Supinya Tewtrakul

Effects of compounds from Garcinia mangostana on inflammatory mediators in RAW264.7 macrophage cells

ETHNOPHARMACOLOGICAL RELEVANCE The fruit hull of Garcinia mangostana Linn. has been used in Thai traditional medicine for treatment of abscess and skin infection. AIM OF THE STUDY The mangosteen fruit hull and its compounds were carried out to investigate for anti-inflammatory activity. MATERIAL AND METHODS The extract of Garcinia mangostana together with alpha- and gamma-mangostins were tested for anti-inflammatory effect against lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-alpha) and interleukin-4 (IL-4) releases as well as their mechanisms in transcriptional levels using RAW264.7 macrophage cells. RESULTS Mangosteen extract possessed potent NO inhibitory effect with an IC50 value of 1.0 microg/ml. The isolated compounds from the extract including alpha-mangostin and gamma-mangostin, possessed marked inhibitory effect against NO release with IC50 values of 3.1 and 6.0 microM, respectively. The extract exhibited potent inhibitory effect on PGE2 release (IC50=6.0 microg/ml), whereas those of alpha- and gamma-mangostins were 13.9 and 13.5 microM, respectively. However, mangostins possessed only moderate effects towards TNF-alpha and IL-4 releases with IC50 values ranging from 31.8 to 64.8 microM. Both extract and alpha-mangostin suppressed transcription of gene encoding inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in dose-dependent manners, whereas gamma-mangostin had only an inhibitory effect on transcription of iNOS. CONCLUSION The present study may support the Thai traditional use of Garcinia mangostana fruit hull for treatment of inflammatory-related diseases through the inhibition of NO and PGE2 releases, but moderate effect through TNF-alpha and IL-4.

Suppressive effects of methoxyflavonoids isolated from Kaempferia parviflora on inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells

ETHNOPHARMACOLOGICAL RELEVANCE The rhizomes of Kaempferia parviflora Wall. ex Baker have been traditionally used in Thailand to treat abscesses, gout, and peptic ulcers. AIM Previously, we reported that the chloroform fraction of a Kaempferia parviflora extract had an inhibitory effect on rat paw-edema. In the present study, we isolated the constituents of this fraction and investigated the anti-inflammatory mechanism against nitric oxide (NO) production, tumor necrosis factor-α (TNF-α) and the expression of inducible nitric oxide synthase (iNOS) as well as phosphorylated extracellular signal-regulated kinase (p-ERK), and phosphorylated c-Jun N-terminal kinase (p-JNK). In addition, effects of trimethylapigenin (4) on the enzyme activities of protein kinases possibly leading to iNOS expression were examined to clarify the targets. MATERIALS AND METHODS The chloroform fraction was isolated using silica gel column chromatography and HPLC. Isolated compounds were tested against NO and TNF-α using RAW264.7 cells. Cytotoxicity and iNOS, p-ERK and p-JNK expression were also examined. RESULTS Three active components, 5,7-dimethoxyflavone (2), trimethylapigenin (4), and tetramethylluteolin (5), markedly inhibited the production of NO in lipopolysaccharide (LPS)-activated RAW264.7 cells. Compounds 2, 4, and 5 moderately inhibited production of TNF-α. Compounds 2, 4, and 5 strongly inhibited expression of iNOS mRNA and iNOS protein in a dose-dependent manner, but did not inhibit p-ERK or p-JNK protein expression. The most active compound, 4, did not inhibit the enzyme activity of inhibitor of κB kinases or mitogen-activated protein kinases, but inhibited that of spleen tyrosine kinase (SYK). CONCLUSION The mechanism responsible for the anti-inflammatory activity of methoxyflavonoids from the chloroform fraction of the rhizomes of Kaempferia parviflora is mainly the inhibition of iNOS expression, and the inhibition of SYK by 4 may be involved in the suppression of LPS-induced signaling in macrophages.

Potential anti-inflammatory diterpenoids from the roots of Caesalpinia mimosoides Lamk.

Anti-inflammatory assay-guided separation of extracts from the roots of Caesalpinia mimosoides Lamk. led to isolation of seven compounds: four diterpenes (1-4), a dimer (9), and two dibenzo[b,d]furans (10, 11) together with eleven known compounds. Their structures were elucidated by 1D- and 2D-NMR techniques as well as UV, IR, mass spectral data and comparison with literature values. The anti-inflammatory activities of all compounds were evaluated for inhibitory activities against lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW264.7 cell lines. Compounds 4, 6, 8, and 12-14 were also tested for the inhibitory effect on LPS-induced tumor necrosis factor-alpha (TNF-alpha) release in RAW264.7 cells. The results indicated that 4 possessed potent inhibitory activity for both tests with IC(50) values of 3.0 and 6.5 microM, respectively.

Anti‐allergic activity of principles from the roots and heartwood of caesalpinia sappan on antigen‐induced β‐hexosaminidase release

The dichloromethane extract of the roots and heartwood of Caesalpinia sappan exhibited potent inhibitory activity against β‐hexosaminidase release as marker of degranulation in rat basophilic leukemic (RBL‐2H3) cells, with inhibition of 98.7% and 87.5% at concentration of 100 µg/ml, respectively. These extracts were further separated by chromatographic techniques to give two chalcones and seven homoisoflavones. Among the compounds tested, sappanchalcone (2) possessed the most potent effect against allergic reaction in RBL‐2H3 cells with an inhibitory concentration (IC50) value of 7.6 µM, followed by 3‐deoxysappanchalcone (1, IC50 = 15.3 µM), whereas other compounds showed moderate and mild effects. The results suggested the following structural requirements of chalcones (1 and 2) and homoisoflavones (3‐9) for anti‐allergic activity: (i) chalcone exhibited higher activity than homoisoflavone (ii) vicinal hydroxylation at B‐ring of chalcone conferred higher activity than one hydroxylation; and (iii) for homoisoflavone, the hydroxyl groups at C‐3 and C‐4 positions decreased the activity. This is the first report of C. sappan for anti‐allergic activity. Copyright

Plumbagin production by root cultures of Plumbago rosea

Root cultures of Plumbago rosea Linn. were established from young leaf explants on solid Gamborgs B5 (B5) medium supplemented with the combination of α-naphthalene acetic acid (NAA) and kinetin in the concentration ranges of 0.5-2.0 mg/l and 0.1-0.5 mg/l, respectively. The production of plumbagin, determined by TLC-densitometry was higher [0.016 ± 0.0030% dry weight (DW)] in cultured roots obtained from B5 medium supplemented with 1.0 mg/l NAA and 0.1 mg/l kinetin. Plant selection increased the plumbagin production to 0.129 ± 0.0139% DW, while variation of sucrose and nitrogen (as (NH 4 ) 2 SO 4 ) concentration in B5 media slightly increase the plumbagin synthesis to 0.023 ± 0.0017 and 0.020 ± 0.0015% DW, respectively.

Anti-allergic principles of Rhinacanthus nasutus leaves

Three naphthoquinone derivatives, rhinacanthin-C (1), -D (2) and -N (3) were isolated from the extract of Rhinacanthus nasutus Kurz leaves and were tested for anti-allergic effect. The result indicated that all three compounds possessed very potent anti-allergic activity against antigen-induced beta-hexosaminidase release as a marker of degranulation in RBL-2H3 cells with IC(50) values of 6.9, 8.9 and 6.4 microM, respectively. In addition, the effects of rhinacanthin-C, -D and -N on antigen-induced release of TNF-alpha and IL-4 were also examined. It was found that rhinacanthin-C showed the most potent on antigen-induced TNF-alpha release with an IC(50) value of 0.7 microM, followed by rhinacanthin-D (IC(50)=3.8 microM) and rhinacanthin-N (IC(50)=10.3 microM), whereas those for IL-4 were rhinacanthin-D (IC(50)=5.4 microM), rhinacanthin-C (IC(50)=7.0 microM) and rhinacanthin-N (IC(50)=12.0 microM), respectively. The mechanisms in the late phase reaction of rhinacanthin-C, -D and -N were found to inhibit TNF-alpha and IL-4 gene expression in antigen-induced TNF-alpha and IL-4 releases on from RBL-2H3 cells as dose-dependent manners. The structure-activity trends of rhinacanthin-C,-D and-N on the inhibition of TNF-alpha release are as follow; substitution with octadienoic acid (rhinacanthin-C) conferred much higher activity than that of benzodioxo carboxylic acid ester (rhinacanthin-D) as well as naphthalene carboxylic acid ester (rhinacanthin-N). For the inhibition of IL-4 release, the substitution with octadienoic acid (rhinacanthin-C) and benzodioxo carboxylic acid ester (rhinacanthin-D) possessed the effect two-fold higher than that of naphthalene carboxylic acid ester (rhinacanthin-N). As regards active constituents for anti-allergic activity of R. nasutus, rhinacanthin-C, -D and -N are responsible for anti-allergic effect of this plant on both the early phase and late phase reactions. The finding may support the traditional use of R. nasutus leaves for treatment of allergy and allergy-related diseases.

Diterpenoids and triterpenoids with potential anti-inflammatory activity from the leaves of Aglaia odorata.

Chemical investigation of the leaves of the oriental medicinal plant Aglaia odorata resulted in the isolation of five compounds: two dolabellane diterpenoids, two dammarane triterpenoids and a protostane triterpenoid, along with twenty known compounds. Their structures were elucidated on the basis of extensive spectroscopic analysis and by comparison of their NMR spectroscopic data with those reported in the literature. The anti-inflammatory activities of all compounds were evaluated as inhibitory activities against lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW264.7 cell lines. Eleven compounds possessed potent nitric oxide inhibitory activity with IC(50) values ranging from 2.1 to 14.2 μM, these being better than that of the positive control, indomethacin (IC(50)=14.5 μM). In addition, three compounds exhibited significant activity against PGE(2) release with IC(50) values of 2.6, 16.1 and 23.0 μM.

Antiinflammatory constituents from Eclipta prostrata using RAW264.7 macrophage cells.

The whole plant extract of Eclipta prostrata and its isolated compounds were tested for their antiinflammatory effects against lipopolysaccharide (LPS)‐induced nitric oxide (NO), prostaglandin E2 (PGE2) and tumor necrosis factor‐alpha (TNF‐α) release in RAW264.7 cells, as well as for the antiinflammatory mechanism of the active compound on mRNA expression. Among the isolated compounds, orobol (5) exhibited the highest activity against NO release with an IC50 value of 4.6 μm, followed by compounds 1, 2 and 4 with IC50 values of 12.7, 14.9 and 19.1 μm, respectively. The IC50 value of compound 5 against PGE2 release was found to be 49.6 μm, whereas it was inactive towards TNF‐α (IC50 > 100 μm). The mechanism of orobol (5) was found to down‐regulate iNOS and COX‐2 mRNA expression in a concentration‐dependent manner. The present study may support the traditional use of Eclipta prostrata for the treatment of inflammatory‐related diseases. Copyright

Fruit Oil Composition of Piper chaba Hunt., P. longum L. and P. nigrum L.

Abstract Essential oil composition of the fruit oils of Piper chaba, P. longum and P. nigrum were investigated by GC and GC/MS. Unlike P. nigrum oil, P. chaba and P. longum oils were found to contain few monoterpene hydrocarbons, a moderate content of sesquiterpenes and high content of aliphatic hydrocarbons. The content of caryophyllene oxide (7.4%) was the highest in P. chaba and that of β-caryophyllene was found to be 39.7%, 10.2% and 5.2% in, P. nigrum, P. longum and P. chaba, respectively.

Potential anti-allergic acridone alkaloids from the roots of Atalantia monophylla.

Acridone alkaloids, cycloatalaphylline-A (1), N-methylcyclo-atalaphylline-A (2) and N-methylbuxifoliadine-E (3), were isolated from the dichloromethane and acetone extracts of the root of Atalantia monophylla along with eight known acridone alkaloids: buxifoliadine-A (4), buxifoliadine-E (5), N-methylatalaphylline (6), atalaphylline (7), citrusinine-I (8), N-methylataphyllinine (9), yukocitrine (10) and junosine (11) and two known coumarins: auraptene (12) and 7-O-geranylscopoletin (13). Their structures were elucidated on the basis of spectroscopic analyses. Compounds 2, 5 and 8 possessed appreciable anti-allergic activity in RBL-2H3 cells model with IC(50) values of 40.1, 6.1 and 18.7 microM, respectively.