Anti-inflammatory in vitro activities of eleven selected caffeic acid derivatives based on a combination of pro-/anti-inflammatory cytokine secretions and principal component analysis - A comprehensive evaluation.

Eleven compounds including caffeic acid (CA), 4 kinds of caffeoylquinic acid (CQA) and 6 kinds of dicaffeoylquinic acid (DCQA), were selected to evaluate the anti-inflammatory effectiveness using mouse primary peritoneal macrophages in the absence or presence of lipopolysaccharide (LPS). The optimal non-cytotoxic doses of each individual compound were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Pro-inflammatory (TNF-α, IL-1ß, IL-6) and anti-inflammatory (IL-10) cytokines secreted by treated macrophages were analyzed using the enzyme-linked immunosorbent assay. Cytokine secretion profiles of each individual test sample at optimal non-cytotoxic doses were further analyzed using Principal Component Analysis (PCA). The results showed that CA and all selected CQAs exhibited lower cytotoxicity (IC50: >50 µmol/l). Both CA and 5-CQA were found to have the most significant contributions for inhibiting pro-inflammatory cytokines, but increasing anti-inflammatory cytokine secretions, evidencing that CA at 10 µmol/l and 5-CQA at 25 µmol/l can be qualified as potent anti-inflammatory agents for treating inflammation-related diseases.

Anti-inflammatory effects of a naturally lacto-fermented cucumber product on RAW 264.7 macrophages in association with increased functional ingredients.

A naturally lacto-fermented cucumber product was developed for use as anti-inflammatory functional foods. To explore the anti-inflammatory characteristics, water (CWE) and ethanol extracts (CEE) from this product were selected to assess their anti-inflammatory potential on RAW 264.7 macrophages in the absence or presence of lipopolysaccharide (LPS), using four different inflammatory models. Changes in pro- (IL-1ß, IL-6 and TNF-α) and anti-inflammatory (IL-10) cytokine secretions by treated macrophages were measured using ELISA. The results showed that both CWE and CEE had strong potential to inhibit LPS-stimulated inflammation in macrophages in a repair manner. CWE had a better effect than CEE. The total phenolic, flavonoid and saponin contents in CEE were significantly (P < 0.05) correlated with IL-10 (r = 0.384, P = 0.036*) and TNF-α (r = 0.371, P = 0.043*) levels, but slightly correlated with TNF-α/IL-10 secretion ratios (r = -0.184, P = 0.359) by treated RAW 264.7 cells, respectively.

Culture condition optimization of naturally lacto-fermented cucumbers based on changes in detrimental and functional ingredients.

A two-step trial was used to optimize the culture condition of naturally lacto-fermented cucumbers. In the first trial, changes in pH values and total biogenic amines were measured to optimize the pickling juice formula. A 15% crystal sugar solution with low-salt brine at 4 °C was proved to be the best formula. In the second trial, pH values, organic acids, total phenolics, flavonoids, saponins and free amino acids, as well as biogenic amines and nitrites under the optimal pickling formula were measured. The optimal fermentation day was suggested at around 8 days. During the cucumber's fermentation process, the pH value was quickly lowered to <4.6. Meanwhile, the functional ingredients increased significantly. In contrast, total biogenic amines and nitrites did not exceed the risk limit, evidencing the safety and functional characteristics for the naturally lacto-fermented cucumbers. The two-step trial has evidenced the possibility to develop desirable lacto-fermented cucumbers.

Menthone supplementation protects from allergic inflammation in the lungs of asthmatic mice.

To screen potent terpenoid compounds against allergic inflammation in vitro and in vivo, five terpenoid compounds including menthone, farnesol, oridonin, ß-escin and lupeol, were first selected to compare their anti-allergic inflammation potential using mouse lung mast cells in vitro. Among five selected terpenoid compounds, just menthone treatment decreased TNF-α/IL-10 secretion ratios in lipopolysaccharide -stimulated mast cells in vitro. As a result, menthone was further chosen to treat ovalbumin (OVA)-sensitized and challenged BALB/c mice by gavage for 5 weeks. There were six groups including dietary control (DC group, 0 mg menthone/kg b.w./day), 8 (ML group), 40 (MM group) as well as 200 mg menthone/kg b.w./day (MH group) by gavage, positive control (PC group, 3 mg dexamethasone/kg b.w. by gavage before OVA challenge) and non-treatment control (NTC group, normal mice without treatment) in the experiment. Changes of inflammatory mediators, cell distribution, Th1/Th2 and pro-/anti-inflammatory cytokines secretion as well as relative gene expression amounts of six receptors related to allergic inflammation in the lungs and airways were measured. The results showed that middle menthone supplementation (40 mg menthone/kg b.w./day) in vivo decreased protein and eotaxin, but increased Th1 cytokine levels in the bronchoalveolar lavage fluid. Menthone supplementation inhibited eosinophilia, mast cell degranulation, chemokine (C-C motif) receptor 3 (CC receptor 3) and chemokine (C-X-C motif) receptor 1 (CXC receptor 1) gene expression amounts in the lungs, but restored the percentage of monocytes/macrophages. Our results suggest that menthone supplementation may alleviate allergic asthma through regulating airway allergic inflammation, protein overproduction, eosinophils infiltration, Th1/Th2 immune balance, CC receptor 3 and CXC receptor 1 gene expression amounts in the lungs but restoring the percentage of monocytes/macrophages in allergic asthmatic mice.

Menthone Inhalation Alleviates Local and Systemic Allergic Inflammation in Ovalbumin-Sensitized and Challenged Asthmatic Mice.

Menthone is rich in Mentha × Piperita L. essential oil and it has anti-inflammatory properties; research shows that it is useful, via percutaneous absorption, in treating inflammation-related diseases. However, anti-allergic inflammatory effects of volatile menthone have not yet been used to treat allergic asthma, in vivo. We hypothesized that menthone inhalation may have anti-inflammatory and anti-allergic effects in patients with allergic asthma. Therefore, in our study, menthone inhalation was used to treat ovalbumin (OVA)-sensitized and challenged asthmatic mice. Allergic inflammation mediator changes in the lungs and airways, sera, splenocytes, and peritoneal macrophages of the mice were measured. Relative expression amounts of six receptor genes related to allergic inflammation of the lungs and airways were quantitated using a two-step real time quantitative polymerase chain reaction (qPCR). Results showed that menthone inhalation increased serum OVA-specific IgG2a/IgG1 and IgG2a/IgE ratios, increased Th1-type cytokine production in the bronchoalveolar lavage fluid, and decreased nitric oxide, protein, and eotaxin levels. Menthone inhalation inhibited mast cell and eosinophil degranulation, and chemokine (C-C motif) receptor 3 (Ccr3) gene expression amounts, but (relatively) increased Th1 cytokine secretion by splenocytes. Our results evidence that menthone inhalation alleviates local and systemic allergic inflammation in asthmatic mice.

Active Ingredients from Euodia ruticarpa Steam Distilled Essential Oil Inhibit PC-3 Prostate Cancer Cell Growth via Direct Action and Indirect Immune Cells Conditioned Media In Vitro.

Active constituents isolated from Euodia ruticarpa (ER) steam distilled essential oil (SDEO) against PC-3 prostate cancer cell growth remain unclear. To clarify the puzzle, ER SDEO was extracted and further resolved into six isolated fractions ERF1-F6 with Sephadex LH-20 gel filtration chromatography to analyze their biological activities. Active ingredients in the isolated fractions were analyzed with GC-MS. Potential isolated fractions were selected to treat PC-3 cells with direct action and indirect treatment by mouse splenocyte- (SCM) and macrophage-conditioned media (MCM). The relationship between PC-3 cell viabilities and corresponding total polyphenols, flavonoid contents as well as Th1/Th2 cytokine profiles in SCM was analyzed using the Pearson product-moment correlation coefficient (r). As a result, ERF1-F3 was abundant in total polyphenols and flavonoids contents with diverse active ingredients. Treatments with ERF1-F3 at appropriate concentrations more or less inhibit PC-3 cell growth in a direct action manner. Only SCM, respectively, cultured with ER SDEO and ERF1-F3 markedly enhanced the effects to inhibit PC-3 cell growth, suggesting that secretions by splenocytes might involve anti-PC-3 effects. There are significantly negative correlations between PC-3 cell viabilities and IL-2, IL-10 as well as IL-10/IL-2 ratios in the corresponding SCM. Total polyphenol and flavonoid contents in the media cultured with ER SDEO isolated fractions positively correlated with IL-10 (Th2) and IL-10/IL-2 (Th2/Th1) cytokine secretion ratios by splenocytes, indicating that polyphenol and flavonoid components in ER SDEO isolated fractions promote Th2-polarized and anti-inflammatory characteristics. These new findings concluded that the inhibitory effects against PC-3 prostate cancer cell growth are attributed to active anti-inflammatory ingredients in ER SDEO and its active ERF1-F3 fractions through direct action and indirect treatment by modulating splenocytes' cytokine secretion profiles.

Rutaecarpine administration inhibits cancer cell growth in allogenic TRAMP-C1 prostate cancer mice correlating with immune balance in vivo.

BACKGROUND: Rutaecarpine (Rut) is a plant alkaloid abundant in Euodia ruticarpa which is a Chinese herbal medicine used for treating various cancers. However, the Rut administration effect on prostate cancer in vivo remains unclear. AIM: In the present study we established an allogenic TRAMP-C1 prostate cancer mouse model to evaluate the Rut administration effect and mechanism in vivo. METHODS: To unravel the Rut administration effect on prostate cancer in vivo, C57BL/6J male mice (8 weeks old) were randomly grouped (n = 9), subcutaneously loaded with TRAMP-C1 prostate cancer cells and immediately given daily by gavage with Rut dissolved in soybean oil at 7 mg (low dose), 35 mg (medium dose), and 70 mg/kg b.w./day (high dose) for successive 39 days. RESULTS: Rut administration significantly and dose-dependently reduced both tumor volume and solid prostate cancer weight in allogenic TRAMP-C1 male mice. Rut administration markedly increased (TNF-α+IFN-γ) (Th1-)/IL-10 (Th2-) cytokine secretion ratios by splenocytes and TNF-α (M1-)/IL-10 (M2-) cytokine secretion ratios by macrophages as compared to those of dietary control group, suggesting that Rut administration in vivo regulates the immune balance toward Th1- and M1-polarized characteristics. Decreased CD19+, CD4+ and CD8+ lymphocytes in the peripheral blood of allogenic TRAMP-C1 mice were significantly elevated by Rut administration. Tumor weights positively correlated with TNF-α secretions by splenocytes, suggesting that there is a tumor cachexia in the tumor-bearing mice. Tumor weights negatively correlated with IgG (Th1-antibody) levels in the sera, suggesting that Th1-polarized immune balance may inhibit prostate cancer cell growth. CONCLUSIONS: Our results evidenced that Rut administration suppresses prostate cancer cell growth in mice subcutaneously loaded with TRAMP-C1 cells and correlated the anti-cancer effects with Th1-polarized immune balance in vivo.

Pharmacological Effects of Guava (Psidium guajava L.) Seed Polysaccharides: GSF3 Inhibits PC-3 Prostate Cancer Cell Growth through Immunotherapy In Vitro.

The inhibitory effects of purified fractions isolated from guava seed polysaccharides (GSPS) including guava seed polysaccharide fraction 1 (GSF1), GSF2, and GSF3 on prostate cancer cells remain unclear. To clarify the anti-prostate cancer potential, GSPS, GSF1, GSF2, and GSF3 were isolated using Sepharose 6B gel filtration chromatography to assay their inhibitory effects on prostate PC-3 cell growth with direct action or indirect immunotherapy using either splenocyte conditioned media (SCM) or macrophage conditioned media (MCM). Correlations between cytokine profiles in the conditioned media and pro-apoptotic gene expression levels in the corresponding treated PC-3 cells were analyzed. Results showed that GSPS, GSF1, GSF2, and GSF3, particularly GSF3, through either direct action or indirect treatments using SCM or MCM, significantly (p < 0.05) inhibited PC-3 cell growth. GSF3 direct treatments increased pro-apoptotic Bax/anti-apoptotic Bcl-2 mRNA expression ratios in corresponding treated PC-3 cells. Either SCM or MCM cultured with GSF3 increased Fas mRNA expression levels in corresponding treated PC-3 cells. Both Th2-polarized and anti-inflammatory cytokine IL-10 either secreted in SCM or MCM were positively correlated with Fas mRNA expression levels in corresponding treated PC-3 cells. Our results suggest that GSF3 is a potent biological response modifier to decrease PC-3 cell growth through inducing apoptosis.

GSF3, a polysaccharide from guava (Psidium guajava L.) seeds, inhibits MCF-7 breast cancer cell growth via increasing Bax/Bcl-2 ratio or Fas mRNA expression levels.

Guava seed polysaccharide fraction 3 (GSF3) is an immunomodulatory polysaccharide from guava (Psidium guajava L.) seed polysaccharides. However, effects of GSF3 on the growth of breast cancer cells were not understood, yet. To clarify the GSF3 effects on breast cancer cell growth, GSF3 was subjected to treat MCF-7 cells using direct action or indirect immunotherapy using immune cells conditioned media, respectively. The viabilities of MCF-7 cells were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Changes in pro-(Bax)/anti-apoptotic (Bcl-2) and Fas mRNA expression levels in the treated MCF-7 cells were measured using two-step reverse transcription quantitative polymerase chain reaction. Our results showed that GSF3 inhibited MCF-7 cell growth through either direct action or indirect immunotherapy. GSF3 direct action significantly (P < 0.05) decreased Bcl-2 mRNA expression amount but increased pro-(Bax)/anti-apoptotic (Bcl-2) mRNA expression ratios in the treated cells. The splenocytes conditioned media cultured with GSF3 increased Fas mRNA expression amounts in the treated MCF-7 cells. There was a significant negative correlation between Th2-polarized cytokines secreted by immune cells and Fas mRNA expression levels in the corresponding treated MCF-7 cells. Our findings suggested that GSF3 is a potent anti-cancerous polysaccharide by direct action or indirectly modulating immune cell cytokine secretion profiles.

M1 Polarization but Anti-LPS-Induced Inflammation and Anti-MCF-7 Breast Cancer Cell Growth Effects of Five Selected Polysaccharides.

Five potential polysaccharides from guava seed (GSPS), common buckwheat (CBPS), bitter buckwheat (BBPS), red Formosa lambsquarters (RFLPS), and yellow Formosa lambsquarters (YFLPS) were selected to measure their effects on mouse peritoneal macrophages in the absence or presence of lipopolysaccharide (LPS). Macrophage-conditioned media (MCM) in the absence or presence of 5 selected polysaccharides were prepared to treat MCF-7 cells. The cell viability was determined using 3-(4,5-dimethylthiazol-2-diphenyl)-2,5-tetrazolium bromide (MTT) assay. Proinflammatory (also known as M1 type) (interleukin- (IL-) 1ß, IL-6 and tumor necrosis factor- (TNF-) α) and anti-inflammatory (also known as M2 type) (IL-10) cytokines secreted by macrophages were determined using ELISA. The relationship between MCF-7 cell growth and M1/M2 cytokine secretion profiles in the corresponding MCM were delineated. The results showed that 5 selected polysaccharides, except BBPS, significantly (P < 0.05) and dose-dependently increased M1 (IL-1ß + IL-6 + TNF-α)/M2 (IL-10) cytokine secretion ratios by macrophages in the absence of LPS, suggesting that four selected polysaccharides have M1 polarization property. However, all of 5 selected polysaccharides significantly (P < 0.05) decreased proinflammatory (IL-1ß + IL-6 + TNF-α)/anti-inflammatory (IL-10) cytokine secretion ratios by LPS-stimulated macrophages, exhibiting that all of the 5 selected polysaccharides, particularly GSPS, have anti-inflammatory potential. All MCM prepared with these selected polysaccharides (except YFLPS) significantly enhanced their inhibitory effects on MCF-7 cell growth. A negative correlation was noted between MCF-7 cell viabilities and M1/M2 cytokine secretion ratios ((IL-6 + TNF-α)/IL-10) in the corresponding MCM, suggesting that increases in M1 macrophages in the tumor microenvironment might inhibit MCF-7 cell growth. Particular polysaccharides including RFLPS, GSPS, YFLPS, and CBPS may increase the percentage of M1 macrophages in the tumor environment and further inhibit MCF-7 cell growth via immunotherapy.

Characterization of guava (Psidium guajava Linn) seed polysaccharides with an immunomodulatory activity.

To clarify the property of a novel guava seed polysaccharide (GSPS), GSPS was subjected to purify using Sepharose 6B gel filtration chromatography and further characterize the property of each individual isolated fraction. GSPS further resolved into three purified fractions, guava seed polysaccharide fraction 1 (GSF1), GSF2 and GSF3. Isolated GSF1, GSF2 and GSF3 were respectively subjected to high performance size exclusion chromatography; molecular weights of three polysaccharide fractions were determined. GSPS, GSF1, GSF2 and GSF3 were suggested to be proteopolysaccharides or glycoproteins. GSPS, GSF1, GSF2 and GSF3, particularly GSF3, were found to have a Th2-inclination property and anti-inflammatory potential. Heated GSF3 did not significantly (P > .05) decreased its immunomodulatory activity, suggesting that GSF3 is a proteopolysaccharide. The deproteinated GSF3 markedly lost its immunomodulatory activity, suggesting that both protein and carbohydrate moiety in GSF3 are essential to its immunomodulatory function. Analyses of monosaccharides composition in GSF3 using a pre-column derivatization high performance liquid chromatography exhibited that GSF3 was composed of glucuronic acid (3.28%), galacturonic acid (28.13%), galactose (14.88%), mannose (3.96%), glucose (22.99%), arabinose (7.31%), ribose (1.55%), xylose (14.81%), fucose (1.68%) and rhamnose (1.43%). Overall, we evidence that GSF3 is a low molecular weight proteopolysaccharide with potent anti-inflammatory and immunomodulatory effects.

The anti-TH17 polarization effect of Indigo naturalis and tryptanthrin by differentially inhibiting cytokine expression.

ETHNOPHARMACOLOGICAL RELEVANCE: The Chinese herbal medicine Qing-Dai (also known as Indigo naturalis) extracted from indigo-bearing plants including Baphicacanthus cusia (Ness) Bremek was previously reported to exhibit anti-psoriatic effects in topical treatment. TH17 was later established as a key player in the pathogenesis of psoriasis. We investigated the anti-TH17 effect of Indigo naturalis and its active compounds. The aim of this study is to evaluate the toxicity of Indigo naturalis (IN) and its derivatives on five cell types involved in psoriasis, and to study the anti-inflammatory mechanism for the toxicity. MATERIALS AND METHODS: Following the fingerprint and quantity analysis of indirubin, indigo, and tryptanthrin in IN extract, we used MTS kits to measure the anti-proliferative effect of IN and three active compounds on five different cell types identified in psoriatic lesions. Quantitative RT-PCR analysis was used to measure the expression of various genes identified in the activated keratinocytes and TH17 polarized gene expression in RORγt-expressing T cells. RESULTS: We showed that IN differentially inhibited the proliferation of keratinocytes and endothelial cells but not monocytes, fibroblasts nor Jurkat T cells. Among three active compounds identified in IN, tryptanthrin was the most potent compound to reduce their proliferation. In addition to differentially reducing IL6 and IL8 expression, both IN and tryptanthrin also potently decreased the expression of anti-microbial S100A9 peptide, CCL20 chemokine, IL1B and TNFA cytokines, independent of NF-κB-p65-activation. Their attenuating effect was also detected on the expression of signature cytokines or chemokines induced during RORγT-induced TH17 polarization. CONCLUSIONS: We were the first to confirm a direct anti-TH17 effect of both IN herbal extract and tryptanthrin.

Acorus gramineusand and Euodia ruticarpa Steam Distilled Essential Oils Exert Anti-Inflammatory Effects Through Decreasing Th1/Th2 and Pro-/Anti-Inflammatory Cytokine Secretion Ratios In Vitro.

To clarify the effects of steam distilled essential oils (SDEO) from herbs used in traditional Chinese medicine on immune functions, two potential herbs, Acorus gramineusand (AG) and Euodia ruticarpa (ER) cultivated in Taiwan, were selected to assess their immunomodulatory effects using mouse primary splenocytes and peritoneal macrophages. T helper type 1 lymphocytes (Th1) (IL-2), Th2 (IL-5), pro-inflammatory (TNF-α) and anti-inflammatory (IL-10) cytokines secreted by correspondent immune cells treated with SDEO samples were determined using enzyme-linked immunosorbent assay. The total amounts of potential phytochemicals, including total flavonoids, polyphenols and saponins, in these two selected SDEOs were measured and correlated with cytokine levels secreted by immune cells. Our results evidenced that ER SDEO is rich in total flavonoids, polyphenols and saponins. Treatments with AG and ER SDEO significantly (p < 0.05) increased IL-5/IL-2 (Th2/Th1) cytokine secretion ratios by splenocytes, suggesting that both AG and ER SDEO have the Th2-polarization property and anti-inflammatory potential. In addition, AG and ER SDEO, particularly ER SDEO, markedly decreased TNF-α/IL-10 secretion ratios by macrophages in the absence or presence of lipopolysaccharide (LPS), exhibiting substantial effects on spontaneous and LPS-induced inflammation. Significant correlations were found between the total polyphenols, flavonoids or saponins content in the two selected SDEOs and Th1/Th2 immune balance or anti-inflammatory ability in linear, non-linear or biphasic manners, respectively. In conclusion, our results suggest that AG and ER, particularly ER, SDEO have immunomodulatory potential in shifting the Th1/Th2 balance toward Th2 polarization in splenocytes and inhibiting inflammation in macrophages in the absence or presence of LPS.

Immune Cell-Conditioned Media Suppress Prostate Cancer PC-3 Cell Growth Correlating With Decreased Proinflammatory/Anti-inflammatory Cytokine Ratios in the Media Using 5 Selected Crude Polysaccharides.

Five different crude polysaccharides from guava seed (GSPS), bitter buckwheat (BBPS), common buckwheat (CBPS), red Formosa lambsquarters (RFLPS), and yellow Formosa lambsquarters (YFLPS) were isolated to treat human prostate cancer PC-3 cells via direct action or tumor immunotherapy. The splenocyte- and macrophage-conditioned media (SCM and MCM) were prepared using individual selected polysaccharides, and then SCM or MCM was further collected to treat PC-3 cells. The relationship between PC-3 cell growth and Th1/Th2 cytokines in SCM as well as proinflammatory/anti-inflammatory cytokine secretion profiles in MCM were delineated. The results showed that all 5 selected polysaccharides did not significantly inhibit PC-3 cell growth via direct action. However, SCM or MCM cultured in the absence or presence of 5 selected polysaccharides significantly (P < .05) inhibited PC-3 cell growth. MCM cultured with 5 polysaccharides dose dependently enhanced their inhibitory effects on the viabilities of PC-3 cells than those cultured without polysaccharides. There was a significant (P < .05) negative correlation between PC-3 cell viabilities and (interleukin [IL]-6 + tumor necrosis factor [TNF]-α)/IL-10 level ratios in the corresponding MCM, implying that macrophages suppress PC-3 cell growth through decreasing secretion ratios of proinflammatory/anti-inflammatory cytokines in a tumor microenvironment.

Quercetin intraperitoneal administration ameliorates lipopolysaccharide-induced systemic inflammation in mice.

AIMS: The purpose of this study was to unravel pharmacological effects of quercetin (Q) on systemic inflammation in septic mice, and compare it to quercetin-3-glucuronide (Q3G), a major metabolite of Q. MAIN METHODS: A suitable sepsis mouse model was first established using lipopolysaccharide (LPS) injected intraperitoneally (i.p.). Q or Q3G was administered i.p. to septic mice in a prophylactic or therapeutic manner. Pro-inflammatory (TNF-α, IL-1ß and IL-6) and anti-inflammatory (IL-10) cytokine secretion profiles by peritoneal macrophages of the mice were measured using ELISA. KEY FINDINGS: Mice which received 8mg/kg BW LPS i.p. for 12h resulted in intermediate systemic inflammation, suggesting a useful mild septic mouse model. At non-toxic doses, Q or Q3G (0.06 or 0.15µmol/mouse) i.p. injected in a prophylactic manner significantly (P<0.05) increased anti-inflammatory IL-10 secretions by peritoneal macrophages of the LPS-induced septic mice. Q, but not Q3G, i.p. injected in a therapeutic manner significantly (P<0.05) increased IL-10 secretions by peritoneal macrophages of the septic mice. SIGNIFICANCE: Our data suggest that Q, but not Q3G, has pharmacological effects to ameliorate systemic inflammation. These results are the first to show that Q has potent potential against sepsis in both prophylactic and therapeutic manners in vivo.

Farnesol, a sesquiterpene alcohol in herbal plants, exerts anti-inflammatory and antiallergic effects on ovalbumin-sensitized and -challenged asthmatic mice.

To investigate the effect of farnesol on allergic asthma, three farnesol doses were extra-added into AIN-76 feed consumed by ovalbumin- (OVA-) sensitized and -challenged mice continuously for 5 weeks, at approximately 5, 25, and 100 mg farnesol/kg, BW/day. The results showed that there were no significant differences in body weight, feed intake, and visceral organ weights between the farnesol supplementation and dietary control groups. Farnesol supplementation decreased interleukin (IL)-6/IL-10 level ratios in bronchoalveolar lavage fluid (BALF). Farnesol supplementation significantly (P < 0.05) restored the cytokine secretion ability of peritoneal macrophages that was suppressed as a result of OVA sensitization and challenge and slightly decreased tumor necrosis factor (TNF-α)/IL-10 cytokine secretion ratios. Farnesol supplementation slightly (P > 0.05) decreased IL-4 but significantly (P < 0.05) increased IL-2 levels secreted by the splenocytes in the presence of OVA, implying that farnesol might have a systemic antiallergic effect on allergic asthmatic mice. Farnesol supplementation significantly (P < 0.05) increased IL-10 levels secreted by the splenocytes in the presence of OVA, suggesting that farnesol might have an anti-inflammatory potential to allergic asthmatic mice. Overall, our results suggest that farnesol supplementation may be beneficial to improve the Th2-skewed allergic asthmatic inflammation.

Quercetin uptake and metabolism by murine peritoneal macrophages in vitro.

Quercetin (Q), a bioflavonoid ubiquitously distributed in vegetables, fruits, leaves, and grains, can be absorbed, transported, and excreted after oral intake. However, little is known about Q uptake and metabolism by macrophages. To clarify the puzzle, Q at its noncytotoxic concentration (44µM) was incubated without or with mouse peritoneal macrophages for different time periods. Medium alone, extracellular, and intracellular fluids of macrophages were collected to detect changes in Q and its possible metabolites using high-performance liquid chromatography. The results showed that Q was unstable and easily oxidized in either the absence or the presence of macrophages. The remaining Q and its metabolites, including isorhamnetin and an unknown Q metabolite [possibly Q- (O-semiquinone)], might be absorbed by macrophages. The percentage of maximal Q uptake by macrophages was found to be 2.28% immediately after incubation; however, Q uptake might persist for about 24 hours. Q uptake by macrophages was greater than the uptake of its methylated derivative isorhamnetin. As Q or its metabolites entered macrophages, those compounds were metabolized primarily into isorhamnetin, kaempferol, or unknown endogenous Q metabolites. The present study, which aimed to clarify cellular uptake and metabolism of Q by macrophages, may have great potential for future practical applications for human health and immunopharmacology.

Quercetin, but not its metabolite quercetin-3-glucuronide, exerts prophylactic immunostimulatory activity and therapeutic antiinflammatory effects on lipopolysaccharide-treated mouse peritoneal macrophages ex vivo.

This study investigated the prophylactic or therapeutic effects of quercetin (Q) and its metabolite quercetin-3-glucuronide (Q3G) on lipopolysaccharide (LPS)-induced inflammation in mouse peritoneal macrophages ex vivo. Changes in pro- and antiinflammatory cytokine secretion profiles were determined. The results showed that Q or Q3G in vitro treatments lower than 50 µM did not exhibit cytotoxicity on macrophages. At noncytotoxic doses, Q and Q3G, particularly Q, administration in a prophylactic ex vivo model increased pro-/antiinflammatory cytokine secretion ratios by macrophages in the absence or presence of LPS. Quercetin, but not Q3G, administration in a therapeutic ex vivo model decreased pro-/antiinflammatory cytokine secretion ratios in the absence or presence of LPS. Our results indicated that Q and Q3G administrations in a prophylactic manner might act as an immunostimulatory agent, but Q presented better ability than Q3G. Quercetin might have a therapeutic, but not prophylactic, effect on spontaneous or LPS-induced inflammation in vivo.