Houttuynia cordata Thunb. Volatile Oil Exhibited Anti‐inflammatory Effects In Vivo and Inhibited Nitric Oxide and Tumor Necrosis Factor‐α Production in LPS‐stimulated Mouse Peritoneal Macrophages In Vitro

Houttuynia cordata Thunb. (HC) is a medicinal herb that generally used in traditional Chinese medicine for treating allergic inflammation. The present study investigated the inhibitory effect of the volatile oil from HC Thunb. on animal models of inflammation and the production of inflammatory mediators in vivo and in vitro. In vivo, xylene‐induced mouse ear edema, formaldehyde‐induced paw edema and carrageenan‐induced mice paw edema were significantly decreased by HC volatile oil. HC volatile oil showed pronounced inhibition of prostaglandin (PG) E₂ and malondialdehyde production in the edematous exudates. In vitro exposure of mouse resident peritoneal macrophages to 1, 10, 100 and 1000 µg/mL of HC volatile oil significantly suppressed lipopolysaccharide (LPS)‐stimulated production of NO and tumor necrosis factor‐α (TNF‐α) in a dose‐dependent manner. Exposure to HC volatile oil had no effect on cell viability and systemic toxicity. Furthermore, HC volatile oil inhibited the production of NO and TNF‐α by down‐regulating LPS‐stimulated iNOS and TNF‐α mRNA expression. Western blot analysis showed that HC volatile oil attenuated LPS‐stimulated synthesis of iNOS and TNF‐α protein in the macrophages, in parallel. These findings add a novel aspect to the biological profile of HC and clarify its anti‐inflammatory mechanism. Copyright © 2012 John Wiley & Sons, Ltd.     

Inhibition of LPS‐Induced TNF‐α and NO Production in Mouse Macrophage and Inflammatory Response in Rat Animal Models by a Novel Ayurvedic Formulation,BV‐9238

Rheumatoid arthritis is a chronic crippling disease, where protein‐based tumor necrosis factor‐alpha (TNF‐α) inhibitors show significant relief, but with potentially fatal side effects. A need for a safe, oral, cost‐effective small molecule or phyto‐pharmaceutical is warranted. BV‐9238 is an Ayurvedic poly‐herbal formulation containing specialized standardized extracts of Withania somnifera, Boswellia serrata, Zingiber officinale and Curcuma longa. The anti‐inflammatory and anti‐arthritic effects of BV‐9238 were evaluated for inhibition of TNF‐α and nitric oxide (NO) production, in lipopolysaccharide‐stimulated, RAW 264.7, mouse macrophage cell line. BV‐9238 reduced TNF‐α and NO production, without any cytotoxic effects. Subsequently, the formulation was tested in adjuvant‐induced arthritis (AIA) and carrageenan‐induced paw edema (CPE) rat animal models. AIA was induced in rats by injecting Freund's complete adjuvant intra‐dermally in the paw, and BV‐9238 and controls were administered orally for 21 days. Arthritic scores in AIA study and inflamed paw volume in CPE study were significantly reduced upon treatment with BV‐9238. These results suggest that the anti‐inflammatory and anti‐arthritic effects of BV‐9238 are due to its inhibition of TNF‐α, and NO, and this formulation shows promise as an alternate therapy for inflammatory disorders where TNF‐α and NO play important roles. Copyright © 2014 John Wiley & Sons, Ltd.     

Praeruptorin a inhibits lipopolysaccharide‐induced inflammatory response in murine macrophages through inhibition of NF‐κB pathway activation

Praeruptorin A (PA) is a pyranocoumarin compound isolated from the dried root of Peucedanum praeruptorum Dunn (Umbelliferae). However, the antiinflammatory effect of PA has not been reported. The present study investigated the antiinflammatory effect of PA in lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophage cells. PA significantly inhibited the LPS‐induced production of nitric oxide (NO), interleukin‐1β (IL‐1β) and tumor necrosis factor‐α (TNF‐α). The mRNA and protein expressions of inducible nitric oxide synthase (iNOS), IL‐1β and TNF‐α were also suppressed by this compound. Further study showed that PA decreased the cytoplasmic loss of inhibitor κB‐α (IκB‐α) protein and inhibited the translocation of NF‐κB from cytoplasm to nucleus. Taken together, the results suggest that PA may exert antiinflammatory effects in vitro in LPS‐stimulated RAW 264.7 macrophages through inhibition of NF‐κB signal pathway activation. Copyright © 2010 John Wiley & Sons, Ltd.     

Inhibition of Proinflammatory Biomarkers in THP1 Macrophages by Polyphenols Derived From Chamomile,Meadowsweet and Willow bark

Antiinflammatory compounds in the diet can alleviate excessive inflammation, a factor in the pathogenesis of common diseases such as rheumatoid arthritis, atherosclerosis and diabetes. This study examined three European herbs, chamomile (Matricaria chamomilla), meadowsweet (Filipendula ulmaria L.) and willow bark (Salix alba L.), which have been traditionally used to treat inflammation and their potential for use as antiinflammatory agents. Aqueous herbal extracts and isolated polyphenolic compounds (apigenin, quercetin and salicylic acid, 0–100 μM) were incubated with THP1 macrophages, and interleukin (IL)‐1β, IL‐6 and tumour necrosis factor‐alpha (TNF‐α) were measured. At concentrations of 10 μM, both apigenin and quercetin reduced IL‐6 significantly ( p < 0.05). Apigenin at 10 μM and quercetin at 25 μM reduced TNF‐α significantly ( p < 0.05). Amongst the herbal extracts, willow bark had the greatest antiinflammatory activity at reducing IL‐6 and TNF‐α production. This was followed by meadowsweet and then chamomile. The lowest effective antiinflammatory concentrations were noncytotoxic (MTT mitochondrial activity assay). The Comet assay, which was used to study the protective effect of the isolated phenols against oxidative damage, showed positive results for all three polyphenols. These are the first findings that demonstrate the antiinflammatory capacity of these herbal extracts. Copyright © 2012 John Wiley & Sons, Ltd.     

Anti‐inflammatory effect of isopimarane diterpenoids from Kaempferia galanga

Two new isopimarane diterpenes, 1α‐hydroxy‐14α‐methoxyisopimara‐8(9),15‐diene ( 7 ) and 1α,14α‐dihydroxyisopimara‐8(9),15‐diene ( 9 ) and eight known isopimarane diterpenes including (‐)‐sandaracopimaradiene ( 1 ), 6β‐acetoxysandaracopimaradiene‐9α‐ol ( 2 ), sandaracopimaradiene‐7β,9α‐diol ( 3 ), sandaracopimaradiene‐1α,9α‐diol ( 4 ), 6β‐acetoxysandaracopimaradiene‐9α‐ol‐1‐one ( 5 ), 6β‐acetoxysandaracopimaradiene‐1α,9α‐diol ( 6 ), 6β,14α‐dihydroxyisopimara‐8(9),15‐diene ( 8 ), and 6β,14β‐dihydroxyisopimara‐8(9),15‐diene ( 10 ) were isolated from hexane fraction of Kaempferia galanga ethanol extract. Compounds 5 , 6 , 8 , and 9 exerted the good anti‐inflammatory effect on lipopolysaccharide‐stimulated nitric oxide production from RAW264.7 cells with IC₅₀ of 11.2, 7.7, 14.3, and 12.1 μM, respectively. These four compounds inhibited nitric oxide synthase (iNOS) mRNA expression. Compounds 5 and 6 also suppressed cyclooxygenase 2 (COX‐2) mRNA expression; in addition, compound 6 had mild inhibitory effect on TNF‐α mRNA. Among these compounds, 5 dramatically inhibited iNOS and COX‐2 mRNA expression. The influential structures were proposed to be oxygen substitute at C‐1, C‐6, and α‐OH at C‐14.     

Anti‐TNF‐α Activity of Brazilian Medicinal Plants and Compounds from Ouratea semiserrata

Several plant species are used in Brazil to treat inflammatory diseases and associated conditions. TNF‐α plays a pivotal role on inflammation, and several plant extracts have been assayed against this target, both in vitro and in vivo. The effect of 11 Brazilian medicinal plants on TNF‐α release by LPS‐activated THP‐1 cells was evaluated. The plant materials were percolated with different solvents to afford 15 crude extracts, whose effect on TNF‐α release was determined by ELISA. Among the evaluated extracts, only Jacaranda caroba (Bignoniaceae) presented strong toxicity to THP‐1 cells. Considering the 14 non‐toxic extracts, TNF‐α release was significantly reduced by seven of them (inhibition > 80%), originating from six plants, namely Cuphea carthagenensis (Lythraceae), Echinodorus grandiflorus (Alismataceae), Mansoa hirsuta (Bignoniaceae), Ouratea semiserrata (Ochnaceae), Ouratea spectabilis and Remijia ferruginea (Rubiaceae). The ethanol extract from O. semiserrata leaves was fractionated over Sephadex LH‐20 and RP‐HPLC to give three compounds previously reported for the species, along with agathisflavone and epicatechin, here described for the first time in the plant. Epicatechin and lanceoloside A elicited significant inhibition of TNF‐α release, indicating that they may account for the effect produced by O. semiserrata crude extract. Copyright © 2015 John Wiley & Sons, Ltd.     

The anti‐inflammatory effect of baicalin on hypoxia/reoxygenation and TNF‐α induced injury in cultural rat cardiomyocytes

The aim of present study was to investigate the effect of baicalin on hypoxia/reoxygenation (H/R) injury in cardiomyocytes and the mechanisms involved, particularly in relation to cytokines. The cardiomyocytes for the H/R groups were placed into a hypoxic chamber for 12 h and then underwent reoxygenation for 1 h. The cells in the TNF‐α groups were administered 100 ng/mL rrTNF‐α and incubated for 13 h under normal conditions. The cells in the baicalin pretreatment groups were administered 10 μM baicalin 30 min prior to exposure to H/R or TNF‐α. It was observed that pretreatment with baicalin (10 μM) significantly reduced the cell damage and death induced by H/R or TNF‐α. In the culture medium of the H/R cells, the SOD activity increased, while TNF‐α was decreased by baicalin. The levels of IL‐6 in culture medium for H/R or TNF‐α treated cells were suppressed by baicalin pretreatment. In contrast, the levels of IL‐10 in culture medium for H/R or TNF‐α treated cells were significantly elevated by baicalin. Moreover, baicalin inhibited the nuclear translocation of NF‐κB induced by H/R or TNF‐α. In conclusion, baicalin may protect cardiomyocytes from H/R injury through an anti‐inflammatory mechanism. Copyright © 2009 John Wiley & Sons, Ltd.     

Rhynchophylline Attenuates LPS‐induced Pro‐inflammatory Responses through Down‐regulation of MAPK/NF‐κB Signaling Pathways in Primary Microglia

Excessive activation of microglial cells has been implicated in various types of neuroinflammation. Suppression of microglial activation would have therapeutic benefits, leading to the alleviation of the progression of neurodegeneration. In this study, the inhibitory effects of rhynchophylline (RIN), a tetracyclic oxindole alkaloid component isolated from Uncaria rhynchophylla (Miq.) Jacks., on the production of pro‐inflammatory mediators were investigated in lipopolysaccharide (LPS)‐stimulated microglia. The results showed that RIN markedly reduced the production of nitric oxide (NO), prostaglandins E₂ (PGE₂), monocyte chemoattractant protein (MCP‐1), tumor necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) in LPS‐activated microglia. The mRNA expression levels of iNOS and COX‐2 were also depressed by RIN in a concentration‐dependent manner. Further studies revealed that RIN blocked IκBα phosphorylation and degradation, inhibited the phosphorylation of mitogen‐activated protein kinases (MAPKs). In summary, these data suggest that RIN suppresses inflammatory responses of microglia and may act as a potential therapeutic agent for various neurodegenerative diseases involving neuroinflammation. Copyright © 2012 John Wiley & Sons, Ltd.     

Anti‐Inflammatory Sesquiterpene Lactones from Lychnophora trichocarpha Spreng. (Brazilian Arnica)

The aerial parts of Lychnophora trichocarpha Spreng. (Asteraceae) are used macerated in water or ethanol to treat inflammation, pain, rheumatism, contusions, bruises and insect bites in Brazilian traditional medicine. In this study, anti‐inflammatory activity of ethanol extract from aerial parts of L. trichocarpha and its ethyl acetate fraction was investigated. Sesquiterpene lactones, lychnopholide (Lyc) and eremantholide C (EreC), isolated of ethyl acetate fraction, were also assayed for in vitro and in vivo anti‐inflammatory activity. Topical treatment with ointments containing ethanol extract, its ethyl acetate fraction and sesquiterpene lactones significantly reduced carrageenan‐induced mice paw oedema. In vitro assays demonstrated that Lyc inhibited interferon ‐γ/lipopolysaccharide ‐stimulated nitric oxide (NO) production in J774A.1 macrophages and increased production of IL‐10 anti‐inflammatory cytokine. The reduction of tumor necrosis factor‐α (TNF‐α) production by EreC was accompanied by an increased production of IL‐10 in a concentration‐dependent manner in J774A.1 macrophages. The anti‐inflammatory effect of Lyc seems to involve the inhibition of production of NO and increased production of IL‐10. The mechanism of the effect of EreC on the reduction of carrageenan‐induced paw oedema may be attributed to inhibition of production of TNF‐α and stimulation of IL‐10 production. The results corroborate the use of ethanol extract from Lychnophora trichocarpha in folk medicine for anti‐inflammatory action and indicate that the topical route is suitable for use. Copyright © 2012 John Wiley & Sons, Ltd.     

Prophylactic and Therapeutic Effects of Acanthopanax senticosus Harms Extract on Murine Collagen‐induced Arthritis

Evidences are accumulating that extract of Acanthopanax senticosus Harms (ASH; syn Eleutherococcus senticosus [Rupr. & Maxim.] Maxim), a shrub native to Northeastern Asia, has antiinflammatory effects. In this study, we examined prophylactic and therapeutic effects of ASH extract (ASHE) on rheumatoid arthritis using collagen‐induced arthritis (CIA) mouse model. Acanthopanax senticosus Harms extract was administered before the onset of arthritis in the prophylaxis model. In the therapeutic model, ASHE was administered after the onset of arthritis with or without anti‐TNF‐α antibody. The ASHE treatment showed efficacy before onset of CIA but there was no effect after CIA was established. The ASHE treatment delayed the onset and decreased severity of CIA. In vitro examinations showed that ASHE is an antioxidant and that ASHE suppresses TNF‐α and interleukin‐6 production in human peripheral blood mononuclear cells. The combination therapy with ASHE and anti‐TNF‐α antibody reduced the severity of arthritis compared with anti‐TNF‐α antibody alone. The present study shows that ASHE has prophylactic effect against CIA and support therapeutic effect of anti‐TNF‐α antibody. © 2014 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.     

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 © 2011 John Wiley & Sons, Ltd.     

The Effects of Low and High Concentrations of Luteolin on Cultured Human Endothelial Cells Under Normal and Glucotoxic Conditions: Involvement of Integrin‐Linked Kinase and Cyclooxygenase‐2

Luteolin protects against high glucose (HG)‐induced endothelial dysfunction whereas its cytotoxicity has been reported against normal endothelial cells. This study was undertaken to determine luteolin cytoprotective and cytotoxic dose ranges and to elucidate their respective mechanisms. Luteolin prevented HG‐induced human umbilical vein endothelial cell (HUVEC) death with an EC50 value of 2.0 ± 0.07 μM. The protective effect of luteolin was associated with decreased intracellular reactive oxygen species (ROS) and Ca²⁺ (Cai²⁺) levels and enhanced nitric oxide (NO) production. At high concentrations, luteolin caused HUVEC death in normal glucose (NG) and HG states (LC₅₀ 40 ± 2.23 and 38 ± 1.12 μM, respectively), as represented by increased ROS and Cai²⁺ and decreased NO. Western blots illustrated that exposure to HG increased cyclooxygenase‐2 (COX‐2) and integrin‐linked kinase (ILK) expression. Luteolin at low concentrations suppressed HG‐mediated up‐regulation of COX‐2 but maintained HG‐induced over‐expression of ILK while at high concentrations significantly increased COX‐2 and decreased ILK expression in both HG and NG states. Our data indicated that cytoprotective action of luteolin was manifested with much lower concentrations, by a factor of approximately 20, compared with cytotoxic activity under both normal or glucotoxic conditions. It appears that luteolin exerts its action, in part, by modulating ILK expression which is associated with regulation of COX‐2 expression and NO production in endothelial cells. Copyright © 2014 John Wiley & Sons, Ltd.     

Anti‐Ulcerative Colitis Activity of Compounds from Euphorbia granuleta Forssk

The aim of the present study was to evaluate the anti‐ulcerative colitis (UC) activity of the total alcohol extracts of Euphorbia granuleta Forssk. (Euphorpiaceae), isolate and identify the active compounds that could be responsible for the activity, in addition to determination of the possible mechanism of action. Six compounds were isolated and identified from this plant: three phenolic compounds (kampferol, kampferol‐3‐glucoside and kampferol‐3‐galactoside) in addition to three steroidal compounds (1‐ethoxypentacosane, heptacosan‐1‐ol and β‐sitosterol). Three compounds (heptacosan‐1‐ol, β‐sitosterol and kampferol‐3‐galactoside) were found to be responsible for the anti‐UC activity of E. granuleta extract. The anti‐UC activity of these compounds may be explained by reducing the pro‐inflammatory cytokine tumor necrosis factor‐alpha (TNF‐α), in addition to reduction of colonic malondialdehyde (MDA) contents. No side effects were reported on liver and kidney functions. The active compounds reduced both serum TNF‐α and mucosal MDA levels. Copyright © 2013 John Wiley & Sons, Ltd.     

Sour Cherry Seed Kernel Extract Increases Heme Oxygenase‐1 Expression and Decreases Representation of CD3+ TNF‐α + and CD3 + IL‐8+ Subpopulations in Peripheral Blood Leukocyte Cultures from Type 2 Diabetes Patients

The present study evaluates a hypothesis that sour cherry (Prunus cerasus) seed extracts (SCE) modulate CD3+ T lymphocyte activity in ways predictive of potential for uses of SCE in management of inflammatory diseases. Peripheral blood mononuclear cells (PBMC) from 12 type 2 diabetes (T2DM) patients and eight healthy control subjects were cultured 24 h with 100 ng/ml lipopolysaccharide (LPS) to increase inflammatory signaling and co‐incubated with 0.5–100 µg/ml SCE. Cultures were evaluated by two‐color flow cytometry for percent representation of CD3+ IL8+ and CD3 + TNF‐α + cells which express interleukin‐8 (IL‐8), and tumor necrosis factor‐α, (TNF‐α+) respectively, and by enzyme‐linked immunoassay for lymphocyte‐associated heme oxygenase‐1 (HO‐1, known to be induced by SCE). SCE dosage ranges of 0.5–100 µg/ml in cell cultures significantly suppressed LPS‐increased CD3 + TNF‐α + and CD3 + IL8+ representation from all participants (p < 0.05), with greater pharmacological effect noted in suppression of CD3 + TNF‐α + noted in cells from T2DM patients versus healthy control subjects. These effects correlated with increased HO‐1 expression in SCE‐treated PBMC from all subjects (p < 0.05). Since TNF‐α and IL‐8 are diagnostic/prognostic biomarkers for many inflammatory syndromes, the capacity of SCE to down‐regulate representation of cells that express them suggests potential for therapeutic use of SCE in T2DM and other diseases. Copyright © 2012 John Wiley & Sons, Ltd.     

Anti‐inflammatory effects of chemical components from Ginkgo biloba L. male flowers on lipopolysaccharide‐stimulated RAW264.7 macrophages

Ginkgo biloba L., well known as living fossil, have various pharmacological activities. Eighteen compounds were isolated from Ginkgo male flowers including a novel matsutake alcohol glycoside, Ginkgoside A ( 1 ), and 17 known compounds—calaliukiuenoside ( 2 ), benzylalcohol O‐α‐l ‐arabinopyranosyl‐(1 → 6)‐β‐d ‐glucopyranoside ( 3 ), amentoflavone ( 4 ), sciadopitysin ( 5 ), bilobetin ( 6 ), isoginkgetin ( 7 ), olivil 4‐O‐β‐d ‐glucopyranoside ( 8 ), dihydrodehydrodiconiferyl alcohol‐4‐O‐β‐d ‐glucoside ( 9 ), (+)‐cyclo‐olivil‐6‐O‐β‐d ‐glucopyranoside ( 10 ), (?)‐isolariciresinol 4‐O‐β‐d ‐glucopyranoside ( 11 ), coniferin ( 12 ), trans‐cinnamic acid‐4‐O‐β‐d ‐glucopyranoside ( 13 ), p‐coumaryl alchol glucoside ( 14 ), stroside B ( 15 ), methylconiferin ( 16 ), cis‐p‐coumaric acid 4‐O‐β‐d ‐glucopyranoside ( 17 ), and cis‐coniferin ( 18 ). Thirteen of these compounds had not previously found in Ginkgo. All extractive fractions and isolated compounds were evaluated for their anti‐inflammatory ability in the lipopolysaccharide‐induced RAW264.7 macrophages. The ethanol extract of Ginkgo flowers and the chloroform and ethyl acetate fractions can significantly decrease nitric oxide (NO), interleukin‐6 (IL‐6), and prostaglandin E₂ (PGE₂) production at 100 μg/ml. The most effective compounds, bilobetin ( 6 ) and isoginkgetin ( 7 ), elevated the NO inhibition ratios to 80.19% and 82.37% at 50 μM, respectively. They also exhibited significant dose‐dependent inhibitory effects on tumor necrosis factor‐α, IL‐6, PGE₂, inducible NO synthase mRNA, and cyclooxygenase‐2 mRNA levels. So they can be promising candidates for the development of new anti‐inflammatory agents.     

Myrislignan Attenuates Lipopolysaccharide‐induced Inflammation Reaction in Murine Macrophage Cells Through Inhibition of NF‐κB Signalling Pathway Activation

Myrislignan is a new kind of lignan isolated from Myristica fragrans Houtt. Its antiinflammatory effects have not yet been reported. In the present study, the antiinflammatory effects and the underlying mechanisms of myrislignan in lipopolysaccharide (LPS)‐induced inflammation in murine RAW 264.7 macrophage cells were investigated. Myrislignan significantly inhibited LPS‐induced production of nitric oxide (NO) in a dose‐dependent manner. It inhibited mRNA expression and release of interleukin‐6 (IL‐6) and tumour necrosis factor‐α (TNF‐α). This compound significantly inhibited mRNA and protein expressions of inducible NO synthase (iNOS) and cyclooxygenase‐2 (COX‐2) dose‐dependently in LPS‐stimulated macrophage cells. Further study showed that myrislignan decreased the cytoplasmic loss of inhibitor κB‐α (IκB‐α) protein and the translocation of NF‐κB from cytoplasm to the nucleus. Our results suggest that myrislignan may exert its antiinflammatory effects in LPS‐stimulated macrophages cells by inhibiting the NF‐κB signalling pathway activation. Copyright © 2012 John Wiley & Sons, Ltd.