LPS-induced NO Production In RAW264 Research Articles

Chemical constituents of Abies delavayi

Systematic phytochemical investigations on Abies delavayi afforded 110 compounds, including 49 terpenoids, 13 lignans, 20 flavonoids, three coumarins, and 25 other chemical constituents. By detailed analysis of one- and two-dimensional NMR spectroscopic and high-resolution mass spectrometric data, 10 previously unreported compounds were identified: they comprised three sesquiterpenoids, two diterpenoids, one triterpenoid, one monoterpenoid, one flavonoid, and two phenols. These 10 compounds and some previously known ones were subjected to two cytotoxic bioassays against three human tumor cell lines and NO production inhibition on RAW264.7 macrophages, respectively. (25R)-24,25-Dihydroabieslactone had the strongest cytotoxic activity against Colo-205 cells with an IC50 value of 19.0±3.7μg/mL. (+)-T-cadinol, 8,11,13-abietatrien-15-ol-18-yl acetate, 18-acetoxy-13-epi-manool, imperatorin, bergapten, and 5,7-O-dimethyl poriol exhibited weak inhibitory activity against LPS-induced NO production in RAW264.7 macrophages with IC50 values of approximately 50μg/mL.

A New Noreudesmane-type Sesquiterpenoid from Alpinia oxyphylla

isolation, structure determination, and inhibitory effects of the new noreudesmane-type sesquiterpenoid 4-methoxy-oxyphyllenone A (I) and fifteen known compounds on NO production in LPS-induced RAW 264.7 cells

Chemical constituents from the aerial parts of Psammosilene tunicoides

Investigation of the 80% ethanol extract of the aerial parts of Psammosilene tunicoide afforded two carboline alkaloids (1–2), a γ-pyrone derivative (3), two p-hydroxy cinamic acid derivatives (4–5), two glycosides (6–7) and six known compounds (8–13). Their structures were established on the basis of spectroscopic analysis (1D-, 2D-NMR, ESI-MS, and HR-ESI-MS). All these compounds were evaluated for cytotoxicity against SMML-7721, A549, HCT116, and MCF-7 cell lines, and for inhibitory activity against NO production in LPS-induced RAW264.7 macrophages. However, only compound 2 had weak cytotoxic activity against the HCT116 cell line at 20μM, while the others are inactive against four human tumor cell lines and only show weak inhibition activity against NO production at 50μM.

Three new sesquiterpene lactones from Inula britannica.

One new 1,10-secoeudesmanolide (1), two eudesmanolides (2 and 3), together with nine known compounds (4-12) were isolated from the aerial parts of Inula britannica. The structures of the new compounds were elucidated by detailed spectroscopic analysis, including HRESIMS and 2D-NMR spectroscopic method. In addition, compounds 1-4 were tested for their inhibitory effects against LPS-induced NO production in RAW264.7 macrophages.

New triterpenic acids from Uncaria rhynchophylla: Chemistry, NO-inhibitory activity, and tandem mass spectrometric analysis

Five new oleanane and ursane type triterpenes, namely uncarinic acids F–J (1–5), together with six known triterpenic acids (6–11) were isolated from the stems and hooks of Uncaria rhynchophylla. Structure elucidation of 1–5 was based on the integrated analyses of high-resolution MS data, 1D (1H NMR, 13C NMR, DEPT) and 2D (HSQC, HMBC, ROESY) NMR spectra. Compounds 4, 10, and 11 exhibited weak inhibitory effects on LPS-induced NO production in RAW264.7 cells (with IC50 1.48, 7.01, and 1.89μM, respectively) with dexamethasone (IC50 0.04μM) and quercetin (IC50 0.86μM) as the positive controls. 19-OH substituted oleanane triterpenic acids (1, 2, 5, 8) were prone to eliminate CH2O3, whereas those ursane-type encompassing 19-OH (3, 6, 7, 9, 4) were featured by preferred cleavage of H2O while performing the negative collision-induced MS/MS fragmentation on an LTQ/Orbitrap mass spectrometer.

Lignans and neolignans from the stems of Vibrunum erosum and their neuroprotective and anti-inflammatory activity

A new lignan, (7'S,8S,8'S)-3,5'-dimethoxy-3',4,9'-trihydroxy-7',9-epoxy-8,8'-lignan, named vibruresinol (1), was isolated from the stems of Viburnum erosum by silica gel, ODS, and Sephadex LH-20 column chromatography. In addition, four other known lignans, (7'R,8S,8'S)-3,5'-dimethoxy-3',4,8',9'-tetrahydroxy-7',9-epoxy-8,8'-lignan (2), (+)-syringaresinol (3), (+)-pinoresinol (4), and (+)-pinoresinol-4-O-β-D-glucopyranoside (5), and five known neolignans, herpetol (6), vibsanol (7), (-)-dehydrodiconiferyl alcohol (8), icariside E4 (9), and dihydrodehydrodiconiferyl alcohol (10), were isolated in the same manner. The chemical structures of the compounds were determined based on spectroscopic data including NMR, MS, and IR. All of the compounds described above were isolated from V. erosum for the first time. The isolated compounds 3, 4, and 6 were evaluated for neuroprotective activity on glutamate-induced cell death in HT22 cells and had EC50 values of 6.33±1.22, 6.96±0.65, and 9.15±0.36μM, respectively. Likewise, the same compounds had inhibitory activity on NO production in LPS-induced RAW 264.7 cells with IC50 values of 8.30±1.56, 7.89±1.22, and 9.32±0.36μM, respectively.

New phenylpropanoid and other compounds from Illicium lanceolatum with inhibitory activities against LPS-induced NO production in RAW 264.7 macrophages

Illicium lanceolatum is a traditional Chinese medicine (TCM) for treating inflammatory diseases. Anti-inflammatory activities of I. lanceolatum stems and leaves were tested using ear edema models induced by dimethyl benzene in mice. Bioassay-guided fractionation of the ethanol extract of I. lanceolatum leaves and stems revealed that the ethyl acetate fraction exhibited inhibitory potency to dimethyl benzene-induced edema in the mouse ear. Phytochemical investigation on the active fraction led to the isolation of a new phenylpropanoid (1), together with fifteen known compounds. This is the first report of the isolation of 2–16 from I. lanceolatum. Of these compounds, compounds 1, 2 and 3 showed inhibitory activity on LPS-stimulated NO production in RAW 264.7 macrophages with IC50 values of 27.58, 26.59 and 34.35μg/mL, respectively. I. lanceolatum stems and leaves can be exploited to alleviate inflammatory diseases, which makes the rare medicinal plant resources sustainable.

Inhibitory effects of new varieties of bitter melon on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells

Bitter melon is widely used as a food and a traditional herbal medicine for the treatment of diabetes; it also possesses antioxidant, anti-tumour, anti-bacterial, anti-obesity, and anti-hypertension activities. The aim of this work was to study the in vitro anti-inflammatory effects of different solvent extracts of the seeds and fruits of new varieties of bitter melon. Of the 30 bitter melon extracts tested, the ethyl acetate extract of the fruit of Momordica charantia MDS72 (EAEF-MCMDS72) exhibited the strongest anti-inflammatory activity. EAEF-MCMDS72 significantly inhibited the production of NO, IL-6, PGE2, TNF-α, and MCP-1 in lipopolysaccharide (LPS)-treated RAW 264.7 cells. The expression levels of the iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells were inhibited by EAEF-MCMDS72. Real-time RT-PCR analysis indicated that the mRNA expression levels of iNOS, COX-2, IL-6, and TNF-α were suppressed by EAEF-MCMDS72. Moreover, four known triterpene compounds including (23E)-cucurbita-5,23,25-triene-3β,7β-diol (1), (23E)-25-methoxycucurbit-23-ene-3β,7β-diol (2), (23E)-5β,19-epoxycucurbita-6,23-diene-3β,25-diol (3), and 3,7-dioxo-23,24,25,26,27-pentanorcucurbit-5-en-22-oic acid (4) were isolated from the fruit of M. charantia MDS72. Compounds 2, 3, 4 suppressed the LPS-induced NO production in RAW 264.7 cells. Furthermore, the present study may contribute to our understanding of the anti-inflammatory effects of EAEF-MCMDS72 and its triterpene compounds.

Neolignans from the Fruits of Magnolia obovata and Their Inhibition Effect on NO Production in LPS-Induced RAW 264.7 Cells

Three new neolignans, named 9-methoxyobovatol (6), magnobovatol (7), and 2-hydroxyobovaaldehyde (9), along with six known ones, magnolol (1), honokiol (2), isomagnolol (3), obovatol (4), obovatal (5), and obovaaldehyde (8), were isolated from the fruits of Magnolia obovata using silica gel and ODS column chromatography. From the results of spectroscopic data including EIMS, IR, 1H- and 13C-NMR, DEPT, and 2D-NMR (gCOSY, gHSQC, gHMBC), the chemical structures were determined. All isolated compounds were evaluated for inhibition activity on nitric oxide production in LPS-induced RAW 264.7 cells, and compounds 1-4, 6, 7, and 9 showed significant activity with IC50 values of 15.8 ± 0.3, 3.3 ± 1.2, 14.1 ± 0.9, 6.2 ± 1.2, 14.8 ± 2.3, 14.2 ± 1.2, and 14.8 ± 3.2 µM, respectively, without any visible toxic effect.

Inhibitory effect of resveratrol dimerized derivatives on nitric oxide production in lipopolysaccharide-induced RAW 264.7 cells

Four types of resveratrol dimerized analogues were synthesized and evaluated in vitro on LPS-induced NO production in RAW 264.7 cells. The results showed that several compounds, especially those containing 1,2-diphenyl-2,3-dihydro-1H-indene core (type I), exhibited good inhibitory activities. Among 25 analogues, 12b showed a significant inhibitory activity (49% NO production at 10μM, IC50=3.38μM). Further study revealed that compound 12b could suppress LPS-induced iNOS expression, NO production, and IL-1β release in a concentration-dependently manner. The mechanism of action (MOA) involved for its anti-inflammatory responses was through signaling pathways of p38 MAPK and JNK1/2, but not ERK1/2.

Chemical constituents from Inula wissmanniana and their anti-inflammatory activities

Two new inositol derivatives, 4-acetate-1,2,3,5-tetrakis(2-methyl-2-butenoate) inositol (1) and 3-acetate-1,2,4,5-tetrakis(2-methyl-2-butenoate) inositol (2), one new flavonolignan 23-o-acetylsilychristin A (3), and one new thymol derivative 3-hydroxy-4-(2-hydroxy-1-methylethyl)benzoic acid (4), along with 13 known compounds, including ten flavonolignans (5-14) and three thymol derivatives (15-17) were isolated from the aerial part of Inula wissmanniana. The structures of new compounds were elucidated by detailed spectroscopic analysis, including HRESIMS and 2D-NMR spectroscopic data. All the compounds were isolated from this plant for the first time. The anti-inflammatory activities of the compounds were tested against LPS-induced NO production in RAW 264.7 macrophages.

Five new sesquiterpene lactones from Inula hupehensis

Four new pseudoguaianolides (1-4), one new guaianolide (5), together with ten known compounds (6-15) were isolated from the aerial parts of Inula hupehensis. Their structures were elucidated mainly on the basis of 1D and 2D spectroscopic methods and circular dichroism analysis. In addition, compounds 1-10 and 13 were tested for their inhibitory effects against LPS-induced NO production in RAW264.7 macrophages. Compounds 2, 6, 8 and 9 exhibited significant inhibitory activities with IC50 values in the range of 0.6-6.6 μM.

Microbial transformation of acetyl-11-keto-β-boswellic acid and their inhibitory activity on LPS-induced NO production

The capabilities of 20 strains of fungi to transform acetyl-11-keto-β-boswellic (AKBA) were screened. And biotransformation of AKBA by Cunninghamella blakesleana AS 3.970 afforded five metabolites (1–5), while two metabolites (6, 7) were isolated from biotransformation of Cunninghamella elegans AS 3.1207. The chemical structures of these metabolites were identified by spectral methods including 2D NMR and their structures were elucidated as 7β-hydroxy-3-acety-11-keto-β-boswellic acid (1), 21β-dihydroxy-3-acety-11-keto-β-boswellic acid (2), 7β,22α-dihydroxy-3-acety-11-keto-β-boswellic acid (3), 7β,16α-dihydroxy-3-acety-11-keto-β-boswellic acid (4), 7β,15α-dihydroxy-3-acety-11-keto-β-boswellic acid (5); 7β,15α,21β-trihydroxy-3-acety-11-keto-β-boswellic acid (6) and 15α,21β-dihydroxy-3-acety-11-keto-β-boswellic acid (7). All these products are previously unknown. Their primary structure–activity relationships (SAR) of inhibition activity on LPS-induced NO production in RAW 264.7 macrophage cells were evaluated.

Isolation of a new meroterpene and inhibitors of nitric oxide production from Psoralea corylifolia fruits guided by TLC bioautography

A new meroterpene, psoracorylifol F (1), was isolated from Psoralea corylifolia fruits, along with two known meroterpenes (2 and 3), guided by TLC bioautography against O2− radicals. The structure of 1 was elucidated by means of NMR, HRESIMS, and X-ray crystallographic analysis. All the three metroterpenes possessed potential inhibitory activity against LPS-induced NO production in RAW 264.7 cells with IC50 values ranging from 7.71 to 27.63μM.

Prenylated phloroglucinol derivatives from Hypericum sampsonii

Six new acylphloroglucinol derivatives, sampsonols A-F (1–6), were isolated from the petroleum ether extract of the aerial parts of Hypericum sampsonii. The structures and relative configurations of sampsonols A-F were elucidated by extensive spectroscopic analyses. All these compounds were tested for their in vitro cytotoxic and anti-inflammatory activities. Sampsonols A and B (1 and 2) showed significant cytotoxicity against four human tumor cell lines with IC50 values in the range of 13–28μM, whereas sampsonols C and F (3 and 6) showed potent inhibitory activities against LPS-induced NO production in RAW 264.7 macrophages with IC50 values of 27.3 and 29.3μM, respectively.

Microbial transformation of deoxyandrographolide and their inhibitory activity on LPS-induced NO production in RAW 264.7 macrophages

A series of analogues of deoxyandrographolide (1) transformed by Cunninghamella blakesleana AS 3.2004 were isolated and identified by spectral methods including 2D NMR. Among them, 3-oxo-17,19-dihydroxy-7,13-ent-labdadien-15,16-olide (9), 3-oxo-19-hydroxy-1,13-ent-labdadien-15,16-olide (16), 3-oxo-1β-hydroxy-14-deoxy-andrographolide (17) and 3-oxo-2β-hydroxy-14-deoxyandrographolide (18) are new compounds. And their structure–activity relationships (SAR) of inhibitory activity on LPS-induced NO production in RAW 264.7 macrophage cells were also discussed.

Antioxidant, Anti-inflammatory, and Antiproliferative Activities of Taxillus sutchuenensis

Inflammation is related to several chronic diseases, including cancer and atherosclerosis. Taxillus sutchuenensis (Lecomte) Danser is a special folk medicinal plant in Taiwan. The aim of this study was to evaluate the antioxidant, anti-inflammatory, and antiproliferative activities of the aqueous-thanol extract from T. sutchuenensis (AETS) and its fractions. TEAC, DPPH radicals, total phenolic compounds, total flavonoid content, inhibition of NO production in LPS-induced RAW264.7 cells, and inhibition of cancer cell proliferation were tested. Among all fractions, the ethyl-acetate (EA) fraction showed the highest TEAC and DPPH radical scavenging activities. The EA fraction also had the highest polyphenol and flavonoid content. The EA fractions also decreased LPS-induced NO production and the expression of iNOS and COX-2 in RAW264.7 cells. The antiproliferative activities of the aqueous/ethanol extract and fractions were studied in vitro using A549 cells, and the results were consistent with their antioxidant capacities. EA fractions had the highest antiproliferative activity with an IC(50) of 454.38 ± 1.48 μg/ml. Quercetin also had antioxidant, anti-inflammatory, and antiproliferative activities. Quercetin might be an important bioactive compound in T. sutchuenensis. The experimental data indicated that T. sutchuenensis is a potent antioxidant medicinal plant, and such efficacy may be mainly attributed to its polyphenolic compounds.

Alkaloids from Chelidonium majus and their inhibitory effects on LPS-induced NO production in RAW264.7 cells

A new alkaloid, methyl 2'-(7,8-dihydrosanguinarine-8-yl)acetate (1), together with six known alkaloids, stylopine (2), protopine (3), norchelidonine (4), chelidonine (5), berberine (6), and 8-hydroxydihydrosanguinarine (7), were isolated from Chelidonium majus. Their chemical structures were primarily established using 1D and 2D NMR techniques and mass spectrometry. The anti-inflammatory activity of the isolates was examined for their inhibitory effects on LPS-induced NO production in macrophage RAW264.7 cells. Among them, compounds 5 and 7 showed strong inhibitory activities toward the LPS-induced NO production in macrophage RAW264.7 cells with IC(50) values of 7.3 and 4.5 μM, respectively. In addition, compounds 5 and 7 inhibited the inductions of COX-2 and iNOS mRNA in dose-dependent manners, indicating that these compounds attenuated the syntheses of these transcripts at the transcriptional level.

Cyclopenicillone, a unique cyclopentenone from the cultures of Penicillium decumbens

Cyclopenicillone (1), possessing a unique 2,5-dimethylcyclopent-2-enone carbon skeleton, has been isolated from the cultures broth of the fungus Penicillium decumbens. The structure and absolute configuration of 1 were elucidated using a combination of NMR spectroscopy, CD data and computational approaches. Cyclopenicillone (1) demonstrated a dose-dependent (10-100 μM) inhibition against LPS-induced NO production in RAW264.7 macrophages.

Antioxidant, Anti-inflammatory and Antiproliferative Activities of Kalanchoe gracilis (L.) DC Stem

Oxidative stress and inflammation are related to several chronic diseases including cancer and atherosclerosis. Kalanchoe gracilis (L.) DC is a special folk medicinal plant in Taiwan. The aim of this study was to evaluate the antioxidant, anti-inflammatory and antiproliferative activities of the methanolic extract and fractions of the stem of K. gracilis. TEAC, total phenolic compound content, total flavonoid content, DPPH radical scavenging activity, reducing power, inhibition of NO production in LPS-induced RAW264.7 cells, and inhibition of cancer cell proliferation were analyzed. Among all fractions, the chloroform fraction showed the highest TEAC and DPPH radical scavenging activities. The chloroform fraction also had the highest content of polyphenols and flavonoids. Chloroform fractions also decreased LPS-induced NO production and expressions of iNOS and COX-2 in RAW264.7 cells. The antiproliferative activities of the methanolic extract and fractions were studied in vitro using HepG2 cells, and the results were consistent with their antioxidant capacities. Chloroform fractions had the highest antiproliferative activity with an IC(50) of 136.85 ± 2.32 μg/ml. Eupafolin also had good pharmacological activity in the antioxidant, anti-inflammation and antiproliferative. Eupafolin might be an important bioactive compound in the stem of K. gracilis. The above experimental data indicated that the stem of K. gracilis is a potent antioxidant medicinal plant, and such efficacy may be mainly attributed to its polyphenolic compounds.