Eighteen structurally diversified sesquiterpenes isolated from Pogostemon cablin and their inhibitory effects on nitric oxide production.

Five new sesquiterpenes, namely, guaianes A-E (1-5), including one novel carbon skeleton guaiane-type sesquiterpene derivatives (1), together with thirteen known compounds (6-18), were isolated from the aerial parts of Pogostemon cablin (Blanco.) Benth. Their chemical structures were mainly established through the relative spectroscopic data, while the absolute configurations of compounds 1-5 were elucidated on the base of single-crystal X-ray diffraction analysis and electronic circular dichroism (ECD) calculations. All compounds were tested for their inhibiting effects on NO production in LPS-stimulated BV2 microglia cells as well as the cell viabilities. The results showed that compounds 2-16 and 18 possessed moderately anti-inflammatory activities at a concentration of 50 µM.

Endothelial Nitric Oxide Mediates the Anti-Atherosclerotic Action of Torenia concolor Lindley var. Formosama Yamazaki.

Torenia concolor Lindley var. formosama Yamazaki ethanolic extract (TCEE) is reported to have anti-inflammatory and anti-obesity properties. However, the effects of TCEE and its underlying mechanisms in the activation of endothelial nitric oxide synthase (eNOS) have not yet been investigated. Increasing the endothelium-derived nitric oxide (NO) production has been known to be beneficial against the development of cardiovascular diseases. In this study, we investigated the effect of TCEE on eNOS activation and NO-related endothelial function and inflammation by using an in vitro system. In endothelial cells (ECs), TCEE increased NO production in a concentration-dependent manner without affecting the expression of eNOS. In addition, TCEE increased the phosphorylation of eNOS at serine 635 residue (Ser635) and Ser1179, Akt at Ser473, calmodulin kinase II (CaMKII) at threonine residue 286 (Thr286), and AMP-activated protein kinase (AMPK) at Thr172. Moreover, TCEE-induced NO production, and EC proliferation, migration, and tube formation were diminished by pretreatment with LY294002 (an Akt inhibitor), KN62 (a CaMKII inhibitor), and compound C (an AMPK inhibitor). Additionally, TCEE attenuated the tumor necrosis factor-α-induced inflammatory response as evidenced by the expression of adhesion molecules in ECs and monocyte adhesion onto ECs. These inflammatory effects of TCEE were abolished by L-NG-nitroarginine methyl ester (an NOS inhibitor). Moreover, chronic treatment with TCEE attenuated hyperlipidemia, systemic and aortic inflammatory response, and the atherosclerotic lesions in apolipoprotein E-deficient mice. Collectively, our findings suggest that TCEE may confer protection from atherosclerosis by preventing endothelial dysfunction.

Harpagophytum procumbens Extract Ameliorates Allodynia and Modulates Oxidative and Antioxidant Stress Pathways in a Rat Model of Spinal Cord Injury.

Spinal cord injury (SCI) is a deliberating disorder with impairments in locomotor deficits and incapacitating sensory abnormalities. Harpagophytum procumbens (Hp) is a botanical widely used for treating inflammation and pain related to various inflammatory and musculoskeletal conditions. Using a modified rodent contusion model of SCI, we explored the effects of this botanical on locomotor function and responses to mechanical stimuli, and examined possible neurochemical changes associated with SCI-induced allodynia. Following spinal cord contusion at T10 level, Hp (300 mg/kg, p.o.) or vehicle (water) was administered daily starting 24 h post-surgery, and behavioral measurements made every-other day until sacrifice (Day 21). Hp treatment markedly ameliorated the contusion-induced decrease in locomotor function and increased sensitivity to mechanical stimuli. Determination of Iba1 expression in spinal cord tissues indicated microglial infiltration starting 3 days post-injury. SCI results in increased levels of 4-hydroxynonenal, an oxidative stress product and proalgesic, which was diminished at 7 days by treatment with Hp. SCI also enhanced antioxidant heme oxygenase-1 (HO-1) expression. Concurrent studies of cultured murine BV-2 microglial cells revealed that Hp suppressed oxidative/nitrosative stress and inflammatory responses, including production of nitric oxide and reactive oxygen species, phosphorylation of cytosolic phospholipases A2, and upregulation of the antioxidative stress pathway involving the nuclear factor erythroid 2-related factor 2 and HO-1. These results support the use of Hp for management of allodynia by providing resilience against the neuroinflammation and pain associated with SCI and other neuropathological conditions.

Phenolic compounds from the aerial parts of Clematis viticella L. and their in vitro anti-inflammatory activities.

Phytochemical investigations on the EtOH extract of Clematis viticella led to the isolation of six flavonoid glycosides, isoorientin (1), isoorientin 3'-O-methyl ether (2), quercetin 7-O-α-L-rhamnopyranoside (3), quercetin 3,7-di-O-α-L-rhamnopyranoside (4), manghaslin (5) and chrysoeriol 7-O-ß-D-glucopyranoside (6), one phenylethanol derivative, hydroxytyrosol (7), along with three phenolic acids, caffeic acid (8), (E)-p-coumaric acid (9) and p-hydroxybenzoic acid (10). The structures of the isolates were elucidated on the basis of NMR and HR-MS data. All compounds were isolated from C. viticella for the first time. Compounds 7 and 8 showed significant anti-inflammatory activity at 100 µM by reducing the release of NO in LPS-stimulated macrophages comparable to positive control indomethacin. Compounds 3 and 7 exhibited anti-inflammatory activity through lowering the levels of TNF-α while 1, 3 and 5 decreased the levels of neopterin better than the positive controls.

Isolation and immunosuppressive effects of 6″-O-acetylginsenoside Rb1 extracted from North American ginseng.

Extraction of medicinally active components from natural health products has become an emerging source for drug discovery. Of particular interest for this work was the finding and testing of a new ginsenoside from North American ginseng (Panax quinquefolius). In the present study, a large amount of 6″-O-acetylginsenoside Rb1, compound 7, was found using ultrasonic extraction of North American ginseng with DMSO aqueous solution. This new ginsenoside was well identified with MS, FTIR, and 1D (1H and 13C) and 2D (gCOSY, gHSQC, and gHMBC) NMR. Subsequent bioassay experiments confirmed that compound 7 demonstrated an additional immunosuppressive activity towards inhibiting the production of nitric oxide and tumor necrosis factor alpha in lipopolysaccharide-induced macrophage cells in a dose-dependent manner using murine macrophages. This new ginsenoside is encouraging for the further exploration and development of novel drugs.

Protective effects of shengmai san and its three fractions on cerebral ischemia-reperfusion injury.

AIM: To investigate the antioxidant and anti-inflammatory effects of Shengmai San (SMS) and its ethyl acetate extract (SEa), n-butanol extract (SBu), and aqueous extract (SWe), and clarify the material base of SMS and the roles played by its fractions. METHODS: A mouse model of transient forebrain ischemia/reperfusion (I/R) by means of common carotid artery occlusion (CCAO) was used to investigate the effects of SMS and its three fractions. Histopathological damage, blood-brain barrier disruption, and antioxidant and inflammation-related parameters, including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), myeloperoxidase (MPO), nitric oxide (NO), tumor necrosis factor-α (TNF-α) were measured. The chemical constituents of each fraction were identified by LC-MS. RESULTS: Eighteen lignans in SEa, and thirteen steroidal glycosides and ginsenosides in SBu were determined. SMS significantly inhibited I/R induced formation of histological injury and cerebral MPO activity. SMS showed the strongest antioxidant and anti-inflammatory effects against the I/R-caused injuries. SEa showed higher antioxidant activity than the other two fractions and SBu has a slightly stronger inhibition on the productions of NO and TNF-α. CONCLUSION: SMS as a whole had the most effective protection against cerebral I/R-caused injuries compared with its fractions, which inferred that it contains different groups of compounds that contribute together to its protective effect.

Natural analcime zeolite modified with 2,3,5,6-tetra(2-pyridyl)pyrazine for preconcentration and determination of trace amounts of cadmium by flame atomic absorption spectrometry.

An analytical method using natural analcime zeolite modified with tppz (2,3,5,6-tetra-pyridylpyrazine) for preconcentration of cadmium, in a column system, and their sequential determination by flame atomic absorption spectrometry (FAAS), was developed. In this work, cadmium was adsorbed onto natural analcime zeolite modified with tppz and then was recovered by nitric acid. Solutions of cadmium were passed through a glass column packed with 100 mg of the sorbent material, at pH 5.0, and cadmium was eluted with 2.0 M HNO(3) at a flow rate of 2.0 ml min(-1). The relative standard deviation (RSD) for eight replicate determinations at the 2.5 µg of cadmium was ±0.94%. The calibration curve using the preconcentration system was linear from 0.01 to 4 µg ml(-1) in final solution with a correlation coefficient of 0.9993. This method was successfully applied for the determination of cadmium in various samples.

Analgesic and anti-inflammatory activities of the methanol extract of Kalanchoe gracilis (L.) DC stem in mice.

In this study, we evaluated the analgesic effect of the methanol extract of Kalanchoe gracilis (MKGS) stem in animal models by inducing writhing response with acetic acid and conducting formalin test. The anti-inflammatory effect of MKGS was also estimated on mice with lambda-carrageenan induced paw edema model. In order to investigate the anti-inflammatory mechanism of MKGS, we analyzed the activities of glutathione peroxidase (GPx) and glutathione reductase (GRx) in the liver, and the levels of interleukin-1beta (IL-1beta), tumor necrosis factor (TNF-alpha), malondialdehyde (MDA) and nitric oxide (NO) in the edema paw tissue. In the analgesic tests, MKGS (0.5 and 1.0 g/kg) decreased both the acetic acid-induced writhing response and the licking time in the late phase of the formalin test. In the anti-inflammatory test, MKGS (0.1, 0.5 and 1.0 g/kg) decreased paw edema at the third, fourth, fifth and sixth hours after lambda-carrageenan had been administrated. Furthermore, MKGS increased the activities of SOD and GRx in liver tissues and decreased MDA level in the edema paws three hours after lambda-carrageenan was injected. MKGS also affected the levels of IL-1beta, TNF-alpha and NO induced by lambda-carrageenan. All these results suggested that MKGS possessed analgesic and anti-inflammatory effects. The anti-inflammatory mechanism of MKGS might be related to the lowering of MDA level in the edema paw via increasing the activities of superoxide dismutase (SOD) and GRx in the liver, as well as the decreases in the levels of TNF-alpha and NO, and the production of IL-1beta in inflamed tissues.

Perinatal micronutrient supplements ameliorate hypertension and proteinuria in adult fawn-hooded hypertensive rats.

BACKGROUND: In fawn-hooded hypertensive (FHH) rats, a model of hypertension, impaired preglomerular resistance, hyperfiltration, and progressive renal injury, we recently observed that supporting perinatal nitric oxide (NO) availability with the NO donor molsidomine persistently reduced blood pressure (BP) and ameliorated renal injury in male and female offspring. However, beneficial effects of perinatal molsidomine treatment were more pronounced in female than in male FHH rats. METHODS: To evaluate whether such protective effects could also be achieved with micronutrients, and whether the gender-dependent differences could be confirmed, we tested perinatal exposure to the micronutrients L-arginine, taurine, vitamin C, and vitamin E (ATCE) in FHH rats. Perinatal micronutrients increased urinary NO metabolite, sodium and potassium excretion only at 4 weeks of age, i.e., at the end of treatment. RESULTS: From 12 weeks onwards, control males had a significantly higher systolic BP (SBP) than females (P < 0.01); however after perinatal micronutrients, this difference was no longer present, indicating a pronounced antihypertensive effect of perinatal micronutrients in males (interaction P < 0.001). Development of proteinuria was attenuated by perinatal micronutrients in males and females. However, only females showed reduced glomerular filtration rate, filtration fraction, and glomerulosclerosis (GS) after perinatal micronutrients. CONCLUSIONS: In sum, perinatal micronutrients that enhance NO availability ameliorated development of hypertension and proteinuria in FHH rats. Antihypertensive effects were more pronounced in male FHH offspring, whereas renal protective effects were more pronounced in female FHH offspring. Mechanisms underlying gender-specific consequences of perinatal micronutrients require further study.

Protective action of a hydroalcoholic extract of a vinifera grape skin on experimental preeclampsia in rats.

OBJECTIVE: This study was designed to determine the protective effects of a vinifera grape skins extract (GSE, 200 mg/kg/day) on the deleterious effect observed in experimental preeclampsia, a condition where reduced nitric oxide production and increase in oxidative stress are present. METHODS: A condition similar to preeclampsia was induced by chronic inhibition of nitric oxide synthesis by L-NAME (60 mg/kg/day, orally) in pregnant rats. Blood pressure (systolic, mean and diastolic) was measured with the tail cuff method on day 20 of pregnant control rats; pregnant rats treated with L-NAME, L-NAME plus GSE or GSE from day 13 to day 20 of pregnancy. Glucose was infused in anesthetized pregnant rats at day 20 and blood glucose and insulin were estimated at time zero, 15, 30, 45 and 60 minutes after beginning of glucose infusion. The number of fetus alive was also estimated at day 20 of pregnancy. In parallel, blood pressure was measured in non-pregnant and in non-pregnant rats treated with L-NAME during 7 days. RESULTS: Increase in arterial pressure, reduction of alive fetus at the end of pregnancy and increase in insulin resistance was observed in pregnant L-NAME rats but not in pregnant L-NAME plus GSE rats or in pregnant GSE rats. Increase in arterial pressure was also observed in non-pregnant L-NAME rats. CONCLUSION: The present study demonstrated a protective effect of GSE in experimental preeclampsia since the deleterious effect induced by L-NAME that is, increased in stillbirth, hypertension and insulin resistance were significantly reduced by oral treatment with the extract. Probably an endothelium-dependent vasodilator effect and an antioxidant action play an important role on the effects of GSE in experimental preeclampsia.

Coronary artery disease, nitric oxide and oxidative stress: the "Yin-Yang" effect--a Chinese concept for a worldwide pandemic.

Prevention of coronary artery disease (CAD) and reduction of its mortality and morbidity remains a major public health challenge throughout the "Western world". Recent evidence supports the concept that the impairment of endothelial function, a hallmark of insulin resistance states, is an upstream event in the pathophysiology of insulin resistance and its main corollaries: atherosclerosis and myocardial infarction. Atherosclerosis is currently thought to be the consequence of a subtle imbalance between pro- and anti-oxidants that produces favourable conditions for lesion progression towards acute thrombotic complications and clinical events. Over the last decade, a remarkable burst of evidence has accumulated, offering the new perspective that bioavailable nitric oxide (NO) plays a pivotal role throughout the CAD-spectrum, from its genesis to the outcome after acute events. Vascular NO is a critical modulator of coronary blood flow by inhibiting smooth muscle contraction and platelet aggregation. It also acts in angiogenesis and cytoprotection. Defective endothelial nitric oxide synthase (eNOS) driven NO synthesis causes development of major cardiovascular risk factors (insulin resistance, arterial hypertension and dyslipidaemia) in mice, and characterises CAD-prone insulin-resistant humans. On the other hand, stimulation of inducible nitric oxide synthase (iNOS) and NO overproduction causes metabolic insulin resistance and characterises atherosclerosis, heart failure and cardiogenic shock in humans, suggesting a "Yin-Yang" effect of NO in the cardiovascular homeostasis. Here, we will present a concise overview of the evidence for this novel concept, providing the conceptual framework for developing a potential therapeutic strategy to prevent and treat CAD.

Synergistic effect of folic acid and vitamin B12 in ameliorating arsenic-induced oxidative damage in pancreatic tissue of rat.

The efficacies of two nutritional factors, folic acid and vitamin B12, were assessed in this study against arsenic-induced islet cellular toxicity. Rats were divided into four groups consisting of five rats in each group: Group A, control; Group B, arsenic-treated; Group C, arsenic+folic acid; and Group D, arsenic+folic acid+vitamin B12. The dose of arsenic, folic acid and vitamin B12, respectively, was 3 mg, 36 microg and 0.63 microg kg(-1) body weight day(-1) for 30 days. Results showed that, compared to control group, there was a significant increase in the levels of nitric oxide (NO), malondialdehyde (MDA) and hydroxyl radical (OH-) formation in the pancreatic tissue of arsenic-treated rats, while the activity of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), and cellular content of antioxidant glutathione (GSH) were low in these animals. The serum level of tumor necrosis factor-alpha (TNF-alpha) and IL-6 was significantly high in these animals. Light microscopic examination showed a marked fall in the number of islet cells. Concomitant administration of either folic acid or folic acid and vitamin B12 with arsenic significantly restored all these parameters. Although folic acid alone could not restore the normal level of TNF-alpha and IL-6, combined folic acid and vitamin B12 could restore it. Folic acid and vitamin B12 combined also could recover islet cell count. These results suggest that folic acid+vitamin B12 are capable of reducing arsenic-induced cellular oxidative and inflammatory toxic changes. Thus, supplement with vitamin B12+folic acid may be predicted as a possible nutritional management strategy against arsenic-induced toxicity.

Generation of organic acids and monosaccharides by hydrolytic and oxidative transformation of food processing residues.

Carbohydrate-rich biomass residues, i.e. sugar beet molasses, whey powder, wine yeast, potato peel sludge, spent hops, malt dust and apple marc, were tested as starting materials for the generation of marketable chemicals, e.g. aliphatic acids, sugar acids and mono-/disaccharides. Residues were oxidized or hydrolyzed under acidic or alkaline conditions applying conventional laboratory digestion methods and microwave assisted techniques. Yields and compositions of the oxidation products differed according to the oxidizing agent used. Main products of oxidation by 30% HNO(3) were acetic, glucaric, oxalic and glycolic acids. Applying H(2)O(2)/CuO in alkaline solution, the organic acid yields were remarkably lower with formic, acetic and threonic acids as main products. Gluconic acid was formed instead of glucaric acid throughout. Reaction of a 10% H(2)O(2) solution with sugar beet molasses generated formic and lactic acids mainly. Na(2)S(2)O(8) solutions were very inefficient at oxidizing the residues. Glucose, arabinose and galactose were formed during acidic hydrolysis of malt dust and apple marc. The glucose content reached 0.35 g per gram of residue. Important advantages of the microwave application were lower reaction times and reduced reagent demands.

Shosaikoto (kampo medicine) protects macrophage function from suppression by hypercholesterolemia.

The feeding of cholesterol-enriched diet for 2 weeks was enough to reduce nitric oxide (NO), prostaglandin E(2) (PGE(2) and interleukin-1 (IL-1) productions in thioglycollate-elicited murine macrophages. Although not showing anti-hypercholesterolemic action against ICR mice, Shosaikoto, a Kampo medicine, partially prevented the reduction of NO and IL-1 productions induced by the feeding of cholesterol-enriched diet, and completely released the reduction of PGE(2) production. These data suggest that the malfunction of macrophage induced by hypercholesterolemia may contribute to early atherogenesis and that Shosaikoto retains macrophage function to prevent the development of atherosclerosis, even though serum cholesterol is markedly increased.