Anti-inflammatory activity of Chrysanthemum zawadskii var. latilobum leaf extract through haem oxygenase-1 induction

Chrysanthemum zawadskii var. latilobum (CZ) has been widely used as a tea in Korea. In this study, we investigated the involvement of anti-inflammatory haem oxygenase-1 (HO-1) in the inhibitory activity of a CZ leaf extracts on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. At the highest concentration of CZ leaf extract (50 μg/mL), NO production was 10% of that in LPS-stimulated macrophages, whereas iNOS protein expression was 5% of that in LPS-stimulated macrophages. CZ leaf extract also inhibited iNOS mRNA expression in a concentration-dependant manner, and at 50 μg/mL, iNOS mRNA expression was approximately 15% of that in LPS-stimulated cells. The CZ leaf extract induced HO-1 expression in a dose-dependent manner, and blocking HO-1 activity abolished the anti-inflammatory effect of CZ leaf extract. These results suggest that CZ leaf extract has a potent anti-inflammatory activity in RAW264.7 cells through the induction of HO-1.     

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.     

番茄红素对长波紫外线照射大鼠皮肤中丙二醛形成和基质金属蛋白酶表达的抑制作用

为了研究了番茄、野樱莓提取物抗皮肤光老化功效和机制,在UVB辐照HaCaT细胞模型上,研究了番茄、野樱莓提取物对细胞凋亡、活性氧（Reactive oxygen species,ROS）、前列腺素E2（Prostaglandin E₂,PGE₂）的影响。在UVB辐照ICR小鼠模型上,通过苏木精-伊红（hematoxylin-eosin,HE）染色评价番茄野樱莓组合物（LA）抵御皮肤光老化作用。研究皮肤组织中超氧化物歧化酶（superoxide dismutase,SOD）活性、丙二醛（Malondialdehyde,MDA）水平以及促炎因子白细胞介素-1β（Interleukin-1β,IL-1β）、白细胞介素-6（Interleukin-6,IL-6）、环氧化酶-2（Cyclooxygenase-2,COX-2）、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）的表达变化。结果表明,番茄提取物和野樱莓提取物对HaCaT不存在细胞毒性。在HaCaT细胞光老化模型中,与模型组相比,100 μg/mL番茄提取物组和30 μg/mL野樱莓提取物组凋亡细胞数量显著减少（P<0.05）。100 μg/mL野樱莓提取物能够显著降低ROS水平（P<0.01）。100 μg/mL浓度下,野樱莓提取物和番茄提取物组都能够显著降低PGE₂（P<0.05）。与模型组相比,口服LA能够使真皮厚度显著降低（P<0.01）,皮肤组织中SOD活性显著增加（P<0.01）,MDA水平显著降低（P<0.01）,促炎因子IL-6、COX-2、TNF-α显著降低（P<0.05）。番茄、野樱莓提取物组合可以改善UVB诱导的氧化应激和炎症反应,具有抗光老化功效。     

桑葚提取物体外抗炎作用及机制的研究

本文采用脂多糖(LPS)刺激小鼠腹腔巨噬细胞RAW264.7,建立细胞体外的炎症模型,研究桑葚提取物对LPS诱导巨噬细胞RAW264.7分泌功能的影响及其作用机制。实验用1μg/mL LPS刺激RAW264.7细胞,在不同浓度样品的干预下,用MTT法检测不同浓度的样品对RAW264.7细胞的作用;用Griess法检测细胞液中NO的含量;用酶联免疫吸附法(ELISA)检测细胞液中PGE2含量;用免疫印迹法(Western Blot)和RT-PCR法检测桑葚提取物对细胞iNOS和COX-2表达的影响;用HPLC法检测桑葚提取物中白藜芦醇的含量。结果表明桑葚提取物浓度在0.5~2 mg/mL范围内对细胞生长无明显影响;在1~2 mg/mL范围内能有效抑制NO和PGE2的分泌并能有效抑制iNOS和COX-2的表达;桑葚提取物中白藜芦醇的含量为107.44±0.48μg/g。这表明桑葚提取物抑制炎症相关因子表达量,从而减弱促炎症反应,发挥抗炎功效,其抗炎活性可能与桑葚中含有较高的白藜芦醇相关。     

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。     

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。     

4 种迷迭香化合物对脂多糖诱导的RAW264.7细胞氧化应激和炎症的抑制作用

目的：研究4 种迷迭香化合物（鼠尾草酸、鼠尾草酚、迷迭香酚和迷迭香酸）对脂多糖（lipopolysaccharide,LPS）诱导的RAW264.7细胞氧化应激和炎症反应的抑制作用。方法：以LPS诱导RAW264.7细胞构建炎症模型,通过比色法、实时荧光定量聚合酶链式反应、Western blot等方法表征迷迭香化合物对活性氧（reactive oxygen species,ROS）、炎症因子以及相关酶类的影响。结果：上述4 种迷迭香化合物对LPS诱导的细胞内ROS相对含量、丙二醛（malonic dialdehyde,MDA）含量、NO释放量和肿瘤坏死因子α（tumor necrosis factor α,TNF-α）、白细胞介素1β（interleukin 1β,IL-1β）、IL-6 mRNA相对表达量,以及诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS）和环氧合酶2（cyclooxygenase 2,COX-2）蛋白相对表达量的升高有显著抑制作用,对超氧化物歧化酶（superoxide dismutase,SOD）和过氧化氢酶（catalase,CAT）活力有显著保护作用,且具有浓度依赖效应。质量浓度10 μg/mL的鼠尾草酸对LPS诱导的ROS相对含量、NO释放量、TNF-α mRNA相对表达量、MDA含量升高的抑制作用和对SOD和CAT活力的保护作用更好,质量浓度10 μg/mL的迷迭香酚对LPS诱导的IL-1β、IL-6 mRNA相对表达量、iNOS和COX-2蛋白相对表达量升高的抑制作用更好。结论：4 种迷迭香化合物均具有一定的抗氧化和抗炎作用,能缓解炎症反应,其中水溶性迷迭香酸有效质量浓度高于其他3 种。     

Anti-inflammatory effect of the immature peel extract of <Emphasis Type="Italic">Jinkyool</Emphasis> (<Emphasis Type="Italic">Citrus sunki</Emphasis> Hort. ex Tanaka)

The peel of jinkyool (Citrus sunki Hort. ex Tanaka) has been widely used in traditional Asian medicine for treatment of a number of diseases, including indigestion and bronchial asthma. In the present study, we compared the flavonoids content and anti-inflammatory activities of ethanolic peel extracts from both immature and mature fruits. Comparing to the mature peel extract (MPE), the immature peel extract (IPE) contained more abundant flavonoids. IPE more effectively suppressed intracellular reactive oxygen species (ROS) generation (IPE, 67.6±1.2%; MPE, 78.9±2.4% at 300 μg/mL of control), nitric oxide (NO) production, and inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression, as well as interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) mRNA expression in lipopolysaccharide-stimulated RAW 264.7 cells. Additionally, IPE reduced c-Jun NH₂-terminal kinase (JNK) and p38 mitogen-activated kinase (p38) activation, suggesting that IPE may exert an anti-inflammatory effect by suppressing pro-inflammatory cytokines that suppress activation of mitogen-activated protein kinases (MAPKs). Taken together, these results indicate that IPE has potential for use as an anti-inflammatory agent.     

Native Australian fruit polyphenols inhibit COX-2 and iNOS expression in LPS-activated murine macrophages

Purified polyphenolic-rich extracts from four native Australian fruits, Illawarra Plum (Podocarpus elatus Endl., Podocarpaceae), Kakadu Plum (Terminalia ferdinandiana Exell, Combretaceae), Muntries (Kunzea pomifera F. Muell., Myrtaceae) and Native Currant (Acrotriche depressa R.Br., Epacridaceae), were screened for their ability to modulate anti-inflammatory activity in LPS-activated murine macrophages (RAW 264.7). The Kakadu Plum extract inhibited the expression of inducible nitric oxide synthase (iNOS), and importantly was the only fruit in this study that displayed differential inhibition of the expression of cyclooxygenase (COX)-2 but not COX-1. Illawarra Plum (COX-2 and iNOS) and Native Currant (iNOS only) also inhibited inflammatory enzymes, while Muntries exhibited none of these activities under the same conditions. All evaluated extracts inhibited the production of prostaglandin E₂ and nitric oxide, suggesting the involvement of alternative pathways in their regulation for the Muntries extract. Further molecular investigations, showed that Kakadu Plum inhibited the NF-??B pathway, but not the p44/42 mitogen activated protein kinase (MAPK) pathway. Collectively, these results demonstrate potential anti-inflammatory activities of native Australian fruits, in particular Kakadu Plum, in LPS-activated murine macrophages, thus confirming the potential biological activities of these fruits.     

海南无刺蜂蜂蜜中多酚类物质成分分析及其抗氧化、抗炎活性评价

以海南无刺蜂蜂蜜及其多酚类提取物为研究对象,分别测定分析无刺蜂蜂蜜的理化成分和多酚物质,研究其抗氧化和抗炎活性。通过液相色谱-串联四极杆飞行时间质谱分析多酚类成分,并采用福林-酚法和硝酸铝比色法测定多酚类提取物总酚酸和总黄酮含量。采用1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基、2,2’-联氮-双-（3-乙基苯并噻唑啉-6-磺酸）二铵盐自由基（2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate) radical,ABTS＋·）清除实验和铁离子还原力实验评价多酚类提取物的体外抗氧化活性,并采用细菌脂多糖（lipopolysaccharides,LPS）诱导的RAW 264.7细胞体外炎症模型,探究多酚类提取物的抗炎活性。结果显示,无刺蜂蜂蜜水分质量分数为（26.3±0.1）%,pH 3.7±0.2,蛋白质含量为（628.2±9.0）mg/kg,淀粉酶值为（19.6±0.2）mL/（g·h）,总糖质量分数为（46.7±6.0）%,包括果糖（21.0±3.7）%、葡萄糖（24.9±2.2）%、蔗糖（0.8±0.1）%;多酚类提取物的总酚酸和总黄酮含量分别为（96.6±0.4）μg CAE/g和（16.1±0.3）μg QE/g;多酚类提取物的DPPH自由基和ABTS＋?清除能力IC50值分别为（435.1±0.4）、（423.0±0.3）μg/mL,铁离子还原力为（0.6±0.02）μmol Trolox/g;在体外抗炎实验中,多酚类提取物能显著抑制由LPS诱导的RAW 264.7细胞NO的释放,并显著抑制促炎症基因iNOS、IL-1β、IL-6和MCP-1的表达,显著增强抗氧化基因HO-1的表达,并呈剂量相关性。综上所述,海南无刺蜂蜂蜜营养成分丰富,富含酚酸和黄酮类物质,且具有很强的抗氧化和抗炎能力,具有良好的开发利用价值。     

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.     

Effect of mung bean ethanol extract on pro-inflammtory cytokines in LPS stimulated macrophages

The anti-inflammatory effect of mung bean ethanol extract on lipopolysaccharide (LPS) stimulated macrophages (J774) was evaluated. The mung bean extract was separated into 5 fractions by normal phase silica gel column chromatography and the mRNA expressions of pro-inflammatory cytokines were examined after incubation with each fraction in LPS stimulated macrophages. All pro-inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-12β, tumor necrosis factor (TNF)-α, and inducible NO synthase (iNOS) were remarkably decreased in the cells treated with 3.7 mg/mL of F3 fraction. The active F3 fraction did not show any cytotoxcity according to propidium iodide staining and no inhibitory effects on J774 cell proliferation were observed by MTT assay. The active fraction contained gallic acid, vitexin, and isovitexin as major components.     

Suppression of LPS-induced inflammatory activities by Rosmarinus officinalis L.

Rosemary (Rosmarinus officinalis L.) has been used in folk medicine to treat headaches, epilepsy, poor circulation, and many other ailments. It was found that rosemary could act as a stimulant and mild analgesic and could reduce inflammation. However, the mechanisms underlying the anti-inflammatory effects of rosemary need more study to be established. Therefore, in this study, the effects of rosemary on the activation of nuclear factor kappa beta (NF-kB) and mitogen-activated protein kinases (MAPKs), the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and the production of nitric oxide (NO), prostaglandin E₂ (PGE₂), and cytokine in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were investigated. A methanol extract of rosemary and its hexane fraction reduced NO generation with an IC₅₀ of 2.75 and 2.83 μg/ml, respectively. Also, the methanol extract and the hexane fraction inhibited LPS-induced MAPKs and NF-kB activation associated with the inhibition of iNOS or COX-2 expression. LPS-induced production of PGE₂ and tumour necrosis factor-alpha (TNF-α) were blocked by rosemary. Rosemary extract and its hexane fraction are important for the prevention of phosphorylation of MAPKs, thereby blocking NF-kB activation, which in turn leads to decreased expression of iNOS and COX-2, thus preventing inflammation.     

Antioxidant and Anti-Inflammatory Properties of Lower-Polymerized Polyphenols in Oolong Tea

This study investigated the antioxidant and anti-inflammatory properties of the fractionated tea extract, a lower-polymerized polyphenol-rich extract ultrafiltrated from oolong tea. The amounts of total polyphenols, flavonoids and condensed tannin were enriched in fractionated tea extract compared to oolong tea. Furthermore, fractionated tea extract was a stronger scavenger of nitric oxide and 2,2-diphenyl-1-picryhydrazyl radicals than oolong tea. When lipopolysaccharide-treated RAW264.7 macrophages were co-incubated with both tea extracts (500 μg/mL) or (-)-epigallocatechin gallate (100 μg/mL) for 16 h, all samples (oolong tea, fractionated tea extract, and (-)-epigallocatechin gallate) suppressed nitric oxide production by 60, 48, and 46%, respectively, and the suppression was due to nitric oxide-scavenging. However, only fractionated tea extract lowered the proportion of phospholipid arachidonic acid and type II-cyclooxygenase expression, thereby decreasing PGE₂ synthesis by 29%. In conclusion, fractionated tea extract was rich in potent antioxidant substances capable of inhibiting the production of pro-inflammatory mediators.