Effect of polyphenols from Syringa vulgaris on blood stasis syndrome.

In this study, we employed a previously developed in vivo assay system to determine whether the flowers and leaves of Syringa vulgaris (S. vulgaris; commonly known as "lilac") can prevent blood stasis syndrome, known as oketsu in Japanese. This syndrome is considered an important pathology in traditional Chinese and Japanese medicine, and is related to diseases such as peripheral vascular disorders, blood vessel inflammation, and platelet aggregation, whose severities are augmented owing to lipid peroxidation, free radicals, and oxidative stress. The assay system employed in this study monitored the blood flow decrease in the tail vein of mice subjected to sensitization with hen egg white lysozyme. Through bioassay-guided fractionation of different S. vulgaris extracts, five polyphenols were isolated and identified. Among them, quercetine 3-glucoside, quercetin 3-rutinoside, and acteoside were identified as active compounds, as they significantly mitigated blood flow reduction. These findings indicate that the polyphenols obtained from S. vulgaris could be useful for preventing oketsu and improve the quality of life of individuals with disorders and diseases such as gynecopathy, cold sensitivity, poor circulation, allergy, and lifestyle-related diseases.

Allergy-preventive effects of linarinic acid and its tetrahydropyrrolo[2,1-b]quinazoline derivatives isolated from Linaria vulgaris.

Linarinic acid, (-)-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazoline-1-carboxylic acid (4a), was isolated from the ethanol extract of Linaria vulgaris Mill. In our previous study, a series of tetrahydropyrrolo[2,1-b]quinazoline derivatives 4b, 4c, 5a, 5b, 6a and 6b that were structurally related to 4a and evaluated as neuroprotective agents were synthesized. The aim of the present study was to investigate the novel features of these compounds. We examined their allergy-preventive effects using an in vivo assay system we developed previously, that monitors a decrease in blood flow in the tail vein of mice subjected to sensitization with hen egg-white lysozyme. We observed that 4a and its three derivatives, amide (6a), ester (5a), bromine (4b), and alcohol substituent (6b), showed significant allergy-preventive activities. The study confirmed the allergy-preventive activity of tetrahydropyrrolo[2,1-b]quinazoline derivatives by comprehensively monitoring the specific blood flow decrease occurring in the induction phase of allergy. This finding may aid in the development of new agents for the treatment of allergic diseases such as atopic dermatitis, allergic asthma, and hay fever.

Effects of pharmacological inhibition of plasminogen binding on liver regeneration in rats.

The fibrinolysis system is thought to play an important role in liver regeneration. We previously found that plasminogen (Plg) is localized to the cell surface of regenerating liver tissue as well as proliferating hepatocytes in vitro. Here, we investigated the significance of Plg binding to the cell surface during liver regeneration. Pre-administration of tranexamic acid (TXA), which is a competitive inhibitor of Plg binding, to hepatectomized rats mildly delayed restoration of liver weight in vivo. Although binding of Plg to the cell membrane decreased following TXA administration, TXA showed little effect on hepatocyte proliferation in rats. We also discovered that Plg treatment did not stimulate proliferation of primary rat hepatocytes in vitro. These results suggest that Plg/plasmin potentiates liver regeneration via a pathway distinct from those through which hepatocyte proliferation is stimulated.

Development of an Assay Method to Search for Compounds Inhibiting Stress-Enhanced Allergy.

Stress exacerbates allergic disorders such as atopic dermatitis and asthma. It is also an important factor affecting blood flow (BF). Allergic reactions also affect blood flow. For example, we observed that mice sensitized with hen egg-white lysozyme (HEL) have decreased BF during the allergy induction phase. Based on this finding, we established a model for evaluating chronic restraint stress-enhanced allergies. Mice were sensitized with 12.5 µg/head of HEL on day 0, then restrained for 90 min daily on days 1-3, 5, and 6 in a modified 50 mL polystyrene conical centrifuge tube with multiple air holes for ventilation. We used the decrease in BF during that time as a guide for developing an in vivo assay for substances that can inhibit stress-enhanced allergies. Finally, we demonstrated the utility of the new method by testing crude drugs that are used solely or in combination with other crude drugs to treat stress-related illness and neuropsychiatric symptoms. Our model should be useful for identifying potential anti-stress-enhanced allergy drugs.

Allergy-preventive effects of chlorogenic acid and iridoid derivatives from flower buds of Lonicera japonica.

Allergy-preventive activity of flower buds of Lonicera japonica THUNB. was found in the 35% EtOH extract (LJ) using an in vivo assay, The assay system uses monitoring of a decrease in blood flow (BF) in the tail vein of mice subjected to sensitization with hen-egg white lysozyme (HEL). Bioassay-guided fractionation of the 35% EtOH extract led to isolation of chlorogenic acid (1) and three known iridoid derivatives, loganin (2), secoxyloganin (3) and sweroside (4), all of which inhibited the BF decrease. This suggested that the flower buds of L. japonica and compounds isolated from them have allergy-preventive properties. The structure-activity relationship of iridoid derivatives, morroniside (5), geniposide (6), asperuloside (7), aucubin (8) and catalpol (9), were also tested using the same bioassay method. Compounds 2-5 and 9 having the sp(3) atom at C-8 showed an allergy-preventive effect, while compounds 6, 7 and 8 having a double bond at C-7, C-8 did not.

Preventive effects of the extract of kinka-cha, a folk tea, on a rat model of metabolic syndrome.

Kinka-cha (dried leaf of Camellia nitidissima) is used as a folk tea for detoxication, diuresis and antihypertension. In the present study, we evaluated the extract of kinka-cha on metabolic, vascular and oxidative stress parameters in a model of metabolic syndrome, SHR/NDmcr-cp/cp (SHR/cp) rats that manifest hypertension, obesity, glucose intolerance and hyperlipidemia. Treatment with the extract of kinka-cha alleviated the increase in blood pressure, decrease in tail blood flow and elevated serum oxidative stress marker levels including lipid peroxides, 8-hydroxy-2'-deoxyguanosine, 3-nitrotyrosine and 3-chlorotyrosine. However, kinka-cha did not affect weight gain, hyperglycemia and hyperlipidemia, nor the relaxation responses of the aorta mesenteric artery, thoracic aortas and tail vein, and blood clotting and platelet aggregation. These results suggest that kinka-cha can help reduce the risk of developing metabolic syndrome, possibly due to the presence of antioxidants.

Allergy-preventive effects of the flowers of Impatiens textori.

The allergy-preventive activity of a 35% EtOH extract (IT) of flowers of Impatiens textori MIQ. was demonstrated in a continuing search for allergy-preventive substances from natural sources. The evaluation of its activity used an in vivo assay method for monitoring the blood flow decrease in the tail vein microcirculation of mice subjected to sensitization with hen-egg white lysozyme. Among the principal compounds in IT, apigenin (1), luteolin (3), and luteolin 7-glucoside (4) showed significant allergy-preventive effects.

Allergy-preventive effects of Hibiscus mutabilis 'versicolor' and a novel allergy-preventive flavonoid glycoside.

Allergy-preventive activity was demonstrated for the MeOH extract (HM) of the petals of Hibiscus mutabilis L. 'versicolor' MAKINO in a continuing search for allergy-preventive substances from natural sources, using the in vivo assay method. This assay system uses monitoring of a decrease in the blood flow at the tail vein of mice subjected to hen egg-white lysozyme (HEL) sensitization. By bioassay-directed fractionation, a new flavonol triglycoside, quercetin 3-O-[beta-D-xylopyranosyl(1-->2)-alpha-L-rhamnopyranosyl(1-->6)]-beta-D-galactopyranoside (1: mutabiloside), was isolated, together with four known flavonols identified as quercetin 3-O-[beta-D-xylopyranosyl(1-->2)]-beta-D-galactopyranoside (2) and kaempferol 3-O-[beta-D-xylopyranosyl(1-->2)]-beta-D-galactopyranoside (3), quercetin (4) and hyperoside (5). The structure of the new flavonol 1 was elucidated by spectroscopic methods. Among these flavonol derivatives, compounds 1 and 2 showed significant allergy-preventive effects.

Involvement of inducible nitric oxide synthase in blood flow decrease in vein induced by hen-egg white lysozyme.

Our in vivo assay system developed to search for allergy-preventive substances, assesses the blood flow decrease in tail vein microcirculation of mice subjected to sensitization with hen-egg white lysozyme (HEL). The blood flow decrease appears to be regulated by various factors such as nitric oxide (NO), thromboxane (TX) A(2), prostacyclin (PGI(2)) and endothelin (ET)-1 together with cyclooxygenase (COX)-1, COX-2, inducible nitric oxide synthase (iNOS), and constitutive nitric oxide synthase (cNOS). In this study, we examined in detail the roles of iNOS in this assay system using an iNOS knockout (KO) mouse. We found that the blood flow decrease in the HEL-sensitized iNOS KO mice was slightly weaker than that in their wild type (WT) mice. This blood flow decrease was not affected by a selective COX-1 inhibitor, a selective COX-2 inhibitor and a PGI(2) agonist unlike the case of the WT mice. However, it was inhibited by a nonselective NOS inhibitor, a specific TXA(2) synthase inhibitor and a specific ET-1 receptor blocker as in the case of the WT mice. The present results indicate that the blood flow decrease occurs via two pathways; one is an iNOS-independent response involving TXA(2) and ET-1, and the other is an iNOS-dependent response involving COX-1, COX-2 and PGI(2). cNOS appears to play some roles in the blood flow decrease and iNOS acts as an exacerbation factor. Our method using HEL-sensitized should be useful for searching for agents that can prevent allergy via new mechanisms.

Allergy-preventive flavonoids from Xanthorrhoea hastilis.

Allergy-preventive activity was demonstrated for an extract of resins from Xanthorrhoea hastilis R. BR. in a search for allergy-preventive substances from natural sources. By bioassay-directed fractionation of this plant extract, a new flavanone, 3',5'-dihydroxy-7,4'-dimethoxyflavanone (1), and two new chalcones, 3,5,2'-trihydroxy-4,4'-dimethoxychalcone (2) and 5,2'-dihydroxy-3,4,4'-trimethoxychalcone (3), were isolated together with five known compounds, 5'-hydroxy-7,3',4'-trimethoxyflavanone (4), 3'-hydroxy-7,4'-dimethoxyflavanone (5), liquiritigenin 7-methyl ether (6), 4,2'-dihydroxy-4'-methoxychalcone (7) and sakuranetin (8). The structures of 1, 2 and 3 were elucidated by spectroscopic methods. All of these compounds showed allergy-preventive effects.

Allergy-preventive phenolic glycosides from Populus sieboldii.

Allergy-preventive activity was demonstrated for an extract of the bark of Populus sieboldii in a continuing search for allergy-preventive substances from natural sources. By bioassay-directed fractionation of this plant bark, two new phenolic glycosides, siebolside A {2-hydroxy-5-[(benzoyloxy)methyl]phenyl (6'-O-acetyl) beta-D-glucopyranoside} (1) and siebolside B {2-hydroxy-5-[(benzoyloxy)methyl]phenyl beta-D-glucopyranoside} (2), were isolated, together with three known compounds, salicin (3), sakuranin (4), and neosakuranin (5). The structures of 1 and 2 were elucidated by spectroscopic methods. Compounds 1-5 all showed allergy-preventive effects.

Garcinone B reduces prostaglandin E2 release and NF-kappaB-mediated transcription in C6 rat glioma cells.

In the course of our survey of natural compounds inhibiting prostaglandin E2 release and/or lipopolysaccharide (LPS)-induced transcriptional stimulation via NF-kappaB, a central regulator of inflammatory genes, from natural resources, we found garcinone B, a xanthone from callus tissue culture of Hypericum patulum, as a compound with such pharmacological activities, that is a derivative of gamma-mangostin which potently inhibits COX-1 and COX-2 activities to reduce PGE2 release from C6 rat glioma cells, and inhibits IKK activity to prevent NF-kappaB-dependent COX-2 gene transcription. Garcinone B, to a lesser extent, reduced A23187-induced increase in prostaglandin E2 release than gamma-mangostin and its structurally related compound, patulone, in C6 cells. This compound also prevented LPS-induced stimulation of NF-kappaB-dependent transcription. These results suggest that garcinone B becomes a unique pharmacological tool to investigate intracellular signaling pathways involved in inflammation.

Antianaphylactic and antipruritic effects of the flowers of Impatiens textori MIQ.

The anti-anaphylactic and anti-pruritic activities of a 35% EtOH extract (IT) of the flowers of Impatiens textori MIQ. were investigated by in vivo assay. IT and apigenin (1), apigenin 7-glucoside (2) and luteolin (3), principal compounds from IT, inhibited compound 48/80 (COM)-induced by blood pressure (BP) decrease, which was an immunoglobulin (Ig)E-independent anaphylaxis-like response. Compounds 1-3 all inhibited BP decrease induced by IgE-dependent anaphylaxis. Furthermore, IT also inhibited the blood flow (BF) decrease induced by antigen-induced anaphylaxis in actively sensitized mice. IT showed a significant inhibitory effect on scratching behavior induced by COM without a central depressant. IT also significantly inhibited platelet activating factor (PAF)- and serotonin (5-HT)-induced scratching behavior and mitigated protease (PA)-induced scratching behavior. These findings showed that the flowers of I. textori can be utilized as an anti-anaphylactic and anti-pruritic agent in addition to the traditional applications of this plant.

Development of an in vivo bioassay method for allergy-preventive substances using hen-egg white lysozyme (HEL)-induced blood flow decrease.

We discovered a phenomenon in which the blood flow in vein microcirculation markedly decreases in response to hen-egg white lysozyme (HEL)-sensitization without any change in blood pressure. Using this blood flow decrease as a guide, we developed an in vivo assay method to search for substances, which can prevent allergies. Antagonists of histamine, serotonin and platelet activating factor (PAF) did not affect the blood flow decrease in response to HEL-sensitization. On the other hand, cyclooxygenase (COX)-1, COX-2, thromboxane (TX) A(2), endothelin-1 (ET-1), prostacyclin (PGI(2)) and granulocytic elastase (GE) as well as nitric oxide (NO) from inducible NO synthase (iNOS) were involved in the blood flow decrease. Thus, these substances might injure vascular endothelial cells, and cause a decrease in blood flow in vein microcirculation. Our method can be used to search for preventive agents against allergies involving NO, COX-1, 2 and PGI(2). This is the first report to applying to an assay method the specific blood flow decrease to occur in the promotion stage of allergy.

Inhibitory effects of xanthones from guttiferae plants on PAF-induced hypotension in mice.

The inhibitory effects of 22 xanthones from three Guttiferae plants (Hypericum patulum, Calophyllum inophyllum and C. austroindium) on exogenous platelet activating factor (PAF)-induced hypotension were examined using a blood pressure monitoring in vivo assay method. Guanandin (2), caloxanthone E (3), 1,3,5,6-tetrahydroxy-2-isoprenylxanthone (8), 6-deoxyjacareubin (11) and patulone (18) showed strong inhibition of PAF-induced hypotension, with inhibitory effects of more than 60 %. Their ID50 values were greater than that of ginkgolide B (BN-52 021), a natural PAF-antagonist from the Ginkgo biloba.

Alpha-mangostin induces Ca2+-ATPase-dependent apoptosis via mitochondrial pathway in PC12 cells.

We investigated the cell death effects of eight xanthones on PC12 rat pheochromocytoma cells. Among these compounds, alpha-mangostin, from the fruit hull of Garcinia mangostana L., had the most potent effect with the EC(50) value of 4 microM. Alpha-mangostin-treated PC12 cells demonstrated typical apoptotic DNA fragmentation and caspase-3 cleavage (equivalent to activation). The flow cytometric analysis indicated that this compound induced apoptosis in time-and concentration-dependent manners. Alpha-mangostin showed the features of the mitochondrial apoptotic pathway such as mitochondrial membrane depolarization and cytochrome c release. Furthermore, alpha-mangostin inhibited the sarco(endo)plasmic reticulum Ca(2+)-ATPase markedly. There was a correlation between the Ca(2+)-ATPase inhibitory effects and the apoptotic effects of the xanthone derivatives. On the other hand, c-Jun NH(2)-terminal kinase (JNK/SAPK), one of the signaling molecules of endoplasmic reticulum (ER) stress, was activated with alpha-mangostin treatment. These results suggest that alpha-mangostin inhibits Ca(2+)-ATPase to cause apoptosis through the mitochondrial pathway.

Effects on blood pressure decrease in response to PAF of Impatiens textori MIQ.

A 35% EtOH extract of flowers of Impatiens textori MIQ. showed an inhibitory effect on blood pressure decrease in response to platelet activating factor (PAF) measured with a blood pressure monitoring system. Bioassay-guided fractionation of the 35% EtOH extract (IT) led to isolation of the flavones apigenin (1) and luteolin (3), which significantly inhibited blood pressure decrease in response to PAF. Their compounds and apigenin 7-glucoside (2), chrysoeriol (4), quercetin (5), quercetin 3-glucoside (6), kaempferol (7), kaempferol 3-glucoside (8) and kaempferol 3-rhamnosyldiglucoside (9) were also isolated from the flowers of I. textori for the first time. This study revealed that the flowers of I. textori might be a possible anti-allergy agent.

Antipruritic effects of the fruits of Chaenomeles sinensis.

A 35% EtOH extract of the fruits of Chaenomeles sinensis, long utilized as a folk medicine for cough, significantly inhibited the pruritogenic agent compound 48/80 (COM)-induced scratching behavior in mice. Antipruritic activity-guided fractionation and purification yielded active quercetin, apigenin, and catechin derivatives, which exhibited significant inhibitory effects on COM-induced scratching behavior. To the best of our knowledge, apigenin (5), apigenin 7-glucronide (6), and apigenin 4'-methoxy-7-glucronide (acacetin 7-glucronide) (7) were isolated from the fruits of C. sinensis for the first time. The active fraction and these compounds also inhibited serotonin-, platelet activating factor-, and prostaglandin E(2)-induced scratching behavior, but did not inhibit histamine-induced scratching behavior or locomotive behavior. This study also showed that the fruits of C. sinensis could be used to treat allergic itching sensation.

Effects of conjugated linoleic acid on anaphylaxis and allergic pruritus.

The effects of conjugated linoleic acid (CLA) against anaphylaxis and allergic pruritus were investigated using a in vivo assay. Inhibitory effects of CLA were observed on the immediate (type 1) hypersensitivity reaction, with CLA significantly suppressing the decrease in blood pressure (BP) and blood flow (BF) induced by the hen egg-white lysozyme (HEL)-anaphylactic reaction in ddY mice. After oral administration, CLA showed antipruritic activity, with significant inhibition of scratching behavior induced by compound 48/80 (COM), a histamine-release agent. When painted onto the skin, CLA also inhibited COM, platelet-activating factor, and protease-induced scratching behavior, and COM-induced vasodilation of the skin. CLA offers promise as a drug for the treatment of allergic and inflammatory pruritus not only as an oral but also a topical agent. The present findings demonstrate that CLA can be effective for the prevention and treatment of allergic disease with severe pruritus.

Anti-inflammatory and anti-allergic activities of hydroxylamine and related compounds.

The anti-inflammatory activities of several novel oximes and O-acyl oximes that we synthesized have been reported based on carrageenan-induced rat foot-pad swelling assay and histamine-induced rat vascular permeability assay. A cyclooxygenase (COX)-1 inhibitory effect has also been reported for 4'-piperidinoacetophenone and 4'-morpholinoacetophenone oximes and their O-acyl derivatives. To further search for more effective non-steroidal anti-inflammatory or anti-allergic drugs, 1-hydroxylamino-1-(4'-piperidinophenyl) ethane (P-HA) and 1-hydroxylamino-1-(4'-morpholinophenyl) ethane (M-HA) were synthesized from the corresponding oximes with sodium cyanoborohydride, and N,O-diacetyl hydroxylamines (P-HA-Ac and M-HA-Ac) were prepared from these hydroxylamines using acetyl chloride. These hydroxylamines and N,O-diacetyl hydroxylamines clearly exhibited inhibitory effects on mouse carrageenan-induced foot-pad swelling induced by oral administration (150, 37.5 mg/kg). An oral dose of P-HA-Ac (150 mg/kg) significantly inhibited the mouse anaphylactic reaction to ovalbumin measured by the abdominal wall (AW) method. Percutaneous administration of P-HA and M-HA significantly inhibited 2,4-dinitrofluorobenzene (DNFB)-induced contact hypersensitivity reaction (type IV) in mice at a dose of 0.5 and 0.1 mg/ear, respectively. All tested hydroxylamines and N,O-diacetyl hydroxylamines clearly inhibited both COX-1 and COX-2 enzyme activities with IC(50) values of 1.9-28.7 and 1.6-2.9 micro M against COX-1 and COX-2, respectively. Hydroxylamines (P-HA and M-HA) also showed a 5-lipoxygenase inhibitory effect.