Release Of Beta-glucuronidase Research Articles

Chemical constituents from Machilus wangchiana

Twenty-one compounds were isolated from an ethanol extract of Machilus wangchiana by a combination of various chromatographic techniques including column chromatography over silica gel and Sephadex LH-20 and reversed-phase HPLC. Their structures were identified by spectroscopic data analysis including optical rotation, UV, IR, MS, and NMR data. The compounds are categorized as eight butanolides (1-8), eight lignans (9-16), and five terpenoids (17-21). Compound 16 is a new natural product with an uncommon heptanorlignan skeleton. Meanwhile, the unique Ginkgo biloba (maidenhair) metabolites ginkgolides A (19) and ginkgolides B (20) were obtained from this material. In the preliminary assays, compound 5 showed selective inhibitory activities against human stomach cancer cells (BGC-823) and ovary cancer cells (A2780) with IC50 values of 0.13 x 10(-6) and 2.66 x 10(-6) mol x L(-1), respectively. Compounds 8 and 9, at 1 x 10(-5) mol x L(-1), showed inhibitory activities against the release of beta-glucuronidase of the polymorphous nuclear leukocytes induced by platelet activating factor (PAF), with inhibition rates of 60.0% and 54.2%.

Synthesis and anti-inflammatory effects of xanthone derivatives.

Eighteen synthetic xanthone derivatives were tested for their inhibitory effects on the activation of mast cells and neutrophils. 1,3- and 3,5-Dihydroxyxanthone showed strong inhibitory effects on the release of beta-glucuronidase and histamine from rat peritoneal mast cells stimulated with compound 48/80. 1,6-Dihydroxyxanthone and 1,3,8-trihydroxyxanthone showed strong inhibitory effects on the release of beta-glucuronidase, and beta-glucuronidase and lysozyme, respectively, from rat neutrophils stimulated with formyl-Met-Leu-Phe (fMLP). 1,3- and 1,6-Dihydroxyxanthone, 1,3,7-trihydroxyxanthone, and 1,3,5,6-, 2,3,6,7-, and 3,4,5,6-tetrahydroxyxanthone showed potent inhibitory effects on superoxide formation of rat neutrophils stimulated with fMLP. 1,6- and 3,5-Dihydroxyxanthone showed remarkable inhibitory effects on hind-paw oedema induced by polymyxin B in normal as well as in adrenalectomized mice. These data indicated that the anti-inflammatory effect of these compounds is mediated through the suppression of chemical mediators released from mast cell and neutrophil degranulation.

Aromatic constituents of Heteroplexis micocephal and their bioactivities

To investigate the chemical constituents of Heteroplexis micocephal and their bioactivities. The constituents were isolated by using a combination of various chromatographic techniques including column chromatography over macroporous adsorbent resin, silica gel, Pharmadex LH-20, and C-18, as well as reversed-phase HPLC. Structures of the isolates were identified by spectroscopic data analysis. In vitro cytotoxic, HIV-1 replication, neuroprotective, and anti-inflammatory activities were screened by using cell-based models. Thirty-one compounds were obtained. Twelve of them are phenylpropanols, and the structures were elucidated as (+)-(7S,8R)-guaiacylglycerol (1), ferulic acid (2), cinnamate methyl ester (3), 1-eicosanyl 3,4-dihydroxycinnamate (4), morinin B (5), sinapyl diangelate (6), chlorogenic acid (7), 4-O-caffeoylquinic acid (8), 5-O-caffeoylquinic acid (9), 5-O-caffeoylquinic acid methyl ester (10), 1,5-di-O-caffeoylquinic acid (11) and 4,5-di-O-caffeoylquinic acid methyl ester (12). Three lignans, (+)-pinoresinol (13), prinsepiol (14) and (+)-pinoresinol-O-beta-D-glucopyranoside (15). Four acetophenones, 2,4-diacetylanisole (16), espeleton (17), viscidone (18) and 12-hydroxytremetone-12-O-beta-D-glucopyranoside (19). Nine flavones, isosakuranetin (20), hesperetin (21), 3-methoxy-5,7,3',4'-tetrahydroxyflavone (22), acacetin (23), 5-hydroxy-7,4'- dimethoxyflavone (24), 7-methoxy-4',5, 6-trihydroxyflavone (25), 3,3'-dimethylquercetin (26), kaempferol 3-O-rutinoside (27), rutin (28). And three coumarins scopoletin (29), umbelliferone (30) and ayapin (31). Compound 6 and 22 showed selective cytotoxicities against a human stomach cancer cell line(BGC-823) and a human lung cancer cell line (A549) with IC50 values of 3.74 x 10(-5) and 7.17 x 10(-5) mol L(-1), respectively. In addition, Compound 6 showed a potent activity inhibiting HIV-1 replication with an IC50 value of 4.04 x 10(-6) mol L(-1), while 22 showed neuroprotective activity Against the MPP+ induced PC12-syn cell damage, with a relative protection ratio of 105.2% (P < 0.01) at a concentration of 10(-5) mol L(-1). Compound 26 and 31 showed inhibitory activities against the release of beta-glucuronidase of the polymorphous nuclear leukocytes induced by platelet activating factor (PAF), with inhibitory rates of 75.6% (P < 0.001) and 53. 9% (P < 0.01), respectively. Compounds 1-31 were obtained from the genus Heteroplexis for the first time. Compound 6 and 22 possessed selective cytotoxicities against human cancer cell lines BGC-823 and A549, respectively. In addition, Compound 6 showed a potent activity inhibiting HIV-1 replication while 22 showed neuroprotective activity against the MPP+ induced PC12-syn cell damage. Compound 26 and 31 were potent anti-inflammatory agents.

Phenolic constituents from stem bark of Morus wittiorum and their anti-inflammation and cytotoxicity

To search for the chemical constituents possessing anti-inflammatory or cytotoxic activities from plants, Morus wittiorum was investigated for the first time. The stem bark of M. wittiorum was extracted with 95% EtOH. The EtOH extract was fractionationed on silica gel by eluting with petroleum ehter, CHCl3 and EtOAc successively. The further isolation and purification of the EtOAc fraction of 95% EtOH extract was performed by various column chromatography such as silica gel, Sephadex LH-20, RP-C18 column chromatography and so on. The structures of compounds were determined on the basis of spectral analysis such as NMR, MS etc. As a result, nine compounds were isolated including six flavonoids and three stilbenoids and elucidated as quercetin (1), 5, 7, 3', 4'-tetrahydroxy-3-methoxyflavone (2), norartocarpanone (3), dihydrokaempferol (4), euchrenone a7 (5), morachalcone A (6), resveratrol (7), oxyresveratrol (8), 4'-prenyloxyresveratrol (9). Compounds 1-9 were isolated from this plant for the first time, among which compounds 1-8 were evaluated for their anti-inflammatory and cytotoxic activities, respectively. Wherein compounds 6 and 8 showed inhibition to the release of beta-glucuronidase from rat polymorphonuclear leukocyte (PMNs) induced by platelet activating factor (PAF) at a concentration of 10(-5) mol x L(-1). The inhibitory ratios were 76.8%, 94.2% individually. Compounds 2 and 8 exhibited selective cytotoxicity agaist human ovarian cancer (A2780) and human gastric cancer (BGC-823) with IC50 values as 0.66, 1.31 micromol x L(-1) individually.

Thiodiketopiperazines Produced by the Endophytic Fungus Epicoccum nigrum

Thirteen new thiodiketopiperazines, epicoccin I (1), ent-epicoccin G (2), and epicoccins J-T (3-13), together with six known diketopiperazines (14-19), have been isolated from the endophytic fungus Epicoccum nigrum. The structures of 1, 2, and 10 were confirmed by X-ray crystallography, and the absolute configurations of 2, 4, 6, and 8 were assigned using Moshers' method. Compounds 2, 6, 12, and 17 showed potent activities in vitro against the release of beta-glucuronidase in rat polymorphonuclear leukocytes induced by platelet-activating factor, with IC(50) values of 3.07, 4.16, 4.95, and 1.98 microM, respectively. None of the 19 compounds exhibited detectable cytotoxic activities toward six tumor cell lines (A549, Be-l7402, BGC-823, HCT-8, HCT-116, and A2780) in the MTT assay.

Chemical constituents of stems and branches of Adina polycephala

To investigate chemical constituents of the stems and branches of Adina polycephala and their pharmacological activities. The constituents were isolated by a combination of various chromatographic techniques including column chromatography on silica gel, Sephadex LH-20, and C-18, as well as reversed-phase HPLC. Structures of the isolates were identified by spectroscopic data analysis. In vitro cytotoxic, anti-inflammatory, anti-oxidant, anti-HIV, neuroprotective and anti-diabetic activities were screened by using cell-based models. Twenty-eight constituents were isolated. Their structures were identified as clemochinenoside B (1), kelampayoside A (2), osmanthuside H (3), 4-hydroxy-3-methoxyphenol-beta-D-[6-O-(4-hydroxy-3,5-dimethoxylbenzoate)]-glucopyranoside (4), and syringic acid beta-D-glucopyranosyl ester (5). Ten iridoidal glycosides: geniposidic acid (6), geniposide (7), 6beta-hydroxygeniposide (8), 6beta-hydroxygeniposide (9), ixoside (10), ixoside 11-methyl ester (11), 11-methyl forsythide (12), 7beta-hydroxysplendoside (13), gardoside (14) and mussaenosidic acid (15), (+) -pinoresinol (16), (+) -medioresinol (17), (+) -syringaresinol (18), (-)-lariciresinol (19), evofolin-B (20), alpha-hydroxyacetovaillone (21), syringic acid (22), vanillin (23), 3, 4, 5-trimethoxyphenol (24), and 2,6-dimethoxy-1, 4-benzoquinone (25), beta-sitosterol (26), mannitol (27), and daucosterol (28). At a concentration of 1.0 x 10(-5) mol x L(-1), these compounds were inactive in the assays, including cytotoxicity against human tumor cell lines (HCT-8, Bel-7402, BGC-823, A549 and A2780), anti-inflammatory activity against the release of beta-glucuronidase in rat polymorphonuclear leukocytes (PMNs) induced by platelet-activating factor (PAF), antioxidant activity in Fe(2+)-cystine-induced rat liver microsomal lipid peroxidation, anti-HIV activity against HIV-1 replication, neuroprotective activity against serum deprivation or glutamate induced neurotoxicity in cultures of PC12 cells, and the inhibitory activity against protein tyrosine phosphatase 1B (PTP1B). Compounds 1-20 were obtained from the genus Adina for the first time. The 13C-NMR data of compounds 10 and 11 were reassigned. A further evaluation of pharmacological activity of these compounds is expected.

Synthesis and anti-inflammatory effect of chalcones.

The process of degranulation of mast cells and neutrophils contributes to inflammatory disorders. Activation of microglial cells and macrophages is believed to be involved in inflammatory, infectious and degenerative diseases of the CNS. Combining the potent inhibition of chemical mediators released by the degranulation of mast cells or neutrophils and from the activated microglial cells or macrophages, would lead to a promising anti-inflammatory agent for the treatment of peripheral and central inflammation. A series of chalcone derivatives have been reported to have potent anti-inflammatory activity. In an effort to continually develop potent anti-inflammatory agents, novel series of chalcones, 2'-hydroxy- and 2',5'-dihydroxychalcones were synthesized and their inhibitory effects on the activation of mast cells, neutrophils, microglial cells and macrophages were evaluated in-vitro. The chalcones were prepared by Claisen-Schmidt condensation of appropriate acetophenones with an appropriate aromatic aldehyde. The alkoxychalcones were prepared with appropriate hydroxychalcones and alkyl iodide and the dihydroxychalcones were prepared by hydrogenation of an appropriate chalcone with Pd/C. Almost all of the hydroxychalcones exhibited potent inhibitory effects on the release of beta-glucuronidase and lysozyme from rat neutrophils stimulated with formyl-Met-Leu-Phe/cytochalasin B (fMLP/CB). Of the hydroxychalcones, compound 1 was the most potent inhibitor of the release of beta-glucuronidase (IC50=1.6+/-0.2 microM) and lysozyme (IC50=1.4+/-0.2 microM) from rat neutrophils stimulated with fMLP/CB. Almost all of the 2',5'-dialkoxychalcones exhibited potent inhibitory effects on nitric oxide (NO) formation from murine microglial cell lines N9 stimulated with lipopolysaccharide (LPS). Of these, compound 11 showed the greatest effect (IC50=0.7+/-0.06 microM). The present results demonstrated that most of the chalcone derivatives have an anti-inflammatory effect. The inhibitory effects of dialkoxychalcones, 10-12 on inflammation are probably not due to the inhibition of mast cells and neutrophil degranulation, but are mediated through the suppression of NO formation from N9 cells.

Terpenoids of Heteroplexis micocephala and their bioactivities

To investigate the chemical constituents of Heteroplexis nicocephala. The constituents were isolated by using a combination of various chromatographic techniques including column chromatography over silica gel, Pharmadex LH-20, and C-18, as well as reversed-phase HPLC. Structures of the isolates were identified by spectroscopic data analysis. In vitro cytotoxic, neuroprotective, and anti-inflammatory activities were screened by using cell-based models. Seventeen terpenoids were isolated. The structures were identified as two monoterpenoids: (-)-bomyl ferulate(1) and loliolide(2). Seven sesquiterpenoids: 1beta-hydroxy-alpha-cyperone(3) , alpha-rotunol(4), 10alpha-hydroxycadin-4-en-15-al (5), 1-epi-10beta-hydroxycadin-4-en-15-al(6), 10alpha-hydroxyisodauc-3-en-15-al(7), germacrene B(8), and mandassidione(9). Five diterpenoids: 12-epi-bacchotricuneatin A(10), 1-hydroxy-12-epi-bacchotricuneatin A(11), cleroinermin(12), desoxyarticulin(13), and anhydroolearin(14). And three triterpenoids: friedelin (15), ursolic acid(16), and obtusalin(17). In the in vitro assays, 1 showed selective cytotoxic activity against BGC-823, with an IC50 value of 8.00 x 10(-5) mol x L(-1). At a concentration of 1 x 10(-5) mol x L(-1), 12 showed neuroprotective activity against MPP+ induced PC12-syn cell damage, with a relative cell proliferation rate of 104.32% (P < 0.001). 2 exhibited inhibitory activity against the release of beta-glucuronidase from the polymorphous nuclear leukocytes induced by PAF, with an inhibitory rate of 52.7% (P < 0.05) at the same concentration. Compounds 1-17 were obtained from the genus Heteroplexis for the first time. 1 showed selective cytotoxic activity against human gastric cancer cell lines (BGC-823), 12 and 2 showed potent neuroprotective and anti-inflammatory activities, respectively.

New Anti-inflammatory 4-Hydroxyisoflavans from Solanum lyratum

Three new 4-hydroxyisoflavans, named lyratin A (1), lyratin B (2) and lyratin C (3), along with a known compound, 4,7,2'trihydroxy-4'-methoxyisoflavan (4), were isolated from the whole plant of Solanum lyratum. Their structures were established by means of detailed physical data analyses. In vitro, four compounds showed anti-inflammatory activities with inhibitory ratios of release of beta-glucuronidase from polymorphonuclear leukocytes of rats in the range of 30.3-38.6% at 10 microM.

New Alkaloids fromForsythia suspensaand their Anti-Inflammatory Activities

Two new alkaloids, named suspensine A (1) and (-)-7'-O-methylegenine (2), along with two known alkaloids, (-)-egenine (3) and (-)-bicuculline (4), were isolated from the ethanolic extract of the fruits of Forsythia suspensa. Their structures were established by means of chemical methods and spectroscopic analyses. In vitro, four alkaloids showed anti-inflammatory activities, with inhibition rates of release of beta-glucuronidase from polymorphonuclear leukocytes of rats being in the range 34.8%-39.6% at 10 microM.

Glycosides from the Bark of Adina polycephala

Four new dimeric phenolic glycosides (1- 4), a new iridoid diglycoside (5), and 15 known glycosides have been isolated from an ethanolic extract of the bark of Adina polycephala. Their structures were determined by spectroscopic and chemical methods. Compounds 1, 3, and 5 showed in vitro inhibitory activity against the release of beta-glucuronidase in rat polymorphonuclear leukocytes induced by platelet-activating factor.

Optimization of pulsed electromagnetic field therapy for management of arthritis in rats

Studies were undertaken to find out the effects of low frequency pulsed electromagnetic field (PEMF) in adjuvant induced arthritis (AIA) in rats, a widely used model for screening potential therapies for rheumatoid arthritis (RA). AIA was induced by an intradermal injection of a suspension of heat killed Mycobacterium tuberculosis (500 mug/0.1 ml) into the right hind paw of male Wistar rats. This resulted in swelling, loss of body weight, increase in paw volume as well as the activity of lysosomal enzymes viz., acid phosphatase, cathepsin D, and beta-glucuronidase and significant radiological and histological changes. PEMF therapy for arthritis involved optimization of three significant factors, viz., frequency, intensity, and duration; and the waveform used is sinusoidal. The use of factorial design in lieu of conventional method resulted in the development of an ideal combination of these factors. PEMF was applied using a Fransleau-Braunbeck coil system. A magnetic field of 5 Hz x 4 muT x 90 min was found to be optimal in lowering the paw edema volume and decreasing the activity of lysosomal enzymes. Soft tissue swelling was shown to be reduced as evidenced by radiology. Histological studies confirmed reduction in inflammatory cells infiltration, hyperplasia, and hypertrophy of cells lining synovial membrane. PEMF was also shown to have a membrane stabilizing action by significantly inhibiting the rate of release of beta-glucuronidase from lysosomal rich and sub-cellular fractions. The results indicated that PEMF could be developed as a potential therapy in the treatment of arthritis in humans.

Antiinflammatory Flavonoids from Artocarpus heterophyllus and Artocarpus communis

The antiinflammatory activities of the isolated flavonoids, including cycloartomunin (1), cyclomorusin (2), dihydrocycloartomunin (3), dihydroisocycloartomunin (4), cudraflavone A (5), cyclocommunin (6), and artomunoxanthone (7), and cycloheterohyllin (8), artonins A (9) and B (10), artocarpanone (11), artocarpanone A (12), and heteroflavanones A (13), B (14), and C (15) from Artocarpus communis and A. heterophyllus, were assessed in vitro by determining their inhibitory effects on the chemical mediators released from mast cells, neutrophils, and macrophages. Compound 4 significantly inhibited the release of beta-glucuronidase and histamine from rat peritoneal mast cells stimulated with P-methoxy-N-methylphenethylamine (compound 48/80). Compound 11 significantly inhibited the release of lysozyme from rat neutrophils stimulated with formyl-Met-Leu-Phe (fMLP). Compounds 8, 10, and 11 significantly inhibited superoxide anion formation in fMLP-stimulated rat neutrophils while compounds 2, 3, 5, and 6 evoked the stimulation of superoxide anion generation. Compound 11 exhibited significant inhibitory effect on NO production and iNOS protein expression in RAW 264.7 cells. The potent inhibitory effect of compound 11 on NO production in lipopolysaccharide (LPS)-activated macrophages, probably through the suppression of iNOS protein expression.

Human leukocyte elastase and cathepsin G are specific inhibitors of C5a‐dependent neutrophil enzyme release and chemotaxis

Circulating human neutrophils from patients with severe inflammatory disorders such as erysipelas and sepsis are specifically desensitized to complement factor C5a stimulation but not to stimulation with other stimuli like N-formyl-methionyl-leucyl-phenylalanine (FMLP), interleukin-8 (IL-8), leukotriene B4 (LTB4), or platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine). In this study, we raised the question whether factors released from polymorphonuclear leukocytes (PMNs) can specifically down-regulate C5a-dependent neutrophil functions. When neutrophils were preincubated with either neutrophil lysates or neutrophil degranulation supernatants, a complete inhibition of C5a-stimulated beta-glucuronidase release and chemotaxis could be observed, whereas FMLP-, IL-8-, LTB4- or PAF-dependent functions were not affected. Serine protease inhibitors like phenylmethylsulfonyl fluoride, antileukoprotease, or elafin abolished this effect. High-performance liquid chromatography of neutrophil degranulation supernatants revealed pronounced inhibition of C5a-dependent neutrophil functions in fractions exerting elastase or cathepsin G activity, but not in fractions exerting proteinase 3 activity. Using purified human leukocyte elastase (HLE), C5a responses like intracellular calcium influx, beta-glucuronidase release, and chemotaxis were also specifically inhibited. Our experiments show that the release of HLE or cathepsin G from neutrophils specifically down-regulates the responsiveness of neutrophils to C5a. Elastase and cathepsin G may therefore play an important role in the down-regulation of acute inflammation.

Nimbidin suppresses functions of macrophages and neutrophils: relevance to its antiinflammatory mechanisms.

Nimbidin is a mixture of tetranortriterpenes and is the major active principle of the seed oil of Azadirachta indica A. Juss (Meliaceae) possessing potent antiinflammatory and antiarthritic activities. The present study revealed that nimbidin significantly inhibited some of the functions of macrophages and neutrophils relevant to the inflammatory response following both in vivo and in vitro exposure. Oral administration of 5-25 mg/kg nimbidin to rats for 3 consecutive days significantly inhibited the migration of macrophages to their peritoneal cavities in response to inflammatory stimuli and also inhibited phagocytosis and phorbol-12-myristate-13-acetate (PMA) stimulated respiratory burst in these cells. In vitro exposure of rat peritoneal macrophages to nimbidin also inhibited phagocytosis and PMA stimulated respiratory burst in these cells. Nimbidin also inhibited nitric oxide (NO) and prostaglandin E2 (PGE2) production in lipopolysaccharide (LPS) stimulated macrophages following in vitro exposure, whereas interleukin 1 (IL-1) was only weakly inhibited. Probing the mechanism of NO inhibition revealed that nimbidin ameliorated the induction of inducible NO synthase (iNOS) without any inhibition in its catalytic activity. In addition, nimbidin also attenuated degranulation in neutrophils assessed in terms of release of beta-glucuronidase, myeloperoxidase and lysozyme. The results suggest that nimbidin suppresses the functions of macrophages and neutrophils relevant to inflammation. Thus nimbidin can be valuable in treating inflammation/inflammatory diseases.

Soluble guanylyl cyclase activator YC-1 inhibits human neutrophil functions through a cGMP-independent but cAMP-dependent pathway.

3-(5'-Hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1), a novel type of soluble guanylyl cyclase (sGC) activator, is useful in investigating the signaling of cGMP and may provide a new approach for treating cardiovascular diseases. Herein, YC-1 was demonstrated to inhibit the generation of superoxide anion (O2-) and the release of beta-glucuronidase release, to diminish the membrane-associated p47phox and to accelerate resequestration of cytosolic calcium in formyl-l-methionyl-l-leucyl-l-phenylalanine-activated human neutrophils. YC-1 not only directly promoted sGC activity and cGMP formation but also dramatically potentiated sodium nitroprusside-induced sGC activity and cGMP formation in human neutrophils. However, the synergistic increase in the amount of cGMP was inconsistent with its cellular response. Moreover, neither an sGC inhibitor nor protein kinase G inhibitors reversed the inhibitory effect of YC-1. Interestingly, YC-1 also increased the cAMP concentration and protein kinase (PK)A activity. The inhibitory effect of YC-1 was significantly enhanced by prostaglandin (PG)E1 and isoproterenol, and almost abolished by PKA inhibitors. These results show that cAMP, but not cGMP, mediates the YC-1-induced inhibition of human neutrophils. YC-1 increased the PGE1- and forskolin-induced but not 3-isobutyl-1-methylxanthine-produced cAMP formation, suggesting inhibition of phosphodiesterase. These findings thus reveal novel mechanism-mediated anti-inflammatory properties of YC-1 in human neutrophils, which can influence the progression of cardiovascular disease. cAMP, but not cGMP, plays an important role in the regulation of respiratory burst and degranulation in human neutrophils.

Terpenoids with a new skeleton and novel triterpenoids with anti-inflammatory effects from Garcinia subelliptica.

Three novel phloroglucinol derivatives, garcinielliptones F (1), H (3), and I (4), and two novel terpenoids, garcinielliptones G (2) and J (5), with a new skeleton have been isolated from the seeds of Garcinia subelliptica. Their structures, including relative configurations, were elucidated by spectroscopic methods and computer-generated molecular modeling. Compound 1 showed potent inhibitory effects on the release of beta-glucuronidase and lysozyme from rat neutrophils that had been stimulated with formyl-Met-Leu-Phe (fMLP)/cytochalasin B (CB). This effect was concentration-dependent with IC(50) values of 26.9+/-2.6 and 20.0+/-1.3 microM, respectively. Compound 1 also showed a potent concentration-dependent inhibitory effect on superoxide anion generation in rat neutrophils stimulated with fMLP/CB, with an IC(50) value of 17.0+/-0.9 microM. Compound 4 showed a potent inhibitory effect on NO production in culture media of N9 cells in response to lipopolysaccharide (LPS)/interferon-gamma (IFN-gamma) in a concentration-dependent manner with an IC(50) value of 7.4+/-0.2 microM.

Novel and anti-inflammatory constituents of Garcinia subelliptica.

Four novel phloroglucinol derivatives, garcinielliptones A (1), B (2), C (3), D (4), a novel triterpenoid, garcinielliptone E (5), and three known compounds were isolated from the seeds of Garcinia subelliptica. The structures, including relative configurations, were elucidated by means of spectroscopic data. Known compounds garsubellin A (6) and garcinielliptin oxide (7) showed potent inhibitory effects on the release of beta-glucuronidase, and beta-glucuronidase and histamine, respectively, from peritoneal mast cells stimulated with compound 48/80 in a concentration-dependent manner with IC(50) values of 15.6+/-2.5, and 18.2+/-3.6 and 20.0+/-2.7 microM, respectively. Compound 7 showed potent inhibitory effects on the release of beta-glucuronidase and lysozyme from neutrophils stimulated with formyl-Met-Leu-Phe(fMLP)/cytochalasin B (CB) in a concentration-dependent manner with IC(50) values of 15.7+/-3.0 and 23.9+/-3.2 microM, respectively. Compound 7 also showed potent inhibitory effect on superoxide formation from neutrophils stimulated with fMLP/CB also in a concentration-dependent manner with an IC(50) value of 17.9+/-1.5 microM.

Anti-inflammatory constituents and new pterocarpanoid of Crotalaria pallida.

One new pterocarpanoid, crotafuran E (1), and three known compounds were isolated from the bark of Crotalaria pallida. The structure of 1 was determined by spectral methods. Two pterocarpanoids, crotafurans A (2) and B (3), previously isolated from this plant, showed significant concentration-dependent inhibitory effects on the NO production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage-like cells, with IC(50) values of 23.0 +/- 1.0 and 19.0 +/- 0.2 microM, respectively. Compound 3 also inhibited the LPS/interferon-gamma (IFN-gamma)-stimulated NO production in N9 microglial cells with an IC(50) value of 9.4 +/- 0.9 microM. Moreover, compound 2 produced a concentration-dependent inhibition of the release of beta-glucuronidase and lysozyme from rat neutrophils in response to formyl-Met-Leu-Phe/cytochalasin B (fMLP/CB) with IC(50) values of 7.8 +/- 1.4 and 9.5 +/- 2.1 microM, respectively.

The effect of selenium on immune functions of J774.1 cells.

The J774.1 macrophage cell line was used as a tool to investigate the influence of selenium on macrophage function. In vitro selenium supplementation enhanced phagocytosis, degranulation by the release of beta-glucuronidase after N-formyl-methionyl-leucyl-phenylalanine (FMLP) or cytochalasin B, and the production of superoxide anion after phorbol myristate acetate stimulation of these cells, while the release of nitric oxide was not affected by the selenium status. Selenium supplementation enhanced significantly (p < 0.05) the release of tumor necrosis factor (5-fold), interleukin-1 (3-fold) and interleukin-6 (2.5-fold) after 10 microg/ml lipopolysaccharide stimulation compared to selenium-deficient cells.