Vascular barrier protective effects of pellitorine in LPS-induced inflammation in vitro and in vivo

Pellitorine (PT), an active amide compound, is well known to possess insecticidal, antibacterial and anticancer properties. In this study, we first investigated the possible barrier protective effects of pellitorine against pro-inflammatory responses induced by lipopolysaccharide (LPS) and the associated signaling pathways in vitro and in vivo. The barrier protective activities of PT were determined by measuring permeability, monocyte adhesion and migration, and activation of pro-inflammatory proteins in LPS-activated human umbilical vein endothelial cells (HUVECs) and in mice. We found that PT inhibited LPS-induced barrier disruption, expression of cell adhesion molecules (CAMs) and adhesion/transendothelial migration of monocytes to human endothelial cells. PT also suppressed LPS-induced hyperpermeability and leukocyte migration in vivo. Further studies revealed that PT suppressed the production of tumor necrosis factor-α (TNF-α) or Interleukin (IL)-6 and activation of nuclear factor-κB (NF-κB) or extracellular regulated kinases (ERK) 1/2 by LPS. Moreover, treatment with PT resulted in reduced LPS-induced lethal endotoxemia. These results suggest that PT protects vascular barrier integrity by inhibiting hyperpermeability, expression of CAMs, and adhesion and migration of leukocytes, thereby endorsing its usefulness as a therapy for vascular inflammatory diseases.     

A new dicoumarin and anticoagulant activity from Viola yedoensis Makino

A new dicoumarin, named as dimeresculetin (1), together with another dicoumarin, euphorbetin (2) and esculetin (3) were isolated from the ethyl acetate extract of the dried whole plants of Viola yedoensis Makino. The structure of 1 was elucidated as 7-hydroxy-6-[(6,7-dihydroxy-2-oxo-2H-1-benzopyran-5-yl)oxy]-2H-1-benzopyran-2-one on the basis of extensive NMR, as well as the other spectral analysis. Compounds 1–3 exhibited anticoagulant activities with respect to activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT).     

Immunopharmacological properties of flavonoids

The flavonoids are a heterogeneous group of ubiquitous plant polyphenols that abound in the human diet and are endowed with several biological activities, including immunomodulating and antioxidant activities. The bioactivity of flavonoids is tightly correlated with their chemical structure and action mechanisms, mostly inhibitory, on enzymatic systems involved in cellular activation. The aim of this study was to evaluate the effects of acetonic extracts from Lunularia cruciata gametophytes and Feijoa sellowiana fruits on human phagocyte functions. A purified native flavonoid, a catechin, and two of its derivatives were also tested. The extract, and its flavonoid activity, was evaluated by analyzing luminol-dependent chemiluminescence emitted by human circulating phagocytes, at rest or activated by phorbol miristate acetate (PMA). The results demonstrate that both the raw extracts and flavonoids significantly inhibit CL emission by leukocytes, especially when these cells are activated by PMA. The antioxidant activity of flavonoids can be increased by changing the chemical structure of the native molecule.     

Anti-thrombotic activity and chemical characterization of steroidal saponins from Dioscorea zingiberensis C.H. Wright

Steroidal saponins have long attracted scientific attention, due to their structural diversity and significant biological activities. Total steroidal saponins (TSS) extracted from the rhizomes of Dioscorea zingiberensis C.H. Wright (DZW) constitute an effective treatment for cardiovascular disease. However, the active constituents contained in DZW rhizomes and their pharmacological properties are not fully understood. The aim of this work is to determine and quantify the active constituents in DZW rhizomes using fingerprint technique, and evaluate its anti-thrombotic activity using inferior vena cava ligation thrombosis rat model and pulmonary thrombosis mice model after being gavaged with TSS for 1 or 2 weeks. In the study, a chemical fingerprint method was firstly established and validated to quantify and standardize TSS from DZW rhizomes including parvifloside, protodeltonin, protodioscin, protogracillin, zingiberensis saponin, deltonin, dioscin and trillin. TSS extracted from DZW rhizomes were showed to have the inhibitions on platelet aggregation (PAG) and thrombosis, and prolong activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT) in a dose-dependent manner in rats. TSS also prolonged the bleeding time and clotting time in a dose-dependent manner in mice. The results indicate that TSS could inhibit thrombosis by both improving the anticoagulation activity and inhibiting PAG action, suggesting that TSS from DZW rhizomes have the potential to reduce the risk of cardiovascular diseases by anti-thrombotic action.     

AM真菌和紫穗槐苗互作早期宿主防御生理指标的响应特征

在温室盆栽条件下研究AM真菌与紫穗槐互作早期宿主防御生理指标的响应特征。结果表明:在AM真菌侵染宿主植物早期阶段,根系保护酶瞬时激活,植保素类黄酮瞬时积累,而且接种根内球囊霉(GI)同接种摩西球囊霉(GM)处理相比保护酶活性和类黄酮含量变化相对较大。接种AM真菌处理MDA含量下降,尤其在紫穗槐出苗后25天,接种GI和GM处理MDA含量分别降低26.6%和25.8%。在保护酶失活的同时,几丁质诱导子降解产物GlcN和可溶性蛋白含量增加,而且产生一些新的蛋白,由此推测:几丁质诱导子的降解和新增蛋白的出现是调节植物防御反应允许建立互惠共生体的可能机制。     

毛竹叶化学成分对叶部主要害虫影响的通径分析

利用对竹叶可溶性糖、蛋白质、单宁和类黄酮含量的测定和毛竹叶部主要害虫种群的系统调查等资料。采用通径分析研究了竹叶4类化学成分对竹刺瘿螨、南京裂爪螨、竹缺爪螨、竹小爪螨、蠕须盾蚧和刚竹毒蛾等害虫的影响。结果表明：除竹刺瘿螨和南京裂爪螨外，竹叶化学成分对其余4种叶部害且：有着显著影响。这种影响主要表现为对各害虫种群的抑制作用；对于竹缺爪螨，直接作用以可溶性糖的影响最大，间接作用以类黄酮通过可溶性糖的影响较大；对于竹小爪螨，直接作用以单宁的影响最大，间接作用中则以类黄酮通过单宁的影响最大；对于蠕须盾蚧，直接作用以类黄酮的影响最大，间接作用以单宁通过类黄酮的影响最大；对于刚竹毒蛾，直接作用以蛋白质的影响最大，间接作用以类黄酮通过蛋白质的影响最大。     

Effect of Spinosad on Detoxifying and Protective Enzymes of Malacosoma neustria testacea

To study the insecticidal activity and toxicity mechanism of spinosad,we assayed bioactivity of spinosad by leaf membrane method and its effects on the activities of detoxifying and protective enzymes in Malacosoma neustria testacea larvae by spectrophotometry.The results showed that spinosad had an extremely high toxicity against the 4th and 5th instar larvae.The glutathione S-transfer (GST) activity in 4th and 5th instar larvae was firstly inhibited,then induced,and finally inhibited,while the mixed-functional oxidase(MFO) activity was inhibited and then enhanced.The induced effect on carboxylesterase(CarE),the induced and inhibited effect on acetylcholinesterase (AchE) and peroxidase(POD),and the complicated effects on superoxide dismutase(SOD) and Catalase(CAT) were determined in 4th instar larvae with spinosad treatment.However,the obvious inhibition effects were found on the activities of CarE,AchE,SOD and POD in the 5th instar larvae whereas the catalase(CAT) activity was inhibited, then increased,and inhibited finally.Therefore, spinosad could effectively disrupt and interfere not only the detoxifying and protective enzymes but also normal physiological metabolism of M.neustria testacea and show extremely high toxicity against this pest.     

Stereospecific antioxidant effects of ginsenoside Rg3 on oxidative stress induced by cyclophosphamide in mice

Ginsenoside Rg3 is one of the active ingredients isolated from Panax ginseng C.A. Meyer. Previous investigation demonstrated that Rg3 was stereospecific in promotion of the immune response. The present study was designed to evaluate Rg3 and its epimers, 20(R)-Rg3 and 20(S)-Rg3, for their effects on oxidative stress induced by cyclophosphamide (Cy) in mice. Forty-eight mice were randomly distributed into 6 groups and intraperitoneally administered saline solution, Cy, 20(R)-Rg3, 20(S)-Rg3, 20(R)-Rg3+Cy or 20(S)-Rg3+Cy. After that, the spleen, thymus and serum were collected to measure the indices of the organs and oxidative parameters. The results showed that Rg3 significantly inhibited Cy-induced oxidative stress in mice by increasing the indices of the spleen and thymus and total antioxidant capacity, elevating the activities of catalase, superoxidase dismutase and lysozyme as well as decreasing the activity of xanthine oxidase and the levels of malondialdehyde and nitric oxide. Rg3 was stereospecific in antioxidant activities as R form exhibited significantly higher antioxidant effects than S form. Therefore, R form should be used when Rg3 is considered to be used as an antioxidant agent.     

核桃楸树皮提取物的化学成分及其抗氧化活性研究

研究了核桃楸树皮的化学成分及其抗氧化活性.采用Sephadex LH-20柱色谱及薄层色谱等方法进行分离,从其70%丙酮提取物乙酸乙酯及水溶性部分中分到11种化合物,经波谱分析及理化性质鉴定化合物分别为:短叶松素(1)、花旗松素(2)、蛇葡萄素(3)、山奈酚(4)、槲皮素(5)、杨梅素(6)、阿福豆苷(7)、紫云英苷(8)、槲皮苷(9)、异槲皮苷(10)、杨梅苷(11).化合物1、3、5、6、8、10为首次从该植物中分得.经DPPH试验,测定了正己烷溶性、二氯甲烷溶性、乙酸乙酯溶性和水溶性部分以及粗提物和分得化合物的抗氧化活性.其中乙酸乙酯和水溶性部分及化合物1～6与对照组相比具有很强的抗氧化活性.     

喷施赤霉素对欧李果实抗氧化能力的影响

为给调控欧李果实品质提供参考,选用'农大6号'和'农大7号'2个欧李品种果实作为试验材料,分别喷施质量浓度为50、100和150mg/L的赤霉素,测定总酚和类黄酮含量及抗氧化指标。结果表明:赤霉素处理抑制了'农大6号'欧李果实总酚和类黄酮的合成;100mg/L的赤霉素处理促进了'农大7号'欧李果实总酚和类黄酮的合成,50和150mg/L的赤霉素处理抑制了'农大7号'欧李果实总酚和类黄酮的合成。赤霉素对'农大6号'欧李果实总酚和类黄酮的抑制作用要大于'农大7号'。3个抗氧化指标(ABTS、FRAP和DPPH)中,'农大6号'欧李果实的ABTS受赤霉素作用的影响最小,'农大7号'是DPPH受赤霉素作用的影响最小。     

Antifeedant constituents from Fagara macrophylla.

Analysis of the polar fractions of an EtOH extract obtained from the bark of the African medicinal plant Fagara macrophylla led to the isolation and identification of the alkaloids oblongine (6), tembetarine (7) and magnoflorine (8) and the flavonoid hesperidin (9). These compounds, together with other metabolites (1--5) previously isolated from F. macrophylla, were tested for antifeedant activity in a binary-choice bioassay. The acridone alkaloid xanthoxoline (4) was found to have a potent antifeedant activity against larvae of both Spodoptera frugiperda and S. littoralis. 1-Hydroxy-3-methoxy-N-methyl-acridone (2), arborinine (3), tembetarine (7) and magnoflorine (8) were antifeedant against S. frugiperda.     

The vascular protective properties of kinsenoside isolated from Anoectochilus roxburghii under high glucose condition

Anoectochilus roxburghii is a traditional Chinese herb used for the treatment of diabetes and some other diseases. The vascular protective effect of its major active ingredient, kinsenoside, in high glucose conditions was investigated in in vivo and in vitro experiments. In in vivo tests, kinsenoside (50 and 100 mg/kg) efficiently lowered blood glucose and cholesterol levels and it enhanced the oxidation resistance of diabetic mice induced by streptozotocin. In the in vitro assay, kinsenoside (20 and 50 μg/mL) markedly inhibited changes in various biochemical substances (nitric oxide (NO), lactic dehydrogenase (LDH), superoxide dismutase (SOD), and catalase (CAT)) in human umbilical vein endothelial cells (HUVECs) damaged by high glucose (35 mM) and restored vascular endothelial structure by balancing the matrix metalloproteinases–the tissue inhibitors of matrix metalloproteinases (MMP–TIMP) system. The vascular protective effects of kinsenoside were speculated to be attributed to oxidative stress inhibition and the reduction of nuclear factor kappa B (NF-κB) mRNA expression levels in high glucose conditions. Moreover, histological examination, including hematoxylin–eosin (H&E) staining, masson trichrome (Masson) staining, and periodic Schiff-methenamine (PASM) staining, greatly supported the morphological and functional amelioration of diabetes-related changes in mice aortas after kinsenoside (20 and 50 μg/mL) treatment. These results indicated that kinsenoside might be a promising agent for the treatment of diabetic vascular disease.     

菊米总黄酮的超声波提取及其体外抑菌试验

以总黄酮为指标,以水为溶剂采用超声波提取技术,通过单因素试验和正交试验确定菊米的最佳提取方法.同时,采用滤纸片扩散法测定其对7种常见污染菌的最小抑菌浓度(MIC).结果表明:超声波提取菊米总黄酮的最佳工艺条件为:温度70℃,料液比1:120,时间40 min.总黄酮得率可达16.96%;提取物对供试菌中的大肠杆菌、枯草杆菌、金黄色葡萄球菌、啤酒酵母、黄曲霉、青霉具有抑菌作用;对大肠杆菌、枯草杆菌、金黄色葡萄球菌、啤酒酵母具有杀菌作用.总体而言对细菌具有较好的抑菌杀菌作用.     

Activities of digestive and detoxification enzymes in multiple generations of beet armyworm, <Emphasis Type="Italic">Spodoptera exigua</Emphasis> (Hübner), in response to transgenic Bt cotton

To reveal the multi-generation effects of transgenic Bt cotton on the metabolism of secondary target herbivores, the activities of digestive and detoxification enzymes in beet armyworm, Spodoptera exigua (Hübner), fed with transgenic Bt (cv. GK-12) vs. non-Bt cotton (cv. Simian-3) for three successive generations were examined. Significantly lower activities of lipase, carboxylesterase (CarE) and acetylcholinesterase (AChE), and higher activities of trypsin and total superoxide dismutase (T-SOD) were observed in Bt-fed S. exigua than in non-Bt fed individuals during the three generations. But the activity of amylase did not differ between the Bt-fed and non-Bt fed S. exigua. Among the three generations of Bt-fed S. exigua, the activities of lipase and trypsin decreased significantly in the second or third generation than in the first generation. But those of CarE and AChE were increased significantly. Among the three generations of non-Bt fed S. exigua, the activity of lipase was significantly higher in the second and third generations than in the first generation. The increasing adaptation and fitness of S. exigua after continuous exposure to Bt cotton was supposed to be associated with the enhanced activities of detoxification enzymes. Measuring multi-generational enzymes activities of S. exigua in response to Bt cotton can provide proofs of its adaptation development, which is suggested to be a mandatory part of risk assessment of transgenic Bt plants on secondary target herbivores.     

Lactones from Ligusticum chuanxiong Hort. reduces atherosclerotic lesions in apoE-deficient mice via inhibiting over expression of NF-kB -dependent adhesion molecules

The present study aims to investigate the anti-atherosclerotic effects of lactones extracted from Ligusticum chuanxiong Hort (LLC) in apoE-deficient mice (ApoE^−/− mice) and proclaim its underlying mechanisms. Expression of endothelial adhesion molecules and NF-κB around the atherosclerotic lesions was detected by immunohistochemistry (IHC). To further validate the mechanism, effect of LLC on the secretion of ICAM-1 and VCAM-1 of human umbilical vein endothelial cells (HUVECs) induced by tumor necrosis factor α (TNF-α) was measured by ELISA. And the activation of NF-κB was detected by western blot. Mice treated with LLC showed significant reduction in lesion sizes of thoracic segments of the aorta (p < 0.01). Meanwhile, LLC treatments lead to decreases of serum TG, TC and LDL-C contents, respectively. LLC also decreased the expression of CD31, intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1 (MCP-1) and nuclear factor-kappa B (NF-κB) in the atherosclerotic plaque. Moreover, LLC at 3.125–25 μg/mL can dose-dependently attenuate the expression of ICAM-1 and VCAM-1 in TNF-α stimulated HUVECs. Western blot result indicated LLC inhibited activation of NF-κB. These results suggested that LLC could ameliorate atherosclerosis in ApoE^−/− mice. The mechanism of action of LLC on anti-atherosclerotic effect may be attributed to the suppression of the production of NF-κB-dependent adhesion molecules.     

Lathyrane diterpenes from Euphorbia prolifera and their inhibitory activities on LPS-induced NO production

A new lathyrane diterpene (1), an unreported spectroscopic data lathyrane diterpenene (2), and two known analaogues (3 and 4) have been isolated from Euphorbia prolifera. Their structures were elucidated as (12E,2S,3S,4R,5R,6S,9S,11S,15R)-3-butyryloxy-5,15-diacetoxy-6,17-epoxylathyra- 12-en-14-one (1), (12E,2S,3S,4R,5R,6S,9S,11S,15R)-3-propionyloxy-5,15-diacetoxy-6,17- epoxylathyra-12-en-14-one (2), (12E,2S,3S,4R,5R,6S,9S,11S,15R)-3-benzoyloxy-5,15-diacetoxy -6,17-epoxylathyra-12-en-14-one (3), and 15-O-acetyl-17-hydroxyjolkinol (4) by spectroscopic methods (IR, ESIMS, HR-ESIMS, NMR, and X-ray crystallography). The inhibitory activities on LPS-induced NO production of these diterpenes were evaluated and compounds 1, 3 and 4 showed inhibitory effects.     

无瓣海桑对乡土红树植物的化感作用

以外来种无瓣海桑为供体,通过室内栽培实验探讨了无瓣海桑各器官水浸液的不同浓度(即0 1、0 2、0 3、0 4、0 5g·mL-1)对我国4个乡土红树植物的化感作用。结果表明:无瓣海桑对各乡土红树植物均存在化感作用,对胚轴萌发的影响大于对幼苗生长的影响;除最低浓度外,不同浓度的各器官水浸液均能明显抑制各乡土红树植物的萌发,且随着浓度增大抑制作用增强;所有水浸液均表现出高浓度下抑制幼苗生长,随着浓度的降低抑制作用减弱,甚至转为促进的规律;各器官的水浸液对各乡土红树植物的萌发和幼苗生长的抑制作用为:果>根>凋落叶;高浓度时,无瓣海桑的各器官水浸液对不同受体红树植物的幼苗生长抑制强度不同。     

Apoplastic and total peroxidase activities in Scots pine needles at subarctic polluted sites

A gradient survey was carried out in order to compare peroxidase activity in Scots pine (Pinus sylvestris) needles in relation to distance from the industrial centre of Monchegorsk, on the Kola Peninsula in north-western Russia. Apoplastic and total peroxidase activity and sulphur (S), nickel (Ni) and copper (Cu) content in the needles of mature trees were measured on seven plots located between 10 and 110 km from the pollution source. Peroxidase activities in both current- and previous-year needles increased towards the smelters and showed a positive correlation with needle S, Cu and Ni concentrations. Total peroxidase activities showed a more obvious relationship to the pollution gradient in winter than in autumn. The element contents in the current year needles averaged 1649 ppm (S), 128 ppm (Ni) and 118 ppm (Cu) close to the smelters, 1212 ppm (S), 37 ppm (Ni) and 67 ppm (Cu) at a distance of 40 km and 831 ppm (S), 7 ppm (Ni) and 1 ppm (Cu) at the most distant sampling plot.This study showed that both the apoplastic and total peroxidase activities responded to heavy metal and sulphur pollution up to 40 km from the smelters in winter, which indicated an increased oxidative stress in this area. The harsh climate conditions and the high pollution levels may have had additive effects. However, as peroxidases are considered a general indicator of stress, it is not possible to evaluate the extent to which single pollutants contribute to this enzyme activity.