Sour Cherry Seed Kernel Extract Increases Heme Oxygenase‐1 Expression and Decreases Representation of CD3+ TNF‐α + and CD3 + IL‐8+ Subpopulations in Peripheral Blood Leukocyte Cultures from Type 2 Diabetes Patients

The present study evaluates a hypothesis that sour cherry (Prunus cerasus) seed extracts (SCE) modulate CD3+ T lymphocyte activity in ways predictive of potential for uses of SCE in management of inflammatory diseases. Peripheral blood mononuclear cells (PBMC) from 12 type 2 diabetes (T2DM) patients and eight healthy control subjects were cultured 24 h with 100 ng/ml lipopolysaccharide (LPS) to increase inflammatory signaling and co‐incubated with 0.5–100 µg/ml SCE. Cultures were evaluated by two‐color flow cytometry for percent representation of CD3+ IL8+ and CD3 + TNF‐α + cells which express interleukin‐8 (IL‐8), and tumor necrosis factor‐α, (TNF‐α+) respectively, and by enzyme‐linked immunoassay for lymphocyte‐associated heme oxygenase‐1 (HO‐1, known to be induced by SCE). SCE dosage ranges of 0.5–100 µg/ml in cell cultures significantly suppressed LPS‐increased CD3 + TNF‐α + and CD3 + IL8+ representation from all participants (p < 0.05), with greater pharmacological effect noted in suppression of CD3 + TNF‐α + noted in cells from T2DM patients versus healthy control subjects. These effects correlated with increased HO‐1 expression in SCE‐treated PBMC from all subjects (p < 0.05). Since TNF‐α and IL‐8 are diagnostic/prognostic biomarkers for many inflammatory syndromes, the capacity of SCE to down‐regulate representation of cells that express them suggests potential for therapeutic use of SCE in T2DM and other diseases. Copyright © 2012 John Wiley & Sons, Ltd.     

Protective effect of Morinda citrifolia fruits on β‐amyloid (25–35) induced cognitive dysfunction in mice: An experimental and biochemical study

The neuroprotective effect of an ethyl acetate extract of Morinda citrifolia (Rubiaceae) Linn. fruits (EMC, ethyl acetate extract of Morinda citrifolia) at doses of 200 and 400 mg/kg, p.o. was studied on β‐amyloid (25–35) peptide induced cognitive dysfunction in mice. In the step‐down inhibitory avoidance, EMC exhibited a significant increase in short‐term memory and long‐term memory (p < 0.05). A significant decrease (p < 0.01) in escape latency was noticed in the animals in the water maze. A significant increase (p < 0.01) in alteration of behavior was exhibited upon administration of EMC 200 and 400 mg/kg on the Y maze. Exploratory parameters such as line crossings, head dipping and rearing were increased significantly in EMC treated groups in a dose‐dependent manner (p < 0.05 and p < 0.01). A significant reduction (p < 0.05) in acetyl cholinesterase activity was noticed in the EMC 200 and 400 mg/kg treated groups. The level of monoamine oxidase‐A was decreased by the administration of EMC 200 and 400 mg/kg (p < 0.05 and p < 0.01, respectively). EMC at a dose of 400 mg/kg exhibited a significant increase (p < 0.01) in the levels of serotonin and dopamine. Antioxidant enzymes such as superoxide dismutase, glutathione reductase, glutathione peroxidase and ascorbic acid were decreased significantly in the b‐amyloid peptide injected group, whose levels were restored significantly (p < 0.01) by the administration of EMC (400 mg/kg). Copyright © 2009 John Wiley & Sons, Ltd.     

Antiviral and anticancer activities of 13(E)‐labd‐13‐ene‐8α,15‐diol isolated from Brachyglottis monroi

The antiviral activity of 13(E)‐labd‐13‐ene‐8α,15‐diol (1), isolated from Brachyglottis monroi, was examined against human rhinovirus 2 (HRV2) and 3 (HRV3), and the anticancer activity on human cancer cells (A549 and Hep2). Compound (1) showed strong anti‐HRV2 and HRV3 activity with a 50% inhibitory concentration (IC₅₀) of 2.68 and 0.87 µg/mL, respectively, and a 50% cytotoxicity concentration (CC₅₀) of 59.45 µg/mL. Ribavirin only showed anti‐HRV3 activity with an IC₅₀ of 30.48 µg/mL and a CC₅₀ > 100 µg/mL. The addition of compound (1) to HRV‐infected HeLa cells directly reduced the formation of visible cytopathic effect (CPE) and it directly interacted with HRV particles. Furthermore, A549 and Hep2 cells incubated with 32 µg/mL of compound (1) for 48 h exhibited antilung and antilaryngeal cancer activities, with a viability of less than 50%. These results suggest that compound (1) may be used as a potential antiviral and anticancer agent. Copyright © 2009 John Wiley & Sons, Ltd.     

Chemical Constituents of Euonymus alatus (Thunb.) Sieb. and Their PTP1B and α‐Glucosidase Inhibitory Activities

Phytochemical study on the corks of Euonymus alatus resulted in the isolation of a novel 3‐hydroxycoumarinflavanol (23), along with ten triterpenoids (1–10), ten phenolic derivatives (11–20), and two flavonoid glycosides (21 and 22). Their structures were determined by extensive 1D and 2D‐nuclear magnetic resonance spectroscopic and mass spectrometry data analysis. Furthermore, their inhibitory effects against the protein tyrosine phosphatases 1B (PTP1B) and α‐glucosidase enzyme activity were evaluated. Compounds 6, 7, 9, 15, 19, and 23 were non‐competitive inhibitors, exhibiting most potency with IC₅₀ values ranging from 5.6 ± 0.9 to 18.4 ± 0.3 µm , against PTP1B. Compound 3 (competitive), compounds 5 and 15 (mixed‐competitive) displayed potent inhibition with IC₅₀ values of 15.1 ± 0.7, 23.6 ± 0.6 and 14.8 ± 0.9 µm , respectively. Moreover, compounds 15, 20, and 23 exhibited potent inhibition on α‐glucosidase with IC₅₀ values of 10.5 ± 0.8, 9.5 ± 0.6, and 9.1 ± 0.5 µm , respectively. Thus, these active ingredients may have value as new lead compounds for the development of new antidiabetic agents. Copyright © 2015 John Wiley & Sons, Ltd.     

Down‐regulatory effect of usnic acid on nuclear factor‐κB‐dependent tumor necrosis factor‐α and inducible nitric oxide synthase expression in lipopolysaccharide‐stimulated macrophages RAW 264.7

The purpose of this study was to investigate the molecular mechanisms that are responsible for the antiinflammatory effect of usnic acid (UA). UA is one of the most common and abundant lichen metabolites. The present study examined the effects of UA on the tumor necrosis factor‐α (TNF‐α) and nitric oxide (NO) production induced by lipopolysaccharide (LPS) in RAW264.7 macrophages and the underlying molecular mechanisms. UA decreased the TNF‐α level in LPS‐stimulated RAW264.7 macrophages in dose‐dependent manner, the IC₅₀ value was 12.8 µM. RT‐PCR analysis indicated that it inhibited TNF‐α mRNA expression. Furthermore, it inhibited NO production in LPS‐activated RAW264.7 macrophages, the IC₅₀ value was 4.7 µM. Western blot analysis showed that UA attenuated LPS‐induced synthesis of iNOS protein and nuclear translocation of NF‐κB p65 in the macrophages, in parallel. UA also inhibited LPS‐mediated I‐κBα degradation. Taken together, this suggests that UA has an antiinflammatory effect by inhibiting TNF‐α and iNOS expression, possibly through suppression of nuclear translocation of NF‐κB p65 and I‐κBα degradation. Copyright © 2008 John Wiley & Sons, Ltd.     

Celastrol inhibits growth and metastasis of human gastric cancer cell MKN45 by down‐regulating microRNA‐21

Celastrol could inhibit cancer cell growth in vitro. However, effect(s) of celastrol on gastric cancer is not well studied. Therefore, we investigated the effects of celastrol on human gastric cancer cell line MKN45 and the underlying mechanisms. We found that celastrol inhibited cell proliferation, migration, and invasion and induced cell apoptosis and G2/M cell cycle arrest (p < .05, p < .01, or p < .001). Under celastrol treatment, overexpression of microRNA‐21 (miR‐21) increased cell viability, migration, and invasion and inhibited cell apoptosis compared with negative control (p < .05, p < .01, or p < .001). In addition, the phosphorylation of PTEN was significantly up‐regulated, whereas PI3K, AKT, p65, and IκBα phosphorylation was statistically decreased by celastrol (p < .05 or p < .01) and then further reversed by miR‐21 overexpression (p < .05 or p < .01). On the other side, miR‐21 silence showed contrary results (p < .05) as relative to miR‐21 overexpression. In conclusion, celastrol inhibits proliferation, migration, and invasion and inactivates PTEN/PI3K/AKT and nuclear factor κB signaling pathways in MKN45 cells by down‐regulating miR‐21.     

Medicinal Plants Traditionally Used for Treatment of Obesity and Diabetes Mellitus – Screening for Pancreatic Lipase and α‐Amylase Inhibition

In order to find new pancreatic lipase (PL) and α‐amylase inhibitors from natural sources for the treatment of obesity and related diseases as diabetes mellitus II, 23 medicinal plants with weight‐reducing, serum glucose‐reducing or related potential were investigated. Methanolic and water extracts of the plants were evaluated by using two in vitro test systems. Our findings have shown that the methanolic extract of Hibiscus sabdariffa L. (Malvaceae) showed high inhibitory activities to PL (IC₅₀: 35.8 ± 0.8 µg/mL) and α‐amylase (IC₅₀: 29.3 ± 0.5 µg/mL). Furthermore, the methanolic extract of Tamarindus indica L. (Leguminosae) showed a high anti‐lipase (IC₅₀: 152.0 ± 7.0 µg/mL) and the aqueous extract a high anti‐amylase (IC₅₀: 139.4 ± 9.0 µg/mL) activity. This work provides a priority list of interesting plants for further study with respect to the treatment of obesity and associated diseases. Copyright © 2015 John Wiley & Sons, Ltd.     

Toll‐like receptor 4‐mediated immunoregulation by the aqueous extract of Mori Fructus

The aqueous extract of Mori Fructus (MF) exerts a change of phenotype and a cytoprotective effect in macrophages. The present study was carried out to investigate the immunomodulating activity of MF on the expression of nitric oxide (NO), tumor necrosis factor alpha (TNF‐α), co‐stimulatory molecules and also interferon‐gamma (IFN‐γ) in macrophages and splenocytes. Toll‐like receptor 4 (TLR4) is a promising molecular target for immune‐modulating drugs. It was hypothesized that one possible upstream signaling pathway leading to immunoregulation of MF may be mediated by TLRs. Multiple signaling molecules (NF‐κB, ERK1/2, p38 and JNK) of the TLR4 signaling pathway were also detected. It was found that MF increased NO production and TNF‐α secretion in RAW 264.7 and peritoneal macrophages, co‐stimulatory molecules expression in peritoneal macrophages and IFN‐γ expression in splenocytes. Further studies indicated that MF could significantly induce the phosphorylation of signal molecules of MAPKs and the degradation of IκBα which finally led to the activation and nuclear translocation of nuclear factor‐κB (NF‐κB) for the target gene expression. All those notions disclosed that the aqueous extract MF is a new TLR4 activator, which induces a Th1 immune response as a consequence of induction of cytokines secretion, especially TNF‐α and IFN‐γ. Copyright © 2009 John Wiley & Sons, Ltd.     

Effects of Penta‐O‐galloyl‐β‐D‐glucose on Human Neutrophil Function: significant Down‐Regulation of L‐selectin Expression

Penta‐O‐galloyl‐β‐D‐glucose (PGG) occurrs in high concentrations in medicinal herbs such as Rhus chinensis, Paeonia suffruticosa, Acer truncatum and Terminalia chebula, which demonstrate anti‐inflammatory activity. We investigated the effect of PGG on stimulated and non‐stimulated neutrophils in processes which included reactive oxygen species generation (ROS), metalloproteinase‐9 and interleukin‐8 secretion (IL‐8), β₂ integrin (CD11b) and L‐selectin (CD62L) expression and apoptosis. In concentrations of 5 μM–20 μM, PGG demonstrated statistically significant inhibition of ROS generation, IL‐8 secretion and β₂ integrin expression in stimulated neutrophils. The inhibition of L‐selectin expression by PGG resulted in prevention in neutrophils’ endothelial attachment. The result obtained may explain the anti‐inflammatory activity of this compound and underline the contribution of PGG in the activity of PGG rich plant extracts. Copyright © 2012 John Wiley & Sons, Ltd.     

Bioassay‐guided isolation of apigenin with GABA‐benzodiazepine activity from Tanacetum parthenium

Extracts of Tanacetum parthenium are used in the prophylactic treatment of migraine and have also been used in Danish folk medicine for the treatment of epilepsy. An ethanol extract of T. parthenium showed high affinity for the GABA_A‐benzodiazepine site. An ethanol extract of T. parthenium was fractionated by VLC on silica and preparative C18 HPLC. Each step was monitored with the GABA_A‐benzodiazepine bioassay. The fractionation led to the isolation of apigenin, which may be responsible for CNS‐effects of T. parthenium extracts. Copyright © 2009 John Wiley & Sons, Ltd.     

Antinociceptive properties of 25‐methoxy hispidol A,a triterpinoid isolated from Poncirus trifoliata (Rutaceae) through inhibition of NF‐κB signalling in mice

The 25‐methoxy hispidol A (25‐MHA) is a triterpenoid, isolated from the immature fruit of Poncirus trifoliata (Rutaceae). The pretreatment with 25‐MHA markedly (p < 0.001) attenuated the formalin‐induced biphasic responses as well as acetic acid‐induced writhing responses. The intraperitoneal administration of 25‐MHA significantly attenuated the mechanical hyperalgesia (p < 0.001) and allodynia (p < 0.05). Similarly, 25‐MHA also significantly attenuated (p < 0.001) complete Freund's adjuvant (CFA)‐induced paw edema in mice. The 25‐MHA treatment significantly attenuated the production of nuclear kappa B (NF‐κB) (p65 nuclear subunit). The cytokines are the important mediators of inflammation and pain; however, treatment with 25‐MHA exhibited significant inhibition (p < 0.001) on the mRNA expression levels of various inflammatory mediators. The 25‐MHA administration also significantly enhanced antioxidant enzymes (p < 0.001) and inhibited the oxidative stress markers. The current study indicates that 25‐MHA significantly (p < 0.001) inhibited the nitric oxide (NO) in mice plasma. Similarly, the haematoxylin and eosin (H&E) staining shows that 25‐MHA administration significantly inhibited the inflammatory process in the mice paw tissue compared with the CFA‐treated group. The 25‐MHA treatment did not exhibited any toxicity on the liver, kidney, muscles strength, and motor co‐ordination in mice. The 25‐MHA was coadministered with the various drugs such as tramadol, piroxicam, and gabapentin to observe the synergistic effect.     

The Protective Effect of α‐Hederin,the Active Constituent of Nigella sativa,on Lung Inflammation and Blood Cytokines in Ovalbumin Sensitized Guinea Pigs

In the present study, the preventive effect of two different concentrations of α‐hederin, the active constituent of Nigella sativa, on lung inflammation and blood cytokines in ovalbumin sensitized guinea pigs was examined. Forty eight male adult guinea pigs were divided into control (C), sensitized (S) and sensitized pretreated groups; with thymoquinone (S+TQ), low dose (S+LAH) and high dose of α‐hederin (S+HAH) and inhaled fluticasone propionate (S+FP). The lung histopathology and blood levels of IL‐4, IFN‐γ and IL‐17 were assessed. Compared to sensitized animals, all pathological changes improved significantly in pretreated groups (p < 0.001 to p < 0.05). These improvements in α‐hederin pretreated groups were similar to S+TQ and S+FP groups except cellular infiltration in S+LAH and S+HAH groups which was lower than S+TQ group (p < 0.05). The blood IL‐4 and IL‐17 levels in S+HAH groups showed a significant decrease compared to S group (p < 0.05) which were similar to S+TQ and S+FP groups. The level of IFN‐γ in S+LAH and S+HAH groups increased significantly compared to S group (p < 0.05) which was higher than S+FP group (p < 0.05). Blood IL‐4 in S+HAH group was significantly lower than S+LAH group (p < 0.05). In conclusion, α‐hederin could attenuate the lung inflammation and improve the changes of cytokines like thymoquinone and fluticasone in used dosages. Copyright © 2015 John Wiley & Sons, Ltd.     

1,2,3,6‐tetra‐O‐galloyl‐β‐D‐allopyranose gallotannin isolated,from Euphorbia jolkini,attenuates LPS‐induced nitric oxide production in macrophages

Nitric oxide (NO) is a pleiotropic regulator, critical to numerous biological processes, including vasodilatation and macrophage‐mediated immunity. Macrophages express inducible NO synthase (iNOS) and produce NO after lipopolysaccharide (LPS) stimulation. Gallotannins are water‐soluble polyphenols with wide‐ranging biological activities. Various chemical structures of gallotannins occurring in medicinal and food plants that are used worldwide showed several remarkable biological and pharmacological activities. In the present study, we examined the inhibitory effects of gallotannin 1,2,3,6‐tetra‐O‐galloyl‐β‐D‐allopyranose (GT24) isolated from Euphorbia jolkini on the LPS‐induced NO production and underlying mechanisms of action. GT24 dose‐dependently decreased LPS‐induced NO production and iNOS expression in J774A.1 macrophages. In addition, GT24 inhibited LPS‐induced activation of nuclear factor (NF)‐κB as indicated by inhibition of degradation of I‐κBα, nuclear translocation of NF‐κB, and NF‐κB dependent gene reporter assay. Our results suggest that GT24 possesses an inhibitory effect on the LPS‐induced inflammatory reaction. Copyright © 2010 John Wiley & Sons, Ltd.     

The effect of carvacrol on muscarinic receptors of guinea‐pig tracheal chains

The effects of three concentrations of carvacrol, the constituent of Zataria multiflora Boiss (a monoterpenoid phenol, C₁₀H₁₄O) and 10 nm atropine on muscarinic receptors were tested on: non‐incubated (n = 7), incubated tracheal chains with propranolol and chlorpheniramine (n = 6) and incubated with propranolol (n = 5). The EC₅₀ of all three concentrations of carvacrol in incubated tissues with propranolol and chlorpheniramine was significantly greater than those of incubated tissues with propranolol and non‐incubated trachea (p < 0.05 to p < 0.001). The EC₅₀ of two higher concentrations of carvacrol (0.2 and 0.4 µg/mL) in incubated tissues with propranolol was also significantly greater than those of non‐incubated trachea (p < 0.01 to p < 0.001). The maximum response in the presence of all concentrations of carvacrol in non‐incubated and incubated tissues with propranolol and chlorpheniramine and those of its two higher concentrations (0.2 and 0.4 µg/mL) in incubated tissues with propranolol were lower than saline (p < 0.05 to p < 0.001). There were parallel rightward shifts in the concentration–response curves in the presence of all concentrations of carvacrol in non‐incubated and incubated tissues with propranolol and its lower concentration in incubated tissues with propranolol and chlorpheniramine. These results indicated an inhibitory effect of carvacrol on muscarinic receptors. A β‐adrenoceptor stimulatory effect was also suggested for carvacrol. Copyright © 2010 John Wiley & Sons, Ltd.     

Anti‐allergic activity of principles from the roots and heartwood of caesalpinia sappan on antigen‐induced β‐hexosaminidase release

The dichloromethane extract of the roots and heartwood of Caesalpinia sappan exhibited potent inhibitory activity against β‐hexosaminidase release as marker of degranulation in rat basophilic leukemic (RBL‐2H3) cells, with inhibition of 98.7% and 87.5% at concentration of 100 µg/ml, respectively. These extracts were further separated by chromatographic techniques to give two chalcones and seven homoisoflavones. Among the compounds tested, sappanchalcone (2) possessed the most potent effect against allergic reaction in RBL‐2H3 cells with an inhibitory concentration (IC₅₀) value of 7.6 µM, followed by 3‐deoxysappanchalcone (1, IC₅₀ = 15.3 µM), whereas other compounds showed moderate and mild effects. The results suggested the following structural requirements of chalcones (1 and 2) and homoisoflavones (3‐9) for anti‐allergic activity: (i) chalcone exhibited higher activity than homoisoflavone (ii) vicinal hydroxylation at B‐ring of chalcone conferred higher activity than one hydroxylation; and (iii) for homoisoflavone, the hydroxyl groups at C‐3 and C‐4 positions decreased the activity. This is the first report of C. sappan for anti‐allergic activity. Copyright © 2009 John Wiley & Sons, Ltd.     

Becatamide Found in Houttuynia cordata Suppresses P‐selectin Expression Via Inhibiting COX Enzyme,Not Increasing cAMP in Platelets

Atherosclerosis is a well‐known inflammatory cardiovascular disease. Recent studies suggested potential anti‐atherosclerosis effects of becatamide found in Houttuynia cordata. Therefore, in this study, we investigated potential effect of becatamide (1) and its analogues (enferamide (2), veskamide (3), oretamide (4) and amkamide (5)) on cyclooxygenase (COX)‐1 and ‐2 and the production of cyclic adenosine monophosphate (cAMP), which are critically involved in platelet activation. Among them, becatamide was the most potent compound able to inhibit COX‐1 (IC₅₀ = 0.27 µm ) and ‐2 (IC₅₀ = 0.78 µm ) (p < 0.05). The decreasing order of COX‐1 and ‐2 inhibition activity was becatamide > veskamide > enferamide > oretamide > amkamide. As a result of the inhibition, the production of thromboxane B2 and P‐selectin expression were suppressed by 35% (p < 0.05) and 28% (p < 0.05), respectively, in mouse blood treated with becatamide (0.25 µm ). However, becatamide did not increase intracellular cAMP in platelets. Therefore, the suppression of P‐selectin expression was not blocked by beta 2‐adrenoceptor antagonists, suggesting that the COX inhibition is likely an underlying mechanism for the P‐selectin suppression. In summary, becatamide may be a potent compound to inhibit platelet activation by inhibiting COX enzymes, not by increasing cAMP. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.     

Triterpene acids isolated from Lagerstroemia speciosa leaves as α‐glucosidase inhibitors

The potential antidiabetic activity of ethyl acetate extract of the leaves of Lagerstroemia speciosa (LSL) was investigated by α‐amylase and α‐glucosidase inhibition assay. Six pentacyclic triterpenes (oleanolic acid, arjunolic acid, asiatic acid, maslinic acid, corosolic acid and 23‐hydroxyursolic acid) were isolated from LSL. Their structures were determined by spectroscopic analysis and their α‐glycosidase and α‐amylase inhibitory activities were investigated. They exhibited no or weak inhibitory activity against α‐amylase and middle α‐glucosidase inhibitory activities. Corosolic acid, which shows best bioactivity against α‐glucosidase (IC₅₀ = 3.53 µg/mL), contributes most to the α‐glucosidase inhibitory activity of EtOAc extract. The kinetics of inhibition of corosolic acid was also discussed. Results from this study might provide the scientific evidence for LSL for the treatment of diabetes in traditional medicine. Copyright © 2008 John Wiley & Sons, Ltd.