Anti-inflammatory activity of ethanolic extract of Sargassum sagamianum in RAW 264.7 cells

Aim of the study is to evaluate the antiinflammatory effects of ethanolic extract of the marine brown alga Sargassum sagamianum collected from Yeonhwari coast of Korea. Ethanolic extract of S. sagamianum (SA-E extract) inhibited expression of nitric oxide (NO) and cytokines (IL-6, IL-1β, and TNF-α) as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 in lipopolysaccharide (LPS)-induced RAW 264.7 cells without affecting cell viability. In addition, the expression of nuclear factor (NF)-κB p65 was suppressed by SA-E extract. Furthermore, the rate of formation of edema in the mouse ear was reduced by 46% at the highest dose tested (250 mg/kg) compared to that in the control. This study suggests that SA-E extract exerts potent inhibitory effects on LPS-induced expression of inflammatory mediators such as NO, iNOS, COX-2, and cytokines in macrophages through suppression of the NF-κB p65 pathway. SA-E extract might have potential clinical applications as an anti-inflammatory agent.     

Polygonum viviparum L. inhibits the lipopolysaccharide‐induced inflammatory response in RAW264.7 macrophages through haem oxygenase‐1 induction and activation of the Nrf2 pathway

BACKGROUND: Polygonum viviparum L. (PV) is a member of the family Polygonaceae and is widely distributed in high‐elevation areas. It is used as a folk remedy to treat inflammation‐related diseases. This study was focused on the anti‐inflammatory response of PV against lipopolysaccharide (LPS)‐induced inflammation in RAW264.7 macrophages. RESULTS: Treatment with PV did not cause cytotoxicity at 0–50 µg mL^?1 in RAW264.7 macrophages, and the IC₅₀ value was 270 µg mL^?1. PV inhibited LPS‐stimulated nitric oxide (NO), prostaglandin (PG)E₂, interleukin (IL)‐1β and tumour necrosis factor (TNF)‐α release and inducible NO synthase (iNOS) and cyclooxygenase (COX)‐2 protein expression. In addition, PV suppressed the LPS‐induced p65 expression of nuclear factor (NF)‐κB, which is associated with the inhibition of IκB‐α degradation. These results suggest that, among mechanisms of the anti‐inflammatory response, PV inhibits the production of NO and these cytokines by down‐regulating iNOS and COX‐2 gene expression. Furthermore, PV can induce haem oxygenase (HO)‐1 protein expression through nuclear factor E2‐related factor 2 (Nrf2) activation. A specific inhibitor of HO‐1, zinc(II) protoporphyrin IX, inhibited the suppression of iNOS and COX‐2 expression by PV. CONCLUSION: These results suggest that PV possesses anti‐inflammatory actions in macrophages and works through a novel mechanism involving Nrf2 actions and HO‐1. Thus PV could be considered for application as a potential therapeutic approach for inflammation‐associated disorders. © 2012 Society of Chemical Industry     

Anti-inflammatory effect of flavonoids isolated from Korea Citrus aurantium L. on lipopolysaccharide-induced mouse macrophage RAW 264.7 cells by blocking of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signalling pathways

Citrus fruits are an abundant source of various flavonoids, which have been used as a traditional herbal medicine in Korea and China. Most flavonoids are known to have anti-oxidant, anti-bacterial and analgesic properties. In this study, it was examined whether flavonoids (nobiletin, naringin and hesperidin) isolated from Korea Citrus aurantium L. inhibited the pro-inflammatory mediators including cytokines by blocking nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signalling in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The flavonoids suppressed mRNA and protein expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in LPS-induced macrophages. The molecular mechanism was associated with inhibition of the degradation/phosphorylation of I-κB-α and nuclear translocation of the NF-κB p-65 as well as phosphorylation of MAPK by flavonoids. These results suggest that flavonoids have anti-inflammatory effects by suppressing expression of COX-2, iNOS and cytokines by blocking the NF-κB and MAPK signalling in RAW 264.7 macrophages.     

甘草多糖对小鼠腹腔巨噬细胞NO、iNOS及iNOS mRNA表达的影响

通过研究甘草多糖(GP)对小鼠腹腔巨噬细胞NO、iNOS 的生成及iNOS mRNA 表达的影响,结果表明：GP 能剂量依赖性地增加活化的巨噬细胞中NO 的生成量及iNOS 的活性,当GP 的添加浓度为400μg/ml 时,NO、iNOS 的分泌量和iNOS mRNA 表达达到最大。固定GP 添加量为100μg/ml,培养时间在48h 时,NO、iNOS 的分泌量和iNOS mRNA 表达量最大。这表明GP 刺激巨噬细胞NO 合成的调节可发生在iNOS 的转录水平,从而刺激巨噬细胞NO 大量合成与释放,GP 的免疫调节作用和抗肿瘤作用机制可能与GP 刺激巨噬细胞iNOS 从头合成从而增加NO 生成有关。     

大蒜素对脂多糖诱导腹腔巨噬细胞炎症反应的抑制作用及机制

目的:探讨大蒜素对脂多糖(LPS)诱导小鼠腹腔巨噬细胞炎症反应的抑制作用及机制。方法:建立LPS诱导小鼠腹腔巨噬细胞炎症反应细胞模型,并用地塞米松和不同浓度大蒜素处理,MTT法检测细胞活力,中性红吞噬实验检测吞噬能力,Griess法检测一氧化氮(NO)及ELISA法检测COX-2酶活性和IL-6的分泌,qPCR检测环氧合酶2(COX-2)、一氧化氮合酶(iNOS)和IL-6的mRNA表达水平,Western Blot检测COX-2、iNOS和IL-6的蛋白表达以及核转录因子NF-κB p65及其磷酸化产物的相对表达。结果:大蒜素浓度在40~160 μg/mL范围内对腹腔巨噬细胞均无细胞毒性;与LPS组比较,大蒜素处理组能促进腹腔巨噬细胞的吞噬能力,能显著(P<0.05)抑制炎症因子COX-2酶活性、NO和IL-6的分泌,能显著(P<0.05)抑制基因COX-2、iNOS和IL-6 mRNA和蛋白的相对表达,并极显著(P<0.01)抑制NF-κB p65信号通路的磷酸化。结论:大蒜素能显著抑制LPS诱导的小鼠腹腔巨噬细胞的炎症反应,其机制可能与抑制NF-κB信号通路激活有关。     

Comparative effects of tocotrienol-rich fraction, α-tocopherol and α-tocopheryl acetate on inflammatory mediators and nuclear factor kappa B expression in mouse peritoneal macrophages

The effects of tocotrienol-rich fraction (TRF), α-tocopherol (T) and α-tocopheryl acetate (TA) on lipopolysaccharide (LPS)-induced inflammatory responses in mouse peritoneal macrophages were examined. Results showed that at 5–30 μg/ml, all test compounds plus 1 μg/ml LPS exhibited no cytotoxic effects on macrophage cells. Compared with T and TA, TRF showed the strongest anti-inflammatory activity as demonstrated by its potency in inhibiting the LPS-induced nitric oxide (NO), prostaglandin E₂ (PGE₂), and proinflammatory cytokine (TNF-α, IFN-γ, IL-1β and IL-6) production. At 10 μg/ml, it significantly blocked the LPS induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression, but has no effect on cyclooxygenase-1 (COX-1). Furthermore, TRF also showed a greater inhibition on the nuclear factor kappa B (NF-κB) expression than T and TA. These results suggest that TRF could be a better agent than T and TA for use in the prevention of chronic inflammatory diseases.     

Antioxidative and anti-inflammatory properties of Citrus sulcata extracts

Citrus sulcata was subjected to ultrasound, high-pressure, and Soxhlet extractions. The antioxidant and anti-inflammatory activities of the extracts were evaluated qualitatively and quantitatively. The antioxidant content of the peel extract was twice that of the fruit extract. The quantitative analysis showed that the narirutin and hesperidin contents in the peel extracts were 8.8 and 7.5 mg/100 g, respectively. These extracts had a total phenolic content of 112.22 ± 2.89 gallic acid equivalent (GAE) mg/100 g, a total flavonoid content of 54.09 ± 1.01 rutin equivalent (RE) mg/100 g, DPPH radical scavenging activity of 46%, and antioxidant activity of 213.25 ± 2.82 μM of Trolox equivalents (TEAC). C. sulcata extracts could prevent hydrogen peroxide (H₂O₂)-induced oxidative stress, reduce expression of the inflammatory markers nuclear Factor kappa B (NFκB) and phosphorylated IκBα (p-IκBα) in A549 human lung carcinoma cells, and inhibit Lipopolysaccharide (LPS)-induced THP-1 monocyte differentiation to an extent of 85%.     

沙葱总黄酮水洗组分的体外抗炎活性

本实验在体外应用脂多糖（lipopolysaccharides,LPS）诱导小鼠腹腔巨噬细胞建立炎症模型的基础上,探讨沙葱总黄酮水洗组分的体外抗炎活性。应用CCK-8法检测0、50、100、200、400、800 μg/mL沙葱总黄酮水洗组分对小鼠腹腔巨噬细胞增殖活力的影响;将细胞分为空白对照组、LPS应激模型组及不同质量浓度沙葱总黄酮水洗组分预处理组,采用Griess法及酶联免疫吸附测定法分别测定各处理组细胞上清液中NO浓度和肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白细胞介素（interleukin,IL）-1β、IL-6、IL-10的质量浓度,反转录实时荧光定量聚合酶链式反应法检测各处理组细胞中诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS）、TNF-α、IL-1β、IL-6、IL-10、髓样分化蛋白（myeloid differential protein,MyD）88、核因子 κB（nuclear factor κB,NF-κB）mRNA的表达水平。结果显示：与对照组相比,沙葱总黄酮水洗组分在50～800 μg/mL时对小鼠腹腔巨噬细胞无明显细胞毒性作用。与LPS应激模型组相比,沙葱总黄酮水洗组分能够极显著抑制促炎介质NO、TNF-α、IL-1β、IL-6质量浓度及其mRNA的表达（P＜0.01或P＜0.001）;高度显著提高抗炎细胞因子IL-10质量浓度及其mRNA的表达（P＜0.001）,且呈剂量依赖效应;极显著降低MyD88、NF-κB mRNA的表达水平（P＜0.01或P＜0.001）。由此得出,沙葱总黄酮水洗组分对LPS诱导的小鼠腹腔巨噬细胞具有抗炎作用,其抗炎活性可能是通过抑制促炎性介质NO、TNF-α、IL-1β、IL-6的分泌并提高抗炎性细胞因子IL-10的质量浓度实现的,其作用机制可能与NF-κB信号通路有关。     

Toona sinensis and its major bioactive compound gallic acid inhibit LPS-induced inflammation in nuclear factor-κB transgenic mice as evaluated by in vivo bioluminescence imaging

In the present study, we investigated the anti-inflammatory effects of a nutritious vegetable Toona sinensis (leaf extracts, TS) and its major bioactive compound gallic acid (GA) by analysing LPS-induced NF-κB activation in transgenic mice, using bioluminescence imaging. Mice were challenged intraperitoneally with LPS (1 mg/kg) and treated orally with TS or GA (100 or 5 mg/kg, respectively). In vivo and ex vivo imaging showed that LPS increased NF-κB luminescence in the abdominal region, which was significantly inhibited by TS or GA. Immunohistochemical and ELISA analyses confirmed that TS and GA inhibited LPS-induced NF-κB, interleukin-1β, and tumour necrosis factor-α expression. Microarray analysis revealed that biological pathways associated with metabolism and the immune responses were affected by TS or GA. Particularly, LPS-induced thioredoxin-like 4B (TXNL4B) 2 expression in the small intestine, and TXNL4B, iNOS, and COX-2 expression in RAW 264.7 cells were significantly inhibited by TS or GA. Thus, the anti-inflammatory potential of TS was mediated by the downregulation of NF-κB pathway.     

Suppressive effects of extracts from the aerial part of Coriandrum sativum L. on LPS‐induced inflammatory responses in murine RAW 264.7 macrophages

BACKGROUND: Coriandrum sativum is used not only as a spice to aid flavour and taste values in food, but also as a folk medicine in many countries. Since little is known about the anti‐inflammatory ability of the aerial parts (stem and leaf) of C. sativum, the present study investigated the effect of aerial parts of C. sativum on lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophages. We further explored the molecular mechanism underlying these pharmacological properties of C. sativum. RESULTS: Ethanolic extracts from both stem and leaf of C. sativum (CSEE) significantly decreased LPS‐induced nitric oxide and prostaglandin E₂ production as well as inducible nitric oxide synthase, cyclooxygenase‐2, and pro‐interleukin‐1β expression. Moreover, LPS‐induced IκB‐α phosphorylation and nuclear p65 protein expression as well as nuclear factor‐κB (NF‐κB) nuclear protein–DNA binding affinity and reporter gene activity were dramatically inhibited by aerial parts of CSEE. Exogenous addition of CSEE stem and leaf significantly reduced LPS‐induced expression of phosphorylated mitogen‐activated protein kinases (MAPKs). CONCLUSION: Our data demonstrated that aerial parts of CSEE have a strong anti‐inflammatory property which inhibits pro‐inflammatory mediator expression by suppressing NF‐κB activation and MAPK signal transduction pathway in LPS‐induced macrophages. Copyright © 2010 Society of Chemical Industry     

Protective effect of dried safflower petal aqueous extract and its main constituent,carthamus yellow,against lipopolysaccharide‐induced inflammation in RAW264.7 macrophages

BACKGROUND: Safflower, whose botanic name is Carthamus tinctorius L., is a member of the family Compositae or Asteraceae. Carthamus yellow (CY) is the main constituent of safflower and is composed of safflomin A and safflomin B. Dried safflower petals are used in folk medicine and have been shown to invigorate blood circulation, break up blood stasis, and promote menstruation. In addition, dried safflower petals contain yellow dyes that are used to color food and cosmetics. In this study, we investigated the effects of dried safflower petals aqueous extracts (SFA) and CY on lipopolysaccharide (LPS)‐induced inflammation using RAW264.7 macrophages. RESULTS: Our data showed that treatment with SFA (1–1000 µg mL^?1) and CY (1–2000 µg mL^?1) does not cause cytotoxicity in cells. SFA and CY inhibited LPS‐stimulated nitric oxide (NO), prostaglandin E₂ (PGE₂), and interleukin 1β (IL‐1β) release, through attenuation of inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) protein expression. Further, SFA and CY suppressed the LPS‐induced phosphorylation of nuclear factor‐κB, which was associated with the inhibition of IκB‐α degradation. CONCLUSION: These results suggest that SFA and CY provide an anti‐inflammatory response through inhibiting the production of NO and PGE₂ by the downregulation of iNOS and COX‐2 gene expression. Thus safflower petals have the potential to provide a therapeutic approach to inflammation‐associated disorders. Copyright © 2010 Society of Chemical Industry     

Japanese bog orchid (Eupatorium japonicum) extract suppresses expression of inducible nitric oxide synthase and cyclooxygenase-2 induced by toll-like receptor agonists

Toll-like receptors (TLR) play an important role in the recognition of many pathogen-associated molecular patterns and the induction of innate immunity. Dysregulated activation of TLR signaling pathways is associated with certain inflammatory diseases. Japanese bog orchid (Eupatorium japonicum), which belongs to a family of Asteraceae plants, is consumed as a tea. The present study investigated the effect of the ethanol extracts of flowers of Japanese bog orchid (EJE) on nuclear factor (NF)-κB activation and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) by TLR agonists in murine macrophages. EJE suppressed NF-κB activation and iNOS and COX-2 expression induced by lipopolysaccharide (TLR4 agonist), polyriboinosinic polyribocytidylic acid (TLR3), and macrophage-activating 2 kDa lipopeptide (TLR2 and TLR6). These results suggest that EJE can regulate TLR signaling pathways and indicated its potential as a potent anti-inflammatory drug.