Evodia rutaecarpa and Three Major Alkaloids Abrogate Influenza A Virus (H1N1)-Induced Chemokines Production and Cell Migration.

Evodia rutaecarpa is commonly used as an anti-inflammatory herbal remedy in traditional Chinese medicine. In this study, the ethanol extract of E. rutaecarpa (ER) and three major quinazoline alkaloids dehydroevodiamine (DeHE), evodiamine (Evo) and rutaecarpine (Rut), isolated from ER were employed to study their inhibitory effects against influenza A virus (H1N1)-induced chemokines production in A549 lung epithelial cells as well as on chemokines-evoked cell recruitment in HL-60-differentiated macrophages. The results showed that ER was a potent inhibitor of RANTES secretion by H1N1-inoculated A549 cells (IC(50): 1.9 ± 0.4 µg ml(-1)). Three alkaloids, although to differing extents, all concentration dependent, inhibited H1N1-induced RANTES production with Evo consistently being the most potent among these active components. ER also moderately and significantly inhibited H1N1-stimulated MCP-1 production in A549 cells. This was mimicked by Evo and Rut, but not DeHE. In the macrophage recruitment assay, both RANTES and MCP-1 markedly evoked cell migration and this phenomenon was significantly suppressed by ER. Evo and Rut, but not DeHE, also had the ability to inhibit cell migration toward RANTES and MCP-1, respectively. In summary, three major alkaloids displayed different potentials for inhibiting chemokines secretion and subsequently cell migration, which could partially explain the activity of ER. As an effective agent to suppress H1N1-induced chemokines production and block chemokine-attracted leukocytes recruitment, E. rutaecarpa and its active components may be useful in influenza virus infection-related inflammatory disorders.

Anti-inflammatory effects and mechanisms of the ethanol extract of Evodia rutaecarpa and its bioactive components on neutrophils and microglial cells.

Evodia rutaecarpa is commonly used as an anti-inflammatory drug in traditional Chinese medicine. We previously identified four bioactive compounds (dehydroevodiamine (I), evodiamine (II), rutaecarpine (III), and synephrine (IV)) from the ethanol extract of E. rutaecarpa, but their effects and mechanism(s) of action remain unclear. To study the anti-inflammatory potential and the possible underlying mechanism(s), their effects on phorbol-12-myristate-13-acetate (PMA)- and N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced reactive oxygen species production in neutrophils was studied, as well as lipopolysaccharide (LPS)-induced nitric oxide (NO) production and inducible NO synthetase (iNOS) expression in microglial cells. The ethanol extract of E. rutaecarpa displayed potent antioxidative effects against both PMA- and fMLP-induced reactive oxygen species production in neutrophils (with IC50 values of around 2.7-3.3 microg/ml). Although less potent than the ethanol extract of E. rutaecarpa, compounds I-IV all concentration-dependently inhibited PMA- and fMLP-induced reactive oxygen species production, with compound IV consistently being the most potent agent among these active components. The antioxidative effects of the ethanol extract of E. rutaecarpa and these compounds were partially due to inhibition (10%-33%) of NADPH oxidase activity, a predominant reactive oxygen species-producing enzyme in neutrophils, and to a minor extent to their direct radical-scavenging properties. The ethanol extract of E. rutaecarpa also inhibited LPS-induced NO production (with an IC50 of around 0.8 microg/ml) and iNOS upregulation in microglial cells that was partially mimicked by compounds I, II, and III, but not compound IV. Our results suggest that the ethanol extract of E. rutaecarpa and its four bioactive components all exhibited anti-inflammatory activities which could be partially explained by their different potentials for inhibiting NADPH oxidase-dependent reactive oxygen species and/or iNOS-dependent NO production in activated inflammatory cells.

The effect of medicinal plants used in Chinese folk medicine on RANTES secretion by virus-infected human epithelial cells.

The accumulation of inflammatory cells in the infective sites has been reported to play a crucial role in the progression of chronic inflammation and multiple sclerosis after viral infection. In the present study, nine ethanol extracts of Forsythia suspensa Vahl. (Oleaceae), Lonicera japonica Thunb. (Caprifoliaceae), Isatis indigotica Fort. (Cruciferae), Strobilanthes cusia (Ness.) O. Kuntze (Acanthaceae), Astragalus membranaceus (Fisch.) Bge. (Leguminosae), Hedysarum polybotrys Hand.-Mazz. (Leguminosae), Andrographis paniculata (Burm. f.) Ness. (Acanthaceae), Glycyrrhiza uralensis Fischer. (Leguminosae) and Ligusticum wallichii Franch. (Umbelliferae), medicinal plants traditionally used in China for treating conditions likely to be associated with inflammation and viral infection, were screened for their effect on RANTES secretion by influenza A virus (H1N1)-infected human bronchial epithelial cells (A549). With exception of Lonicera japonica, Isatis indigotica, Astragalus membranaceus and Hedysarum polybotrys, all plants tested at concentration of 200 microg/ml possessed more than 50% suppressing effect on RANTES secretion by H1N1-infected A549 bronchial epithelial cells. Among the plants tested, Andrographis paniculata showed the most promising property to inhibit RANTES secretion with an IC(50) of 1.2 +/- 0.4 microg/ml while the next two were Glycyrrhiza uralensis and Forsythia suspensa (IC(50) ranging from 35 to 48 microg/ml).

Chemical and biological comparisons on Evodia with two related species of different locations and conditions.

Evodia rutaecarpa (ER) and Tetradium glabrifolium (TG) are closely related species collected from different locations, with processed versus unprocessed and fresh versus 1-year-old samples. The purpose of this study is to determine the variability of their bioactive constituents; evodiamine, dehydroevodiamine, rutaecarpine and synephrine--as well as their relaxing effects on an isolated rat aortas and uterus using the extracts of the test specimens. The vasorelaxation was greater in ER from Taiwan than from China in spite of lower levels of the relaxing alkaloids evodiamine, dehydroevodiamine and rutaecarpine. On the other hand, the uterine relaxation of ER from China was better than the one from Taiwan, even though constricting synephrine was only contained in Chinese ER. After processing, the relaxation of ER from China in the uterus was increased while the vasorelaxation remained unchanged. Conversely, TG from Wu-ling contained more relaxing alkaloids than that from Lee Mountain. However, the relaxation in both the uterus and the aorta was less in TG from Wu-ling. After 1 year of storage, the vasorelaxation of TG from Lee Mountain was not changed. Taken together, a significant finding in the present study is the lack of correction between chemical composition and relaxing activities. This strongly supports our assumption that biological function evaluations, instead of chemical standardization, is the more adequate way of showing meaningful consistency of natural preparations.

Dual regulatory effect of plant extracts of Forsythia suspense on RANTES and MCP-1 secretion in influenza A virus-infected human bronchial epithelial cells.

In this study, we investigated the effects of 95% ethanol (FS-t1), 50% ethanol (FS-t2) and water (FS-w) extracts of Forsythia suspense Vahl (Oleaceae) on the production of regulated on activation, normal T cell expressed and secreted (RANTES) and macrophage chemotactic protein-1 (MCP-1) by influenza A virus (H1N1)-infected human bronchial epithelial cell line A549. Virus infection evoked a markedly enhanced production of RANTES from basal 16 +/- 4 to 1307 +/- 294 pg/ml after 72 h inoculation. At the non-cytotoxic doses (20, 100 and 200 microg/ml), FS-t1, FS-t2 and FS-w exhibited a consistent inhibitory effect on virus-stimulated RANTES secretion in a dose-dependent manner wilh IC(50) of 42 +/- 6, 117 +/- 15 and 232 +/- 28 microg/ml, respectively. H1N1 also stimulated MCP-1 production in A549 cells, however to a less degree, from basal 133 +/- 21 to 391 +/- 98 pg/ml after 72 h viral inoculation. The effects of three extracts on MCP-1 secretion were more complex. FS-t1 displayed both positive and negative effect on virus-stimulated MCP-1 production dependent on the concentrations used. On the other hand, FS-t2 increased virus-induced MCP-1 secretion by 1.4-3.3 times while the third fraction FO-w increased by 2.6-3.7 times. These results suggested that Forsythia suspense consisted of both negative and positive regulatory components on RANTES and MCP-1 secretion in H1N1-infected A549 cells, respectively.

Modulation of drug-metabolizing enzymes by extracts of a herbal medicine Evodia rutaecarpa in C57BL/6J mice.

Evodia rutaecarpa is a traditional Chinese medicine used for the treatment of gastrointestinal disorders and headache. To assess the possible drug interactions, effects of methanol and aqueous extracts of E. rutaecarpa on drug-metabolizing enzymes, cytochrome P450 (CYP), UDP-glucuronosyl transferase (UGT), and glutathione S-transferase (GST) were studied in C57BL/6J mice. Treatment of mice with methanol extract by gastrogavage caused a dose-dependent increase of liver microsomal 7-ethoxyresorufin O-deethylation (EROD) activity. In liver, methanol extract at 2 g/kg caused 47%, 7-, 8-, 4-fold, 81% and 26% increases of benzo(a)pyrene hydroxylation (AHH), EROD, 7-methoxyresorufin O-demethylation (MROD), 7-ethoxycoumarin O-deethylation (ECOD), benzphetamine N-demethylation, and N-nitrosodimethylamine N-demethylation activities, respectively. Aqueous extract at 2 g/kg caused 68%, 2-fold, and 83% increases of EROD, MROD, and ECOD activities, respectively. For conjugation activities, methanol extract elevated UGT and GST activities. Aqueous extract elevated UGT activity without affecting GST activity. Immunoblot analyses showed that methanol extract increased the levels of CYP1A1, CYP1A2, CYP2B-, and GSTYb-immunoreactive proteins. Aqueous extract increased CYP1A2 protein level. In kidney, both extracts had no effects on AHH, ECOD, UGT, and GST activities. Three major bioactive alkaloids rutaecarpine, evodiamine, and dehydroevodiamine were present in both extracts. These alkaloids at 25 mg/kg increased hepatic EROD activity. These results demonstrated that E. rutaecarpa methanol and aqueous extracts could affect drug-metabolizing enzyme activities. Rutaecarpine, evodiamine, and dehydroevodiamine contributed at least in part to the increase of hepatic EROD activity by extracts of E. rutaecarpa. Thus, caution should be paid to the possible drug interactions of E. rutaecarpa and CYP substrates.

Evodia rutaecarpa protects against circulation failure and organ dysfunction in endotoxaemic rats through modulating nitric oxide release.

Using a rat model of septic shock we studied the effects of Evodia rutaecarpa, a Chinese herbal medicine with antimicrobial and anti-inflammatory activity, on haemodynamic parameters, biochemical markers of organ function and nitric oxide (NO) production. Anaesthetized rats challenged with a high dosage of endotoxin (Escherichia coli lipopolysaccharide; LPS; 50 mg kg(-1), i.v.) for 6 h showed a severe decrease in mean arterial pressure. This was accompanied by delayed bradycardia, vascular hyporeactivity to phenylephrine and increase in plasma levels of lactate dehydrogenase, aspartate aminotransferase, bilirubin and creatinine, as well as NOx (NO2- plus NO3-). Pretreatment with ethanol extract of E. rutaecarpa (25, 50 and 100 mg kg(-1), i.v.), 1 h before LPS, dose-dependently prevented the circulation failure, vascular hyporeactivity to phenylephrine, prevented liver dysfunction and reduced the NOx over-production in plasma in endotoxaemic rats. A selective inducible NO-synthase (iNOS) inhibitor, aminoguanidine (15 mg kg(-1), i.v.), also effectively ameliorated the above pathophysiological phenomenon associated with endotoxaemia so that the normal condition was approached. Endotoxaemia for 6 h resulted in a significant increase in iNOS activity in the liver homogenate, which was attenuated significantly by E. rutaecarpa pretreatment. In summary, E. rutaecarpa, at the dosages used, exerted these beneficial effects probably through inhibition of iNOS activity and subsequent modulation of the release of NO. These significant results may offer E. rutaecarpa as a candidate for the treatment of this model of endotoxaemia.

Anti-inflammatory ergostanes from the basidiomata of Antrodia salmonea.

Three new anti-oxidative ergostanes, methyl antcinate L (1), antcin M (2), and methyl antcinate K (3), together with nine additional known compounds, 3-ketodehydrosulphurenic acid, sulphurenic acid, dehydrosulphurenic acid, 3beta,15alpha-dihydroxylanosta-7,9(11),24-trien-21-oic acid, zhankuic acid A, zhankuic acid B, zhankuic acid C, antcin C, and antcin K were isolated from the basidiomata of Antrodia salmonea, a newly identified species of Antrodia (Polyporaceae) in Taiwan. These three new compounds were identified as methyl 3alpha,7alpha,12alpha-trihydroxy-4alpha-methylergosta-8,24(29)-dien-11-on-26-oate (1), 3alpha,12alpha-dihydroxy-4alpha-methylergosta-8,24(29)-dien-11-on-26-oic acid (2), and methyl 3alpha,4beta,7beta-trihydroxy-4alpha-methylergosta-8,24(29)-dien-11-on-26-oate (3) by spectroscopic analysis. We studied their antioxidative potential on the production of reactive oxygen species and nitric oxide (NO) in neutrophils and microglial cells, respectively. Compounds 1-3 displayed potent antioxidative activity with IC50 values of around 2.0-8.8 microM that was partially due to inhibition (6-67%) of NADPH oxidase activity but not through direct radical-scavenging properties. Compounds 1-3 also inhibited NO production with IC50 values of around 1.7-16.5 microM and were more potent than a non-specific NOS inhibitor. We conclude that these three new compounds 1, 2, and 3 exhibit anti-inflammatory activities in activated inflammatory cells.

Antidepressant principles of the roots of Polygala tenuifolia.

[(125)I]RTI-55-membrane binding assay-guided fractionation and separation of a water-soluble extract of the roots of Polygala tenuifolia gave five new oligosaccharide derivatives, polygalatenosides A-E (1-5). The structures of these new oligosaccharides were established on the basis of spectroscopic evidence. Polygalatenosides A and B (1 and 2) showed significant inhibitory activity, with IC(50) values of 30.0 and 6.04 microM, respectively, in this membrane binding assay and acted as norepinephrine reuptake inhibitors through blocking norepinephrine transport.

Laxifolone A suppresses LPS/IFN-gamma-induced NO synthesis by attenuating NF-kappaB translocation: role of NF-kappaB p105 level.

Laxifolone A is a triterpene isolated from Euonymus laxiflorus Champ. Exposure of RAW264.7 macrophages to laxifolone A concentration-dependently suppressed lipopolysaccharide/interferon-gamma (LPS/IFN-gamma)-induced nitrite production (IC50 = 0.37 +/- 0.05 microM), inducible NO synthase (iNOS) protein, and iNOS mRNA expression. Translocation of nuclear factor-kappaB (NF-kappaB) into the nucleus with subsequent activation of iNOS gene transcription is essential in NO signaling. Western blot analysis indicated that the cytosolic NF-kappaB/p65 was obviously decreased after LPS/IFN-gamma stimulation for 30 min and this phenomenon could be reversed by laxifolone A. Similarly, a time-related NF-kappaB/p65 nuclear translocation induced by LPS/IFN-gamma was diminished in the presence of laxifolone A. However, laxifolone A failed to interfere in LPS/IFN-gamma-evoked IkappaB degradation. Our results also showed that LPS/IFN-gamma-stimulation resulted in the degradation of NF-kappaB p105, the NF-kappaB precursor, and laxifolone A treatment significantly counteracted this effect. Furthermore, laxifolone A itself was able to enhance NF-kappaB p105 protein expression. In summary, these results suggest that inhibition of NF-kappaB p105 degradation in cytoplasm may participate in the abrogation of LPS/IFN-gamma-induced NF-kappaB translocation and subsequent NO synthesis by laxifolone A.