Inhibitory activity of plant extracts on nitric oxide synthesis in LPS-activated macrophages

Nitric oxide (NO) produced in large amounts by inducible nitric oxide synthase (iNOS) is known to be responsible for the vasodilation and hypotension observed in septic shock and inflammation. Inhibitors of iNOS, thus, may be useful candidates for the treatment of inflammatory diseases accompanied by overproduction of NO. We prepared alcoholic extracts of woody plants and screened the inhibitory activity of NO production in lipopolysaccharide (LPS)-activated macrophages after the treatment of these extracts. Among 83 kinds of plant extracts, 23 kinds of extracts showed potent inhibitory activity of NO production above 60% at the concentration of 80 micro g/ml. Some of potent extracts showed dose dependent inhibition of NO production of LPS-activated macrophages at the concentration of 80, 40, 20 micro g/ml. Especially, Artemisia iwayomogi, Machilus thunbergii, Populus davidiana and Populus maximowiczii showed the most potent inhibition (above 70%) at the concentration of 40 micro g/ml. Inhibitory activity of NO production was concentrated to nonpolar solvent fractions (ethyl ether and/or ethyl acetate soluble fractions) of Artemisia iwayomogi, Machilus thunbergii and Morus bombycis. These plants are promising candidates for the study of the activity-guided purification of active compounds and would be useful for the treatment of inflammatory diseases and endotoxemia accompanying overproduction of NO.     

长白山新记录种款冬花提取物体外抗氧化活性研究(英文)

本文分析了长白山新记录种款冬的抗氧化活性。结果表明乙酸乙酯层和正丁醇层款冬提取物有很强的清除自由基和抑制邻苯三酚自氧化活性, 高效液相标定分析 3-咖啡酰奎尼酸, 3, 5-二咖啡酰奎尼酸是主要的抗氧化活性物质, 长白款冬花中 3, 5-二咖啡酰奎尼酸含量达 8.55%, 高于其他省份(0.51%), 这些数据表明长白款冬花是天然的抗氧化植物资源, 具有应用于食品药品产业的潜力。     

高效液相色谱法测定不同产地款冬花的芦丁含量

目的研究巫溪不同产地的款冬所含的芦丁的差异。方法HPLC测定款冬芦丁含量。流动相:甲醇-水(0.025ml/L)磷酸(40∶60);检测波长360nm.结果标准曲线为Y=6934.5X-3365(r=0.9999),芦丁在0.04008~0.4008μg范围呈良好的线性关系;平均回收率为99.40%,RSD为2.14%。结论该方法简便、快捷、准确;巫溪不同产地的款冬花的芦丁含量接近,适合在当地大规模栽培。     

甘肃省款冬花生态适宜性区划研究

目的 通过对甘肃省款冬花的生态适宜性评价,为其资源合理利用提供参考.方法 通过网络资料和实地采集,收集188份款冬花样本,综合55个生态因子,应用地理信息系统和最大熵模型分析款冬花在甘肃省的生态适宜性分布.结果 影响款冬花生态适宜性的主要生态因子分别是4月份降水量、海拔、12月份降水量、等温性、土壤有效水含量等级、土壤...     

超声提取款冬花中绿原酸的工艺研究

目的:优选款冬花中绿原酸的最佳超声提取工艺。方法:以绿原酸为指标,采用单因素考察和正交试验结合的方法,对乙醇浓度、乙醇倍量、提取时间3个因素进行考察,优选提取工艺。结果:乙醇浓度对款冬花中绿原酸的提取率影响最大,其次是提取时间,乙醇倍量影响最小。结论:超声提取款冬花中绿原酸的最佳提取工艺参数为70%乙醇,乙醇倍量为30,提取时间为30 min。     

施肥对款冬生物量分配和产量品质的影响

目的：研究肥料种类和用量对款冬生物量分配、产量和品质的影响。方法：随机区组试验设计,取样测定生物量分配,田间统计产量;分别用HPLC法测定款冬芦丁含量和UV法测定款冬总黄酮含量。结果：在一定肥料施用范围内,肥料种类和用量能明显提高款冬的产量和芦丁、总黄酮含量,并增加生物量向经济器官分配。结论：磷、钾肥能提高款冬花蕾的数量,生产上应适当控制氮肥的用量;氮、磷、钾肥及有机肥在一定施用范围内,能增加款冬花产量和提高款冬花的品质。     

Butanolides from Machilus thunbergii and their inhibitory activity on nitric oxide synthesis in activated macrophages

In activated macrophages the inducible form of nitric oxide synthase (iNOS) generates high amounts of the toxic mediator, nitric oxide (NO), that contributes to the circulatory failure associated with septic shock. Three butanolides were isolated from Machilus thunbergii as active principles which inhibit the production of NO in lipopolysaccharide-activated RAW 264.7 cells, and their structures were identified as litsenolide A2 (1), B1 (2) and B2 (3). They showed dose-dependent inhibition of NO syntheses and the IC(50)s were 3.36, 3.70 and 6.19 micro m, respectively. These new inhibitors of iNOS may have potential in the treatment of endotoxaemia and inflammation accompanied by the overproduction of NO.     

施肥对款冬生物量分配和产量品质的影响

目的：研究肥料种类和用量对款冬生物量分配、产量和品质的影响。方法：随机区组试验设计，取样测定生物量分配，田间统计产量；分别用HPLC法测定款冬芦丁含量和UV法测定款冬总黄酮含量。结果：在一定肥料施用范围内，肥料种类和用量能明显提高款冬的产量和芦丁、总黄酮含量，并增加生物量向经济器官分配。结论：磷、钾肥能提高款冬花蕾的数量，生产上应适当控制氮肥的用量；氮、磷、钾肥及有机肥在一定施用范围内，能增加款冬花产量和提高款冬花的品质。     

吉林省临江市老岭南麓款冬的初步生态研究

目的研究当地款冬的分布及发育状况。方法对款冬的分布、性状、主要遗传参数进行观察研究。结论海拔1 050⁵80 m处有自然分布;花蕾在中秋至晚秋发育迅速,其中款冬酮含量也随之升高;春季葶发育随花的发育逐渐增高;花葶高度受环境影响较大;鳞片叶数随花葶高增高而增加。     

Enhanced expression of inducible nitric oxide synthase by Juzen-taiho-to in LPS-activated RAW264.7 cells, a murine macrophage cell line

We have investigated the effect of Juzen-taiho-to (TJ-48) on inducible NO synthase (iNOS) expression and nitric oxide (NO) production in RAW264.7 cells, a murine macrophage cell line. TJ-48-lipopolysaccharide (LPS) combination induced iNOS mRNA expression earlier, stronger and remained longer that paralleled but with a higher NO production compared to LPS stimulation. TJ-48 itself showed no inducible effect either on NO production or iNOS mRNA expression. This phenomenon could be considered to contribute, at least in part, to the beneficial effects of TJ-48 through the iNOS-mediated activation of biodefense mechanism.     

吉林省临江市老岭南麓款冬自然分布调查

经过不同季节调查观测,吉林省临江市老龄南麓发现的"款冬"植物学性状与《中国植物志》《中国高等植物图鉴》等著作介绍相符,款冬在吉林省临江市老龄南麓有自然分布,分布区间在海拔1050 m⁵80 m范围内。款冬在该区域内均生长良好,甚至在陈煤灰上也生长较好,表现出很强的适应性。秋季根茎上花蕾发育良好,采收加工后与市售款冬花药材无异,应加以开发利用。     

川芎中抑制脂多糖诱导的RAW264.7和BV2细胞系NO生成的新的丁苯酞衍生物——川芎螺内酯

目的研究川芎95%乙醇提取物的化学成分及其抑制脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7和小鼠小胶质细胞BV2一氧化氮(NO)生成的生物学活性。方法采用硅胶、高效液相色谱等柱色谱方法进行分离纯化,通过化合物的谱学数据鉴定其结构。采用LPS离体诱导RAW264.7和BV2细胞系NO生成模型,研究化合物对NO生成的抑制活性。结果从川芎95%乙醇提取物中分离出3个丁苯酞衍生物,分别鉴定为Z-3′,8′,3′a,7′a-四氢-6,3′,7,7′a–二聚藁本内酯-8′-酮(1)、Z,Z′-3.3′a,7.7′a-二聚藁本内酯(2)和川芎螺内酯(3)。对LPS诱导的细胞NO生成抑制作用,在RAW264.7细胞模型,化合物1~3和阳性对照药吲哚美辛的半数抑制浓度(IC_(50))分别为(31.60±2.62)、(21.20±0.61)、(30.12±2.90)、(54.62±7.53)μmol/L;在BV2细胞模型,化合物1~3和阳性对照药姜黄素的IC_(50)分别为(21.99±4.40)、(15.43±1.34)、(12.20±3.40)、(10.58±1.41)μmol/L。结论化合物3为新化合物,命名为川芎螺内酯。生物活性实验结果提示化合物1~3可能具有潜在的抗炎作用。     

Acanthopanax senticosus inhibits nitric oxide production in murine macrophages in vitro and in vivo

Excess nitric oxide (NO) production has been implicated in inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus (A. senticosus) on NO production in murine macrophages in vitro and in vivo. In vitro exposure of RAW264.7 cells to 1, 10, 50, 100, 250, 500 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by lipopolysaccharide (LPS) and interferon gamma (IFN-gamma) in a dose-dependent manner. In vitro exposure of mouse resident peritoneal macrophages to 1, 10, 100 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by LPS and IFN-gamma in a dose-dependent manner. In vivo administration of A. senticosus extract (50, 100 and 200 mg/kg) to KM mice dose-dependently inhibited LPS and IFN-gamma induced production of NO in isolated mouse peritoneal macrophages ex vivo. Exposure to A. senticosus extract had no effect on cell viability and systemic toxicity. The results demonstrated that the stem bark extract of A. senticosus extract inhibits NO production in murine macrophages in vitro and in vivo.     

Maleimide and maleic anhydride derivatives from the mycelia of Antrodia cinnamomea and their nitric oxide inhibitory activities in macrophages

On cultivation of the fungus Antrodia cinnamomea (BCRC 36799) on a medium, the mycelium was extracted and evaluated for nitric oxide (NO) inhibitory activity. Bioactivity-directed fractionation led to the isolation of two new maleimide derivatives, antrocinnamomins A (1) and B (2), and two new maleic anhydride derivatives, antrocinnamomins C (3) and D (4), along with three known compounds, 3-isobutyl-4-[4-(3-methyl-2-butenyloxy)phenyl]furan-2,5-dione (5), 3-isobutyl-4-[4-(3-methyl-2-butenyloxy)phenyl]-1H-pyrrole-2,5-dione (6), and 3-isobutyl-4-[4-(3-methyl-2-butenyloxy)phenyl]-1H-pyrrol-1-ol-2,5-dione (7). Structural elucidation of compounds 1-4 was carried out by spectroscopic data. Compound 1 displayed significant inhibitory effect on nitric oxide (NO) production.     

Down-modulation of nitric oxide production in murine macrophages treated with crude plant extracts from the Brazilian Cerrado

Several plant species from the Cerrado biome in Brazil are popularly used as herbal medicines for its reputed analgesic, anti-acid, anti-microbial, anti-inflammatory and anti-tumoral properties, among others. It has been reported that some plant extracts interfere in the production of nitric oxide (NO), an important inflammatory mediator. In the present study, we investigated the effect of hexanic and ethanolic extracts from three plant species on NO production by LPS/IFN-gamma-activated J774 macrophages based on traditional use. The cytotoxic effect of the crude extracts was determined by the thiazolyl blue test (MTT) to measure cell viability. Serjania lethalis stem extracts and Cupania vernalis leaf extracts significantly inhibited NO production, while extracts from Casearia sylvestris var. lingua were inactive or showed low activity on NO production, or were very cytotoxic. The ethanolic stem bark and leaf extracts of Serjania lethalis and Cupania vernalis, respectively, almost completely inhibited the production of NO by J774 macrophages. It can be concluded that the selected extracts are potential sources of active compounds that might be used as anti-inflammatory agents.     

Acidic polysaccharide isolated from Phellinus linteus enhances through the up-regulation of nitric oxide and tumor necrosis factor-alpha from peritoneal macrophages

Medicinal mushrooms are increasingly used to treat a wide variety of disease processes. Aqueous extract from the fruiting body or mycelia of Phellinus linteus has been reported to produce antitumor and immunomodulatory activities in vivo and in vitro. However, the mechanisms underlying its tumoricidal effects are poorly understood. The tumoricidal activity of peritoneal macrophages (PM) cultured with acidic polysaccharide (PL) isolated from Phellinus linteus against B16 melanoma cells was enhanced in a dose-dependent manner; growth inhibition increased 4-fold with 200 microg/ml of PL. To further characterize the mechanisms of PL, we investigated the effects of PL on phagocytosis and the release of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha) and reactive oxygen intermediates (ROI). To investigate the phagocytosis of PM, the uptake of Dextran (Dex)-FITC between PL-untreated and PL-treated PM was compared. We found some augment in phagocytosis of PL-treated PM compared untreated group. PL stimulated a dose-dependent increase in NO and TNF-alpha, but not in ROI production in PM. We suggested that PL has cytotoxicity against Yac-1 cells through the up-regulation of NO and TNF-alpha production. Also, PL enhanced the expression of costimulatory molecules, CD80 and CD86, and major histocompatibility complex (MHC) molecules II in PM. The ability of PL upon the up-regulation of these surface molecules involved in antigen-presenting processes may, by inference, activate T-cell-mediated immunity against malignant cells in vivo. Taken together, these results suggest that PL act as an effective immunomodulator and enhances the anti-tumoral activity of PM.     

Potent cytotoxic lignans from Justicia procumbens and their effects on nitric oxide and tumor necrosis factor-alpha production in mouse macrophages

A new lignan glycoside, 4-O-alpha-L-arabinopyranosyl-(1' "-->2' ')-beta-D-apiofuranosyldiphyllin (2), named procumbenoside A, and 11 known compounds were isolated from the whole plant of Justicia procumbens. The structure of 2 was established by spectral analysis and chemical methods. The known compounds justicidin A (1), diphyllin (3), and tuberculatin (4) showed potent cytotoxic effects against a number of cancer cells in vitro. Compounds 1 and 4 also strongly enhanced tumor-necrosis factor-alpha (TNF-alpha) generation from mouse macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS).     

Absolute stereostructures of new arborinane-type triterpenoids and inhibitors of nitric oxide production from Rubia yunnanensis

The aqueous acetone extract from the roots of a Chinese herbal medicine, Rubia yunnanensis, showed a potent inhibitory effect on nitric oxide production in lipopolysaccharide-activated macrophages. Five new arborinane-type triterpenes, rubianols-a (1), -b (2), -c (3), -d (4), and -e (5), and a new arborinane-type triterpene glycoside, rubianoside I (6), were isolated from the herbal crude extract together with 10 known compounds. The absolute stereostructures of 1-6 were determined on the basis of chemical and physicochemical evidence, including the application of the modified Mosher's method. The effects of the isolated constituents on nitric oxide production in lipopolysaccharide-activated macrophages were examined, and several triterpenes were found to show inhibitory activity.     

Thai medicinal plants modulate nitric oxide and tumor necrosis factor-alpha in J774.2 mouse macrophages

Centella asiatica (CA) and Rhinacanthus nasutus (RN) are Thai medicinal plants traditionally used to treat a variety of disorders including inflammatory conditions and infections. Nitric oxide (NO) produced from activated macrophages plays a role in both inflammatory and anti-inflammatory processes. This study examined whether CA and RN modulate the production of NO and tumour necrosis factor-alpha (TNF-alpha) by J774.2 mouse macrophages. Expression of the inducible nitric oxide synthase (iNOS) and TNF-alpha genes was also analysed. With CA (water extract) NO production was increased in a dose-dependent manner. An increase also occurred when CA was administered with lipopolysaccharide (LPS), a known macrophage activator. In contrast, an ethanol extract of CA had no effect on NO, and when administered with LPS the extract suppressed production. With RN, neither water nor ethanol extracts alone had an effect on NO, although when the ethanol extract of RN was used in combination with LPS, production was increased. TNF-alpha secretion was correlated with NO production and increases were associated with an elevation in TNF-alpha mRNA. The only effect on iNOS gene expression was an inhibition with the CA ethanol extract in the presence of LPS, consistent with the reduction in NO under these conditions. These studies show that CA and RN extracts can either increase or decrease NO production by macrophages and that these effects are predominantly mediated through an effect on TNF-alpha expression. These data contribute to a better mechanistic understanding of the medicinal properties of these Thai plants.