, peroxisome proliferator-activated receptor γ2 (PPARγ2), GLUT4, and fatty acid-binding protein (aP2) while it decreased the expression of tumor necrosis factor α (TNF-α) mRNA. However, it did not activate PPARγ directly different from troglitazone in a nuclear receptor cofactor assay system. Furthermore, hydrangeic acid significantly lowered blood glucose, triglyceride, and free fatty acid levels after its administration for 2 weeks at a dose of 200 mg/kg/day (p.o.) in KK-A y mice.

Inhibitory effects of thunberginols A and B isolated from Hydrangeae Dulcis Folium on mRNA expression of cytokines and on activation of activator protein-1 in RBL-2H3 cells

Previously, thunberginols A and B from the processed leaves of Hydrangeae macrophylla var. thunbergii (Hydrangea dulcis folium) substantially inhibited the degranulation caused by antigen and calcium ionophore A23187, and the release of tumor necrosis factor (TNF)-alpha and interleukin (IL)-4 by antigen in RBL-2H3 cells. In the present study, we examined the effect of thunberginol B on the expression of mRNA of several cytokines [ILs-2, 3, 4 and 13, TNF-alpha and granulocyte/macrophage-colony stimulating factor (GM-CSF)] and effects of thunberginols A and B on activator protein (AP)-1 composed of c-jun and c-fos, which is essential for the expression of the cytokine mRNA, in RBL-2H3 cells. Thunberginol B inhibited up-regulated genes of all cytokines, and thunberginols A and B (30 microM) inhibited the phosphorylation of c-jun and expression of c-fos mRNA and phosphorylation of extracellular signal-regulated kinases (ERK1/2). In addition, the profile of gene expression by thunberginol B was similar to that by luteolin, a natural flavone with a potent anti-allergic effect.

New type of anti-diabetic compounds from the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium)

Two 3-phenyldihydroisocoumarins (hydrangenol and phyllodulcin), a 3-phenylisocoumarin (thunberginol A), and a stilbene (hydrangeaic acid) from the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium) promoted adipogenesis of 3T3-L1 cells. Hydrangenol, a principal constituent, significantly increased the amount of adiponectin released into the medium and mRNA levels of adiponectin, peroxisome proliferator-activated receptor γ2 (PPARγ2), and glucose transporter 4 (GLUT4), while it decreased the expression of interleukin 6 (IL-6) mRNA. Furthermore, hydrangenol significantly lowered blood glucose and free fatty acid levels 2 weeks after its administration at a dose of 200 mg/kg/d in KK-A y mice.

Inhibitory Effects of Thunberginols A, B, and F on Degranulations and Releases of TNF-.ALPHA. and IL-4 in RBL-2H3 Cells

Thunberginols A, B, and F from the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium) substantially inhibited the degranulations by antigen and calcium ionophore A23187, and the releases of TNF-alpha and IL-4 by antigen in RBL-2H3 cells. Phyllodulcin and hydrangenol also showed significant inhibition for the antigen-induced degranulations, but their effects were weaker than those of thunberginols A, B, and F. Among them, thunberginol B showed the most potent activity. With regard to structural requirements of thunberginols for the activity, the 3,4-double bond was essential for the strong activity and the 6-hydroxyl group and lactone ring enhanced the activity. Thunberginols A, B, and F inhibited increase in intracellular free Ca2+ levels, which is an essential process for the degranulation and production of cytokines, in RBL-2H3 cells induced by antigen, but not by calcium ionophore A23187. These results suggested that these active compounds inhibited the degranulation processes both before and after increase in intracellular free Ca2+ levels.

Suppression by Hydrangeae Dulcis Folium of D-galactosamine-induced liver injury in vitro and in vivo.

Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla SER. var. thunbergii MAKINO, suppressed D-galactosamine-induced liver injury by 85.2% when added to the diet at 1% and fed to rats for fifteen days. The hepatoprotective effect is more potent than that of a milk thistle extract and turmeric powder. Some fractionated extracts showed hepatoprotective activity in the D-galactosamine-induced in vitro liver injury model.

New anti-malarial flavonol glycoside from hydrangeae dulcis folium

Bioassay-guided fractionation of the MeOH extract of Hydrangeae Dulcis Folium resulted in isolation of a new flavonol glycoside and two known congeners as anti-malarial principles. These flavonol glycosides showed characteristic proliferation inhibition of Plasmodium falciparum at significantly low concentration without showing any cytotoxicity. In addition, several naturally occurring flavonol glycosides were also shown to exert similar anti-malarial behavior.

Structure-Requirements of isocoumarins, phthalides, and stilbenes from hydrangeae dulcis folium for inhibitory activity on histamine release from rat peritoneal mast cells

We examined the structure–activity relationships of isocoumarins, phthalides and stilbenes isolated from Hydrangeae Dulcis Folium and related compounds for the inhibition of histamine release in rat peritoneal mast cells. The activities of isocoumarins such as thunberginols A and B were more potent than those of dihydroisocoumarins such as hydrangenol and thunberginol G. The double bond at the 3-position seemed to be essential to potentiate the activity. The hydroxyl groups at the 8-, 3′- and 4′-positions of isocoumarin were essential for the activity, while the hydroxyl group at the 6-position was scarcely needed. Since the activities of benzylidenephthalides such as thunberginol F were more potent than those of hydramacrophyllols A and B, the presence of a double bond at the 3-position was needed to increase the activity. Moreover, the hydroxyl group at the 8-position was essential for the activity. On the time course study, thunberginols A, B and F completely inhibited histamine release by pretreatment at 100μM for 1 to 15min, whereas DSCG inhibited histamine release only following 1-min pretreatment at 1000μM. These results suggested that the mechanisms of the inhibitory effect of thunberginols are different from that of DSCG. ©

Effects of phyllodulcin, hydrangenol, and their 8-O-glucosides, and thunberginols A and F from Hydrangea macrophylla SERINGE var. thunbergii MAKINO on passive cutaneous anaphylaxis reaction in rats.

We examined the antiallergic effects of phyllodulcin, hydrangenol, and their 8-O-glucosides, and thunberginols A and F isolated from the processed leaves (Hydrangeae Dulcis Folium) and dried leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO using the passive cutaneous anaphylaxis (PCA) reaction. With the exception of phyllodulcin, these constituents were found to significantly inhibit the PCA reaction. Although thunberginol A showed the most potent inhibitory effect, hydrangenol was considered to be the principal antiallergic component in the processed leaves, after taking into account their contents.

Effect of thunberginol A from Hydrangeae Dulcis Folium on signal transduction pathway for mast cell degranulation

Effect of thunberginol A from Hydrangeae Dulcis Folium on signal transduction pathway for mast cell degranulation

The Role of Thunberginol A, an Isocoumarin Constituent of Hydrangeae Dulcis Folium, on the Signal Transmission Pathway for Rat Mast Cell Degranulation.

The role of the signal transmission pathway of thunberginol A (TA) in mast cell degranulation was examined using rat peritoneal mast cells. First of all, we investigated the cellular distribution of TA using fluorescent microscopy. This indicated that TA is immediately incorporated into cells and distributes in cytosol around the nucleus. We then investigated the effect of TA on mast cell protein tyrosine phosphorylation, which is part of the signal transduction cascade for degranulation. TA non-specifically inhibited the tyrosine phosphorylation induced by compound 48/80 (Co. 48/80), at 10 to 100 microM, and orthovanadate/hydrogen peroxide at more than 50 microM in a dose-dependent manner. As far as the intracellular Ca2+ change in fluo-3-loaded cells was concerned, TA (10 microM) suppressed the rise in Ca2+ induced by antigens, ionomycin and thapsigargin, while TA did not suppress the rise induced by Co. 48/80. This evidence suggests that TA mainly inhibits extracellular Ca2+ influx, but TA does not act on the intracellular Ca2+ mobilization from the endoplasmic reticulum. We also investigated the influence of TA on the cytoskeleton and membrane changes using mast cells and erythrocytes. TA (10 microM) inhibited the cytoskeletal assembly formation in dicyanovinyl julolidin-loaded mast cells induced by Co. 48/80. Moreover, TA suppressed the hypotonic hemolysis of erythrocytes, from 3 to 1000 microM, in a dose-dependent manner. These results suggest that inhibition of protein tyrosine phosphorylation, extracellular Ca2+ influx and cytoskeletal assembly formation, and membrane stabilization are involved in the inhibitory effect of TA in mast cell degranulation.

Immunomodulatory activity of thunberginol A and related compounds isolated from Hydrangeae Dulcis Folium on splenocyte proliferation activated by mitogens.

We investigated the immunomodulatory effects of antiallergic constituents from Hydrangeae Dulcis Folium, the processed leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO, on splenocyte proliferation in mice. Thunberginol A and hydrangenol significantly suppressed T lymphocyte proliferation induced by concanavalin A. Thunberginol A also suppressed B lymphocyte proliferation induced by lipopolysaccharide, but other constituents induced significant increases. These inhibitory effects of thunberginol A on splenocyte proliferation seemed to contribute to the suppressive effect on type IV allergy.

Development of bioactive functions in Hydrangeae Dulcis Folium. VII. Immunomodulatory activities of thunberginol A and related compounds on lymphocyte proliferation.

Immunomodulatory activities of constituents isolated from Hydrangeae Dulcis Folium and their related compounds on lymphocyte proliferation were investigated using splenocytes and lymph node cells. Thunberginol A (TA) significantly suppressed B lymphocyte proliferation stimulated by lipopolysaccharide (LPS) at 10(-5) M. However TA at a lower concentration (10(-6) M) and the other compounds, except hydrangenol and 3'-hydroxyhydrangeaic acid, tended to potentiate B lymphocytes proliferation (10(-5) M). On the other hand, thunberginol A significantly suppressed T lymphocyte proliferation stimulated by concanavalin A (Con A), but did not suppress phytohemagglutinin (PHA). Hydrocortisone and cyclosporin A strongly suppressed T lymphocyte proliferation induced by both Con A and PHA. These results suggest that thunberginol A acts on both B and T lymphocytes and may have a suppressive mechanism different from the known immunosuppressants. The cytotoxicity of TA for splenocytes seemed weaker than known immunosuppressants resulting from viability tests using MTT assay and the measurement of lactate dehydrogenase (LDH) released in the medium. Moreover, TA suppressed antigen-specific T lymphocyte proliferation in mice lymph node cells immunized with keyhole limpet hemocyanin (KLH). Thus, these findings show that TA has a suppressive effect on lymphocyte activation, and this inhibitory effect of TA seems to contribute to its suppressive effect on type IV allergies.

ChemInform Abstract: Development of Bioactive Functions in Hydrangeae Dulcis Folium. Part 6. Syntheses of Thunberginols A and F and Their 3′‐Deoxy‐Derivatives Using Regiospecific Lactonization of Stilbene Carboxylic Acid: Structures and Inhibitory Activity on Histamine Release of Hydramacrophyllols A and B.

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Development of bioactive functions in Hydrangeae dulcis folium. VI. Syntheses of thunberginols A and F and their 3'-deoxy-derivatives using regiospecific lactonization of stilbene carboxylic acid: structures and inhibitory activity on histamine release of hydramacrophyllols A and B.

Lactonization reaction of 2-carboxystilbene mediated by copper(II) chloride proceeded regiospecifically to give the five-membered lactone, while the bromolactonizations using N-bromosuccinimide and anodic oxidation were found to furnish the six-membered lactone. Using these regiospecific lactonization reactions as a key step, antiallergic and antimicrobial isocoumarins and the benzylidenephthalides thunberginols A and F and their 3'-deoxyanalogs were synthesized from phyllodulcin and hydrangenol. Two phthalides called hydramacrophyllols A and B were isolated from Hydrangeae Dulcis Folium and their stereostructures were determined on the basis of physicochemical and chemical evidence, which included the syntheses of hydramacrophyllols A and B from hydrangenol by the application of the lactonization method using copper(II) chloride. In addition, hydramacrophyllols A and B were found to exhibit an inhibitory effect on the histamine release from rat peritoneal exudate cells induced by antigen-antibody reaction.

Development of bioactive functions in hydrangeae dulcis folium. V. On the antiallergic and antimicrobial principles of hydrangeae dulcis folium. (2). Thunberginols C, D, and E, thunberginol G 3'-O-glucoside, (-)-hydrangenol 4'-o-glucoside, and (+)-hydrangenol 4'-O-glucoside.

Following the characterization of thunberginols A, B, and F, six bioactive principles, thunberginols C, D, and E, thunberginol G 3'-O-glucoside, (-)-hydrangenol 4'-O-glucoside, and (+)-hydrangenol 4'-O-glucoside, were isolated from Hydrangeae Dulcis Folium, the processed leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO, together with four kaempferol and quercetin oligoglycosides. Their chemical structures have been determined on the basis of chemical and physicochemical evidence. Thunberginols C, D, E, and G and (-)-hydrangenol 4'-O-glucoside showed antiallergic activity in the in vitro bioassay using the Schultz-Dale reaction. These components also exhibited inhibitory activities on the histamine release from rat mast cells and on the histamine-induced contraction in isolated guinea pig tracheal chain. In addition, thunberginols C, D, E, and G showed antimicrobial activities against oral bacteria.