Potent Anti-inflammatory Compound Research Articles

Chrysin, an anti-inflammatory molecule, abrogates renal dysfunction in type 2 diabetic rats

Diabetic nepropathy (DN) is considered as the leading cause of end-stage renal disease (ESRD) worldwide, but the current available treatments are limited. Recent experimental evidences support the role of chronic microinflammation in the development of DN. Therefore, the tumor necrosis factor-alpha (TNF-α) pathway has emerged as a new therapeutic target for the treatment of DN. We investigated the nephroprotective effects of chrysin (5, 7-dihydroxyflavone) in a high fat diet/streptozotocin (HFD/STZ)-induced type 2 diabetic Wistar albino rat model. Chrysin is a potent anti-inflammatory compound that is abundantly found in plant extracts, honey and bee propolis. The treatment with chrysin for 16weeks post induction of diabetes significantly abrogated renal dysfunction and oxidative stress. Chrysin treatment considerably reduced renal TNF-α expression and inhibited the nuclear transcription factor-kappa B (NF-кB) activation. Furthermore, chrysin treatment improved renal pathology and suppressed transforming growth factor-beta (TGF-β), fibronectin and collagen-IV protein expressions in renal tissues. Chrysin also significantly reduced the serum levels of pro-inflammatory cytokines, interleukin-1beta (IL-1β) and IL-6. Moreover, there were no appreciable differences in fasting blood glucose and serum insulin levels between the chrysin treated groups compared to the HFD/STZ-treated group. Hence, our results suggest that chrysin prevents the development of DN in HFD/STZ-induced type 2 diabetic rats through anti-inflammatory effects in the kidney by specifically targeting the TNF-α pathway.

Polymorphic Signature of the Anti-inflammatory Activity of 2,2′-{[1,2-Phenylenebis(methylene)]bis(sulfanediyl)}bis(4,6-dimethylnicotinonitrile)

Weak noncovalent interactions are the basic forces in crystal engineering. Polymorphism in flexible molecules is very common, leading to the development of the crystals of same organic compounds with different medicinal and material properties. Crystallization of 2,2′-{[1,2-phenylenebis(methylene)]bis(sulfanediyl)}bis(4,6-dimethylnicotinonitrile) by evaporation at room temperature from ethyl acetate and hexane and from methanol and ethyl acetate gave stable polymorphs 4a and 4b, respectively, while in acetic acid, it gave metastable polymorph 4c. The polymorphic behavior of the compound has been visualized through single-crystal X-ray and Hirshfeld analysis. These polymorphs are tested for anti-inflammatory activity via the complete Freund’s adjuvant-induced rat paw model, and compounds have exhibited moderate activities. Studies of docking in the catalytic site of cyclooxygenase-2 were used to identify potential anti-inflammatory lead compounds. These results suggest that the supramolecular aggregate str...

A randomized double blind, placebo controlled phase 2 trial of BIIL 284 BS (an LTB4 receptor antagonist) for the treatment of lung disease in children and adults with cystic fibrosis

Airway inflammation, mediated in part by LTB4, contributes to lung destruction in patients with cystic fibrosis (CF). LTB(4)-receptor inhibition may reduce airway inflammation. We report the results of a randomized, double-blind, placebo-controlled study of the efficacy and safety of the leukotriene B(4) (LTB(4))-receptor antagonist BIIL 284 BS in CF patients. CF patients aged ≥6 years with mild to moderate lung disease were randomized to oral BIIL 284 BS or placebo once daily for 24 weeks. Co-primary endpoints were change in FEV(1) and incidence of pulmonary exacerbation. After 420 (155 children, 265 adults) of the planned 600 patients were randomized, the trial was terminated after a planned interim analysis revealed a significant increase in pulmonary related serious adverse events (SAEs) in adults receiving BIIL 284 BS. Final analysis revealed SAEs in 36.1% of adults receiving BIIL 284 BS vs. 21.2% receiving placebo (p = 0.007), and in 29.6% of children receiving BIIL 284 BS vs. 22.9% receiving placebo (p = 0.348). In adults, the incidence of protocol-defined pulmonary exacerbation was greater in those receiving BIIL 284 BS than in those receiving placebo (33.1% vs. 18.2% respectively; p = 0.005). In children, the incidence of protocol-defined pulmonary exacerbation was 19.8% in the BIIL 284 BS arm, and 25.7% in the placebo arm (p = 0.38). While the cause of increased SAEs and exacerbations due to BIIL 284 BS is unknown, the outcome of this trial provides a cautionary tale for the administration of potent anti-inflammatory compounds to individuals with chronic infections, as the potential to significantly suppress the inflammatory response may increase the risk of infection-related adverse events.

Amburanins A and B fromAmburana cearensis: Daphnodorin-Type Biflavonoids that Modulate Human Neutrophil Degranulation

). At concentration higher than 25 µg mL -1 , both compounds suppressed nearly 92% of the neutrophil degranulation and 53% of myeloperoxidase activity, thus indicating that they are potential anti-inflammatory lead compounds.

Facile Synthesis, Characterization and Anti-Inflammatory Effect of 3-Aminoindolizine Derivatives

Four 3-aminoindolizines were synthesized from the substituted 2-bromopyridines and propargyl amines with the aim of obtaining potential anti-inflammatory compounds, which were characterized by NMR, IR, ESI-MS and elemental analysis. Their biological activities were evaluated by the bacterial lipopolysaccharide (LPS) stimulation of mouse cells in the RAW264.7 inflammation model. The target compounds4a-4drevealed moderate anti-inflammatory effects with the inhibitions of 45%~61% at 50 μM. The bioactive 3-aminoindolizines might be used for further optimization as potential anti-inflammatory drugs.

Penicillinolide A: A New Anti-Inflammatory Metabolite from the Marine Fungus Penicillium sp. SF-5292

In the course of studies on bioactive metabolites from marine fungi, a new 10-membered lactone, named penicillinolide A (1) was isolated from the organic extract of Penicillium sp. SF-5292 as a potential anti-inflammatory compound. The structure of penicillinolide A (1) was mainly determined by analysis of NMR and MS data and Mosher’s method. Penicillinolide A (1) inhibited the production of NO and PGE2 due to inhibition of the expression of iNOS and COX-2. Penicillinolide A (1) also reduced TNF-α, IL-1β and IL-6 production, and these anti-inflammatory effects were shown to be correlated with the suppression of the phosphorylation and degradation of IκB-α, NF-κB nuclear translocation, and NF-κB DNA binding activity. In addition, using inhibitor tin protoporphyrin (SnPP), a competitive inhibitor of HO activity, it was verified that the inhibitory effects of compound 1 on the production of pro-inflammatory mediators and NF-κB DNA binding activity were partially associated with HO-1 expression through Nrf2 nuclear translocation.

English

The aim of the present study is to investigate the effect of rutin, a potent antioxidant and anti-inflammatory compound on experimentally-induced diabetic neuropathy (DN) in male Wistar rats. In diabetic and normal rats, the pain-related behavior tests were performed before and after rutin (50 and 100 mg/kg/day for 6 weeks) treatments. In serum, fasting glucose, insulin, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) levels were estimated and in sciatic nerve, thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) levels, and superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione-reductase (GR) and glutathione peroxidase (GSH-Px) activities were measured. Diabetic rats developed neuropathy which was apparent from decreased tail-flick and paw-withdrawal latencies and by following decrease in performance on Rota-Rod treadmill. Rutin treatments ameliorated the hyperalgesia, analgesia and improved motor coordination. Streptozotocin (STZ) significantly increased TBARS and decreased GSH levels in sciatic nerve where rutin treatment significantly protected those changes. In diabetic rats, SOD, CAT, GST, GSH-Px and GR activities were significantly inhibited. Rutin treatment significantly ameliorated decrease in antioxidant defense. Present results demonstrate protective effect of rutin in diabetic neuropathy through attenuation of oxidative stress and they suggest that rutin is a potential drug for the prevention of early diabetic-induced neuropathy. Key words: Streptozotocin, rutin, oxidative stress, neuropathy, diabetes.

The Effects of Intestinal LPS Exposure on Inflammatory Responses in a Porcine Enterohepatic Co-culture System

A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 epithelial cells as upper layer, was developed to study the effects of lipopolysaccharides (LPS) on the gene expression profile of pro-inflammatory cytokines (interleukin-8 (IL-8) and tumor necrosis factor-α) and CYP enzymes (CYP1A1, CYP1A2, CYP3A29). The barrier integrity of IPEC-J2 cells was investigated by transepithelial electrical resistance measurements and by fluorescein isothiocyanate-dextran-based test. Basolateral IL-8 production was significantly elevated in LPS-treated IPEC-J2 and primary hepatocyte mono-cultures as well as in the co-culture system, in a dose-independent manner. The LPS-induced changes in the expression of the CYP1A2 and CYP3A29 genes in hepatocyte mono-cultures differed from those in co-culture after LPS treatment on the apical side of the IPEC-J2 cell layer. CYP1A2 was downregulated by the LPS treatment in mono-cultures but upregulated at 10 μg/ml LPS in co-culture; gene expression of CYP3A29 showed no significant LPS-induced change in the hepatocyte mono-culture but was significantly downregulated in co-culture. The newly established co-culture system capable of mimicking enterohepatic interplay in LPS-induced inflammatory responses in vitro can be used in the future for reliable screening of potential anti-inflammatory compounds.

Neuroprotective Effect of Silymarin by Modulation of Endogenous Biomarkers in Streptozotocin Induced Painful Diabetic Neuropathy

Aim of the present study is to investigate the effect of silymarin (SM), a potent antioxidant and anti-inflammatory compound on experimentally-induced Diabetic Neuropathy (DN) in male Wistar rats. Diabetes was induced by single streptozotocin (STZ) injection in rats. Pain-related behavior tests were performed including tail flick, paw-pressure analgesia and Rota-rod performance. Silymarin treatment was started after 21st day of diabetes induction and continued for 6 consecutive weeks. In serum fasting glucose, insulin, tumor necrosis factor-&alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1&beta (IL-1&beta) levels were estimated and in sciatic nerve, thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH), Superoxide Dismutase (SOD), catalase (CAT), glutathione-s-transferase (GST), glutathione-reductase (GR) and glutathione peroxidase (GSH-Px) activities were measured. Diabetic rats developed neuropathy which was apparent from decreased tail-flick latency and paw-withdrawal latency.This was escorted by decreased motor coordination as assessed by performance on Rota-rod treadmill. Treatment with SM ameliorated the hyperalgesia, analgesia and improved motor coordination. STZ significantly increased TBARS and decreased GSH levels in sciatic nerve where silymarin treatment significantly protected those changes. Enzymatic activities such as SOD, CAT, GST, GSH-Px and GR were significantly inhibited in sciatic nerve of diabetic rats. The SM treatment significantly ameliorated decrease in antioxidant defense. Our results clearly demonstrate protective effect of SM is mediated through attenuation of oxidative stress and suggest therapeutic potential of SM in attenuation of diabetic neuropathy.

Discovery of GW870086: a potent anti-inflammatory steroid with a unique pharmacological profile

Glucocorticoids are highly effective therapies for a range of inflammatory diseases. Advances in the understanding of the diverse molecular mechanisms underpinning glucocorticoid action suggest that anti-inflammatory molecules with reduced side effect liabilities can be discovered. Here we set out to explore whether modification of the 17α position of the steroid nucleus could generate molecules with a unique pharmacological profile and to determine whether such molecules would retain anti-inflammatory activity. The pharmacological properties of GW870086 were compared with fluticasone propionate (FP) using a range of cellular and in vivo model systems, including extensive gene expression profiling. GW870086 repressed inflammatory cytokine release from lung epithelial cells in a similar manner to FP but antagonized the effect of dexamethasone on MMTV-driven reporter gene transactivation. GW870086 had a strong effect on the expression of some glucocorticoid-regulated genes (such as PTGS2), while having minimal impact on the expression of other known target genes (such as SGK). GW870086 retained the ability to strengthen tight junctions in epithelial cell culture but, unlike FP, was unable to protect the culture from elastase-mediated damage. In murine models of irritant-induced contact dermatitis and ovalbumin-induced allergic inflammation, GW870086 showed comparable anti-inflammatory efficacy to FP. GW870086 is a potent anti-inflammatory compound with a unique ability to regulate only a subset of those genes that are normally affected by classical glucocorticoids. It has the potential to become a new topical steroid with a different safety profile to existing therapies.

The Antiinflammatory Action and Pharmacokinetics of a Novel Glucosamine-based Di-Peptide Aminosugar

We have previously shown favorable in vitro gut permeability for three novel di-peptide esters of glucosamine (GlcN) likely facilitated by the peptide transporter 1 (PepT1). Herein, we report the development of a novel assay for the determination of bioavailability of the peptide ester of interest, the anti-inflammatory properties of a glycine-valine ester derivative of GlcN (GVG) as well as its pharmacokinetics under healthy and inflammatory conditions. A pre-column derivatization (with 9-fluorenylmethoxycarbonyl) HPLC assay was developed to study bioavailability of GVG, GlcN or cleaved GlcN in the rats that were cannulated in their right jugular vein for blood collection. The compounds of interest were orally administered to both healthy and arthritic rats. Serial blood samples and urine were collected and assayed for the compounds. The stability of the GVG was also tested after incubation with the rat feces. Efficacy of GVG was tested in inflamed rats (injection of 0.2 mL of Mycobacterium butyricum in squalene) following GVG (20 and 30 mg/kg/day GlcN equivalent) or GlcN (20 and 90 mg/kg/day) administration. Arthritis index was calculated at the end of the experiment. The assay was linear (ranged between 0.05-20 μg/mL) and reproducible (intra- and inter-day<10%). Among the tested compounds, only GVG showed a significantly higher plasma concentrations and urinary excretion than GlcN (≈3-fold increase). GVG showed a favorable stability in the rat feces. Adjuvant arthritis was completely prevented with doses greater than 20 mg/kg/day with GVG being 3-fold more potent than GlcN. The examined glycine-valine-GlcN di-peptide aminosugar is a potent anti-inflammatory compound due to its favorable properties to deliver GlcN into the systemic circulation.

In-vitro antioxidant and in-vivo anti-inflammatory activities of aerial parts of Cassia species

Cassia species are native to Southeast Asia and sub-Saharan Africa and are commonly used as food and in a broad variety of medicinal applications. Cassia species are widely used in India, but there are few reports in the literature of studies on its chemical compositions and biological properties. In this study, the antioxidant and anti-inflammatory activities of two plants of Cassia species namely Cassia siamea (Lam.) and Cassia javanica (Linn.) were evaluated and the total phenolic compounds and flavonoid contents were determined.The antioxidant activity of the extracts was measured using scavenging of 2,2′-Diphenyl-1-picrylhydrazyl hydrate (DPPH), bleaching of β-carotene and % inhibition of H2O2. The anti-inflammatory activity was evaluated using carrageenan induced paw edema method on Wistar albino rats. The etahnolic extracts of aerial parts of C. siamea and C. javanica were evaluated for in vivo anti-inflammatory activity against the animal model of female Wistar albino rats. Ethanol extracts showed significant and dose-dependent anti-inflammatory effects. The contents of flavonoids and total phenolic compounds could be correlated with the antioxidant and anti-inflammatory activities observed for C. siamea and C. javanica. Our findings suggest that aerial parts of C. siamea and C. javanica contain potential antioxidant and anti-inflammatory compounds, which could be tested as drug candidates against oxidative and inflammation-related pathological processes in medicinal chemistry studies.

Inhibitor Effect of Antioxidant Flavonoids Quercitin, and Capsaicin in Mast Cell Inflammation

Mast cells are essential not only for allergies but also for innate and acquired immunity, autoimmunity and inflammation, and they are recognized as a new type of immunoregulatory cells capable of producing different cytokines. Natural compounds have long been recognized to possess anti-inflammatory, antioxidant and anticancergenic activity. Quercitin is an inhibitor for mast cells and is a potent antioxidant, cytoprotective and anti-inflammatory compound and has a negative effect on intracellular regulator signal events initiated by FceRI receptor cross-linking and other activating receptors on mast cells. These observations candidate quercitin as a therapeutic compound in association with other therapeutic molecules. Capsaicin is a compound derived from peppers, especially capsicum, and is involved in stimulating circulation aiding digestion and relieving pain. Capsaicin receptor sub type I (VRI) is expressing in neurons and is present in a number of brain nuclei and in non-neuronal tissues, mediating inflammatory response. Capsaicin is involved in migraine, allergic symptoms, arthritis pain and gastric secretion. In this paper we review the biological effects of quercitin and capsaicin.

Design, microwave assisted synthesis and pharmacological activities of substituted pyrimido[2,1-b][1,3]benzothiazole-3-carboxylate derivatives

Background and aimThe current need for the discovery and development of new lead compounds exhibiting optimal antitumor potency as well as anti-inflammatory activity. Due to biological activity of significant number of compounds containing condensed pyrimidine ring system and our interest in MCRs reactions, we wish to report the synthesis of 4H-pyrimido[2,1-b][1,3]benzothiazole-3-carboxylate. MethodA novel one pot multi-component reaction of substituted benzaldehyde, ethyl cyanoacetate, 2-amino benzothiazole to give pyrimido[2,1-b][1,3]benzothiazole-3-carboxylate was described. The operational simplicity, environmental friendly conditions, regioselective formation of target product and high yield in significantly very short reaction time were major benefits. Result and discussionStructural assignments based on spectroscopic data (FT-IR, 1H NMR, mass spectra). Compounds evaluated for in vitro on panel of 60 different human tumor cell lines derived from nine neoplastic cancer types at NCI, and for the in vitro anti-inflammatory activity. Compound ethyl(4R)-2-amino-6,9-dimethyl-4-[4-(dimethylamino)phenyl]-4H-pyrimido[2,1-b][1,3]benzothiazole-3-carboxylate (4i) was found to possess good antiproliferative activity and compound ethyl(4R)-2-amino-6-chloro-4-(3,4,5-trimethoxyphenyl)-4H-pyrimido[2,1-b][1,3]benzothiazole-3-carboxylate (4g) was found to posses potent anti-inflammatory activity and compounds 4a, 4b, 4h also found to be significant anti-inflammatory activity. ConclusionThe obtained results prove the necessity for further investigations to clarify the features underlying the antitumor potential of tested compounds and had potential to design lead for anti-inflammatory activity.

Molecular docking of Diospyrin as a LOX inhibitory compound

Diospyros lotus is traditionally used in various diseases including inflammation. In the current study an effort has been made to identify a bioactive constituent from D. lotus in order to scientifically validate its use in inflammatory disorders. Diospyrin was isolated from D. lotus and exhibited significant lipoxygenase (LOX) inhibitory activity (IC50 value: 31.89±0.14μmol). Molecular docking revealed significant molecular interactions between Diospyrin and LOX showing promising potential for further optimization as a potential anti-inflammatory lead compound.

Lipopolysaccharide (LPS) segmental lung challenge in nonhuman primates – a model of airway inflammation

We developed a reproducible model of neutrophilic inflammation in cynomolgus monkeys (Macaca fascicularis) in order to study potential novel anti-inflammatory compounds intended for respiratory diseases such as COPD and asthma. Using a pediatric bronchoscope, a baseline bronchoalveolar lavage (BAL) was performed on the left lung prior to LPS segmental challenge to the right lung. At 24, 72 and 168 hours post challenge, BAL fluid (BALF) was collected to assess the level of pulmonary inflammation. At both 24 and 72 hours (Hr) post LPS challenge, total BAL cells were significantly elevated over baseline. BAL cell differentials revealed that mean percent neutrophils significantly increased from baseline (1.3 ±1.6%) to (74.0 ± 21.9% at 24 Hr and 26.6 ± 21.0% at 72 Hr) whereas, BAL macrophage numbers had significantly decreased from baseline (94.4 ± 3.1%) to (31.6 ± 21.2 % at 24 Hr and 64.2 ± 20.1% at 72 Hr) post challenge. In addition to increased inflammation, we also observed a significant increase in BAL cell gene expression of MMP-9 and significant increases in sCD14 and IL-8 from concentrated BALF 24 Hr post challenge. Numbers of infiltrating cells and other inflammatory parameters returned to baseline levels by 168 Hr following LPS challenge. In some studies, intramuscular injections of dexamethasone (Dex [1 mg/kg]) were administered 24 Hr prior and just prior to LPS challenge. Dex did not significantly reduce total BAL inflammation or IL-8 protein, but did significantly reduce percent neutrophils and sCD14 protein within the BALF 24 hours post challenge. Dex also significantly increased percent macrophages 24 Hr post challenge. These data show that segmental challenge with LPS produces an acute pulmonary neutrophilic inflammation, which is reduced by pretreatment with Dex. This reproducible, short-term, large animal model may help us to better understand neutrophillic inflammation pathways contributing to lung injury and host defense.

The Effect of Sodium Tanshinone IIA Sulfate and Simvastatin on Elevated Serum Levels of Inflammatory Markers in Patients with Coronary Heart Disease: A Study Protocol for a Randomized Controlled Trial.

Background. Coronary heart disease (CHD) due to atherosclerotic inflammation remains a significant threat to global health despite the success of the lipid-lowering, anti-inflammatory statins. Tanshinone IIA, a potent anti-inflammatory compound derived from Traditional Chinese Medicine (TCM), may be able to supplement statins by further reducing levels of circulating inflammatory markers correlated to cardiovascular risk. Here, we present the protocol of a randomized controlled trial (RCT) that will investigate the synergistic effect of sodium tanshinone IIA sulfate and simvastatin on reducing elevated inflammatory markers in patients with CHD. Participants: Seventy-two inpatients with confirmed CHD, elevated serum high-sensitivity C-reactive protein (Hs-CRP) level, and a TCM diagnosis of blood stasis syndrome will be enrolled and randomized 1 : 1 into the control or experimental group. Intervention. All subjects will receive a standard Western therapy including 20 mg simvastatin orally once per evening. Patients in the experimental group will additionally receive a daily 80 mg dose of sodium tanshinone IIA sulfate intravenously, diluted into 250 mL 0.9% NaCl solution. The treatment period will be 14 days. Outcomes. Primary outcome parameter: serum Hs-CRP level. Secondary outcome parameters: other circulating inflammatory markers (including IL-6, TNFα, VCAM-1, CD40, sCD40L, MCP-1, and MMP-9), improvement in symptoms of angina and blood stasis syndrome, and safety. This trial is registered with ChiCTR-TRC-12002361.

Curcumins-Rich Curry Diet and Pulmonary Function in Asian Older Adults

BackgroundResearch on the effects of dietary nutrients on respiratory health in human populations have not investigated curcumin, a potent anti-oxidant and anti-inflammatory compound present principally in turmeric used in large amounts in Asian curry meals.ObjectivesTo examine the association of curry intake with pulmonary function among smokers and non-smokers.DesignThe frequency of curry intake, respiratory risk factors and spirometry were measured in a population-based study of 2,478 Chinese older adults aged 55 and above in the Singapore Longitudinal Ageing Studies.ResultsCurry intake (at least once monthly) was significantly associated with better FEV1 (b = 0.045±0.018, p = 0.011) and FEV1/FVC (b = 1.14±0.52, p = 0.029) in multivariate analyses that controlled simultaneously for gender, age, height, height-squared, smoking, occupational exposure and asthma/COPD history and other dietary or supplementary intakes. Increasing levels of curry intake (‘never or rarely’, ‘occasional’, ‘often’, ‘very often’) were associated with higher mean adjusted FEV1 (p for linear trend = 0.001) and FEV1/FVC% (p for linear trend = 0.048). Significant effect modifications were observed for FEV1 (curry* smoking interaction, p = 0.028) and FEV1/FVC% (curry*smoking interaction, p = 0.05). There were significantly larger differences in FEV1 and FEV1/FVC% between curry intake and non-curry intake especially among current and past smokers. The mean adjusted FEV1 associated with curry intake was 9.2% higher among current smokers, 10.3% higher among past smokers, and 1.5% higher among non-smokers.ConclusionThe possible role of curcumins in protecting the pulmonary function of smokers should be investigated in further clinical studies.

English

Aralia cordata is known in traditional Korean medicine as a remedy for arthritis. As a part of our program to screen medicinal plants for potential anti-inflammatory compounds, oleanoic acid 28-O-β-D-glycopyranosyl ester (OA) was isolated from the roots of Aralia cordata. So, the immune modulation molecular mechanism of OA isolated from A. cordata was studied.One of the possible mechanisms for its protective activities is by down regulation of the inflammatory responses. Therefore, cells from the human mast cell line (HMC-1) were used to investigate this effect. OA significantly inhibits the way in which phorbol 12-myristate 13-acetate (PMA) plus A23187 induces the production of inflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8 and intracellular Ca2+ levels. In activated HMC-1 cells, phosphorylation of extra-signal response kinase (ERK) 1/2, degradation of IκB, and activation of NF-κB decreased after treatment with OA. OA suppressed the expression of TNF-α, IL-6 and IL-8 through a decrease in the intracellular levels of Ca2+ and ERK 1/2, as well as activation of NF-κB. These results indicated that OA exerted a regulatory effect on inflammatory reactions mediated by mast cells. Key words: Oleanoic acid 28-O-β-D-glycopyranosyl

Bioassay guided isolation and identification of anti-inflammatory and anti-microbial compounds from Urtica dioica L. (Urticaceae) leaves

The present study describes the anti-inflammatory, anti-microbial activity and lipophilic profile with acute toxicological studies of Urtica dioica. Successive extraction of the leaves with organic solvents of increasing polarity and their screening for anti-inflammatory and anti-microbial activity was assessed. Hexane extract showed good anti-inflammatory and anti-microbial activity; hence it was further fractionated using open silica gel column chromatography into 19 sub fractions which were pooled together according to their thin layer chromatography (TLC) profile to give an overall 5 fractions. Among the 5 fractions, fraction-II (FII) at a dose of 200 mg/kg body-weight (bw) exhibits equipotent anti-inflammatory activity (48.83% after 3 h) as that of the standard drug indomethacin (53.48%) in Wistar rats. FII also showed a potent anti-microbial activity against all the tested bacterial strains and its minimum inhibitory concentration (MIC) value was 125, 15.62, 31.25, 250, 31.25, 125 and 7.81 µg/ml against Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae,Pseudomonas aeruginosa, Staphylococcus aureus, Shigella flexneri and Salmonella typhi, respectively which was determined by serial tube dilution method. FII was subjected to gas chromatography-mass spectrometry (GC-MS) analysis in search of potent anti-inflammatory and anti-microbial compound(s). 2,4-Di-t-butylphenol (4.56%), neophytadiene (26.97%), butyl tetradecyl ester (9.53%), dibutyl phthalate (7.45%), bis(2-ethyl hexyl) maleate (8.80%), 1,2-benzenedicarboxylic acid (9.89%) and 2-tert-butyl-4,6-bis(3,5-di-tert-butyl-4-hydroxybenzyl)phenol (3.19%) were the major constituents responsible for both anti-inflammatory and anti-microbial activity of hexane extract of U. dioica. Sub-acute oral toxicity of crude n-hexane extract of U. dioica was carried out in Wistar rats at doses of 250, 500, 1000 and 2000 mg/kg bw to assess the safety index. Hematological parameters from blood and other biochemical parameters from serum confirmed its safety at tested concentrations. Our results corroborate the anti-inflammatory and anti-microbial activity of U. dioica, and could justify its use in folk medicine for the treatment of rheumatic arthritis and other infectious diseases. Key words: Urtica dioica, anti-inflammatory, anti-microbial, minimum inhibitory concentration (MIC), gas chromatography-mass spectrometry (GC-MS), toxicity.