Research progress in drugs targeting 5-lipoxygenase for age-related diseases / 生理学报

With the acceleration of aging society, delaying aging or promoting healthy aging has become a major demand for human health. 5-Lipoxygenase (5-LOX) is a key enzyme catalyzing arachidonic acid into leukotrienes (LTs), which is a potent mediator of the inflammatory response. Previous studies showed that abnormal activation of 5-LOX and overproduction of LTs are closely related to the occurrence and development of aging-related inflammatory diseases. Therefore, inhibiting 5-LOX activation is a possibly potential strategy for treating age-related diseases. In this paper, the latest research progress in 5-LOX activation, 5-LOX in mediating aging-related diseases and its small molecule inhibitors is briefly reviewed to provide scientific theoretical basis and new ideas for the prevention and treatment of aging-related inflammatory diseases.

In vitro pharmacological attributes and metabolite's fingerprinting of Conocarpus lancifolius / Atributos farmacológicos in vitro y huellas dactilares de metabolitos de Conocarpus lancifolius

Search for safe antioxidants and novel nutraceuticals urged to evaluate the antioxidant, anti-acetylcholine esterase and anti-lipoxygenase activity of various leaf extracts of Conocarpus lancifolius. Extraction was optimized from freeze dried plant extracts quenched with liquid nitrogen using water, ethanol, methanol, hexane, ethyl acetate and chloroform. Maximum extract yield, total phenolic contents and total flavonoid contents were obtained in case of ethanolic extraction. The highest 2,2-diphenyl-1-picrylhydrazylradical scavenging in terms of IC50 value of 55.26 µg/mL was observed for ethanolic leaf extract. The acetylcholine esterase and lipoxygenase inhibitory activities (IC50) were also observed for ethanolic extract. These findings for ethanolic extract were statistically significant when compared with other extracts (ρ<0.05). The haemolytic % values indicated that all extracts were associated with very low or negligible toxicity. The epicatechin, isorhamnetin, rutin, scopoleptin, skimmianine, quercetin-3-O-α-rhamnoside, quercetin-3-O-ß-glucoside, cornoside, creatinine, choline, pyruvic acid, α-hydroxybutyric acid, phyllanthin and hypophyllanthin were identified as major functional metabolites in ethanolic leaf extract of C. lancifoliusby 1H-NMR. The identified metabolites were probably responsible for the pharmacological properties of C.lancifolius. The findings may be utilized as pharmacological leads for drug development and food fortification.

Se insta a la búsqueda de antioxidantes seguros y nuevos nutracéuticos para evaluar la actividad antioxidante, anti-acetilcolina esterasa y anti-lipoxigenasa de varios extractos de hojas de Conocarpus lancifolius. La extracción se optimizó a partir de extractos de plantas liofilizados enfriados con nitrógeno líquido usando agua, etanol, metanol, hexano, acetato de etilo y cloroformo. En el caso de extracción etanólica se obtuvo el rendimiento máximo de extracto, el contenido de fenoles totales y el contenido de flavonoides totales. La mayor eliminación de radicales 2,2-difenil-1-picrilhidrazilo en términos de valor de CI50 de 55,26 µg/mL se observó para el extracto de hoja etanólico. También se observaron las actividades inhibidoras de la acetilcolina esterasa y lipoxigenasa (CI50) para el extracto etanólico. Estos hallazgos para el extracto etanólico fueron estadísticamente significativos en comparación con otros extractos (ρ<0.05). Los valores del % hemolítico indicaron que todos los extractos estaban asociados con una toxicidad muy baja o insignificante. Se identificaron la epicatequina, isorhamnetina, rutina, escopoleptina, skimmianina, quercetina-3-O-α-ramnosido, quercetina-3-O-ß-glucósido, cornosido, creatinina, colina, ácido pirúvico, ácido α-hidroxibutírico, filantrina e hipofillantina. como metabolitos funcionales principales en el extracto etanólico de hojas de C. lancifoliuspor 1H-NMR. Los metabolitos identificados probablemente fueron responsables de las propiedades farmacológicas de C. lancifolius. Los hallazgos pueden utilizarse como pistas farmacológicas para el desarrollo de fármacos y la fortificación de alimentos.

Efeito tempo-dose-resposta do montelucaste no processo de reparo alveolar em camundongos 129Sv/Ev / Time-dose-response effect of Montelukast in the process of alveolar repair in 129Sv / Ev mice

Metabólitos do ácido araquidônico são conhecidos por exercerem importante papel nos processos inflamatórios e no metabolismo do tecido ósseo. No entanto, as ações pontuais, especialmente dos leucotrienos derivados da 5-lipoxigenase (5-LO) sobre o processo de reparo ósseo intramembranoso são pouco exploradas. O presente estudo tem como objetivo analisar os efeitos tempo-dose-resposta da droga montelucaste (MTK), potente antagonista dos receptores de cisteinil leucotrienos tipo 1 (CisLT1Rs), no curso do reparo alveolar pós-exodontia em camundongos 129Sv/Ev, bem como nos níveis plasmáticos de marcadores ósseos bioquímicos. Para tanto, foram utilizados 70 camundongos machos jovens divididos em quatro grupos, de acordo com o tratamento: C - Grupo Controle (não tratados); CV - Grupo Controle Veículo, 20 µL de solução fisiológica (SF) 0,9%; MTK2 ­2 mg/kg de MTK e MTK4 ­ 4 mg/kg de MTK. Os animais dos grupos CV, MTK2 e MTK4 foram tratados diariamente por via oral, iniciando 24 horas antes do procedimento cirúrgico, continuando até o final dos períodos experimentais de 7, 14 e 21 dias pós-operatórios. Ao final dos períodos determinados, os animais foram submetidos à eutanásia para coleta de sangue para análise bioquímica dos níveis de cálcio, fosfato, fosfatase ácida resistente ao tartarato (TRAP) total e fosfatase alcalina (FAL), coleta da maxila direita contendo os alvéolos dentários para serem analisados por meio de microtomografia computadorizada (microCT), e análise histopatológica. Os resultados obtidos foram submetidos à testes estatísticos considerando-se nível de confiança de 5%. Observou-se aumento do BV/TV para os animais tratados com MTK em relação aos grupos C e CV, tanto aos 14 dias quanto aos 21 dias, sendo maior no grupo MTK4 aos 14 dias em relação ao grupo MTK2. Do mesmo modo, os animais tratados com MTK em ambas doses apresentaram aumento significativo de Tb.Th em comparação aos grupos C e CV aos 21 dias. Chamou a atenção valores de BV/TV e Tb.Th significativamente reduzidos no grupo CV em comparação ao C, indicando um efeito negativo da manipulação do animal. Na análise histopatológica observou-se reparo ósseo precoce nos animais MTK2 e MTK4 em todos os períodos avaliados, em comparação aos do grupo C, bem como atraso no processo de reparo no grupo CV aos 21 dias. Quanto aos marcadores plasmáticos, observou-se aumento do cálcio no grupo MTK4 em relação ao grupo C aos 7 dias, e aos 21 dias também em relação ao grupo MTK2. Já o fosfato mostrouse significantemente elevado nos períodos de 7 e 21 dias no grupo MTK2 em relação aos demais grupos. FAL e TRAP total não apresentaram níveis plasmáticos significativamente diferentes comparando-se os grupos e períodos. Considerando os resultados obtidos, concluiu-se que o MTK exerceu efeito tempo-dose-dependente, acelerando o processo de reparo ósseo intramembranoso alveolar e interferindo nos níveis plasmáticos de cálcio e fosfato no presente modelo animal(AU)

Arachidonic acid metabolites are known to play an important role in inflammatory processes and in bone metabolism. However, the role of these products on alveolar bone repair post tooth extraction remains to be explored, especially leukotrienes, derived from 5-lipoxygenase (5-LO). The present study aims to analyze the time-doseresponse effects of the drug montelukast (MTK), a potent type 1 leukotriene cystenyl antagonist (CisLT1Rs), in the alveolar repair process after extraction in male 129Sv/Ev mice. For this purpose, 70 young male mice were used, divided into four groups: C - Control Group (no treatment); VC - Vehicle Control Group, treated with 20 µL of 0.9% SF; MTK2 - treated with 2mg / Kg of MTK and MTK4 - treated with 4mg / Kg of MTK. The animals of the CV, MTK2 and MTK4 groups were treated daily orally (V.O.), starting 24 hours before the surgical procedure, continuing until the end of the experimental periods of 7, 14 and 21 days postoperatively. At the end of the experimental periods, the animals were euthanized for blood collection for serum markers as calcium, phosphate, tartrate-resistant acid phosphate (TRAP) and alkaline phosphatasis (FAL), and to removal of the right maxilla containing the dental socket to be analyzed under computed microtomography (microCT) and histopathology. The results obtained were subjected to statistical tests considering a confidence level of 5%. Results revealed an increase in BV/TV for MTK vs. C and CV groups, in both 14 and 21 days time points. Of note, this increase was higher in MTK4 than in the MTK2 at 14 days. Considering Tb.Th, both MTK2 e MTK4 groups presented positive effects in the BV/TV and Tb.Th increase when compared to controls groups (C and CV) at 21 days. A decrease in BV/TV and Tb.Th was observed in CV compared to C, as a negative effect of animal manipulation. As observed in H&E sections, both MTK2 and MTK4 experimental groups presented an early bone repair in comparison with C group from 7 to 21 days. CV group presented a slight delayed bone healing compared to C. Levels of calcium was increased in MTK4 in comparison to C and MTK2 at 7 and 21 days. Phosphate was significantly elevated at 7 and 21 days in MTK2 in comparison to the other groups. Despite of beneficial effects on observed on morphological levels on sites of healing (microCT and HE), no significant changes were found for bone markers of remodeling in blood plasma (FAL and TRAP). Taken together, these results indicate that MTK induced early bone healing post tooth extraction in 129Sv/Ev mice. Thus, the inhibition of CysLT is suggested to exert a positive influence on intramembranous bone repair post tooth extraction(AU)

In vitro antioxidant, antilipoxygenase and antimicrobial activities of extracts from seven climbing plants belonging to the Bignoniaceae / 中西医结合学报

<p><b>OBJECTIVES</b>This study aimed to evaluate the in vitro antioxidant capacity, to determine the anti-inflammatory effect due to lipoxygenase inhibition and to test the antimicrobial activity of ethanolic extracts from leaves of seven climbing species belonging to the Bignoniaceae family. These species are Adenocalymma marginatum (Cham.) DC., Amphilophium vauthieri DC., Cuspidaria convoluta (Vell.) A. H. Gentry, Dolichandra dentata (K. Schum.) L. G. Lohmann, Fridericia caudigera (S. Moore) L. G. Lohmann, Fridericia chica (Bonpl.) L. G. Lohmann and Tanaecium selloi (Spreng.) L. G. Lohmann.</p><p><b>METHODS</b>The antioxidant activity was evaluated using three methods, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power. Lipoxygenase-inhibiting activity was assayed spectrophotometrically; the result was expressed as percent inhibition. The antimicrobial activity was assessed using the agar disk diffusion method. Minimal inhibitory concentration (MIC) and minimal bactericidal/fungicidal concentration were also determined for each extract against 12 pathogenic bacterial strains of Staphylococcus aureus and seven fungal strains of the Candida genus. The identification of the major compounds present in the most promising extract was established by high-performance liquid chromatography-tandem mass spectrometry.</p><p><b>RESULTS</b>C. convoluta, F. caudigera, and F. chica exhibited the best antioxidant activity by scavenging DPPH and ABTS radicals and reducing Fe ion. These extracts showed a notable inhibition of lipoxygenase. F. caudigera was found to have the lower MIC value against S. aureus strains and six Candida species. The extracts of F. caudigera and C. convoluta were active even against methicillin-resistant S. aureus. C. convoluta had higher total phenol content, better antioxidant activity and superior anti-inflammatory and antimicrobial activity. The main phenolic compounds found in this extract were coumaric and hydroxybenzoic acid derivatives and glycosylated and nonglycosylated flavones.</p><p><b>CONCLUSION</b>Most of the extracts exhibited antioxidant activity as well as in vitro inhibition of lipoxygenase. The excellent antimicrobial activity of T. selloi and F. chica supports their use in traditional medicine as antiseptic agents. The extracts of F. caudigera and C. convoluta, both with notable biological activities in this study, could be used as herbal remedies for skin care. In addition, this study provides, for the first time, information about phenolic compounds present in C. convoluta.</p>

Inhibition of cyclooxygenase (COX) and 5-lipoxygenase (5-LOX) by D-005 (A lipid extract of Acrocomia crispa fruits) / [Inhibición de la ciclooxigenasa (COX) y la 5-lipooxigenasa (5-LOX) por el D-005 (extracto lipidico del fruto de la Acrocomia crispa)

This study was aimed to investigate whether the a lipid extract from Acrocomia crispa fruits (D-005) inhibits COX and 5-LOX enzyme activities in vitro. This study demonstrates that D-005 inhibits markedly and in a dose dependent manner COX-2 and 5-LOX activities. The dual inhibition of COX-2 and 5-LOX supports further research on the potential anti-inflammatory effect of D-005.

El objetivo de este estudio fue investigar si el extracto lipídico de los frutos de Acrocomia crispa (D-005) inhibe in vitro las actividades de las enzimas COX y 5-LOX. Este estudio demuestra que el D-005 inhibe marcadamente y de manera dosis dependiente las actividades de la COX-2 y 5-LOX. La inhibición dual de la COX-2 y 5-LOX soportan futuras investigaciones sobre el potencial efecto anti-inflamatorio del D-005.

Pedalitin from isodon japonica, an inactivation of soybean lipoxygenase-1 / Pedalitina de isodon japonica, un inactivador de la lipo oxigenasa-1 de poroto de soya

Pedalitin, isolated from the aerial part of Rabdosia japonica (Labiatae), inhibited soybean lipoxygenase-1 (EC 1.13.11.12, Type I) with an IC50 of 152.5 uM. The progress curves for an enzyme reaction, pedalitin inactivate the lipoxygenase-1 in a time dependent, irreversible manner, exhibiting kinetics with a kinact/KI of 59.6 +/- 10 mM-1min-1. In the pseudoperoxidase activity, pedalitin is very slowly oxidized by the soybean lipoxygenase-1 catalyzed decomposition of lipid hydroperoxides.

Pedalitina, aislada de las partes aereas de Rabdosia japonica inhibió a la lipooxigenasa-1 (EC 1.13.11.12 tipo I) con un IC50 de 152.5 uM. La curva de progreso para una acción enzimática, pedalitina inactivó a la lipooxigenasa-1 de una manera dependiente del tiempo, de una manera irreversible, exhibiendo una cinética con una kinact/KI de 59.6 +/- mM-1min-1. En la actividad pseudoperoxidasa, pedalitina es oxidada lentamente por la descomposición de la lípido hidroperóxido de la lipooxigenasa-1 de poroto de soya.

The role of leukotriene modifier in pediatric allergic disease

Leukotriene (LT) modifiers are composed of leukotriene receptor antagonists and 5-lipoxygenase inhibitors. LTs, C4, D4, and E4 are collectively termed cysteinyl LTs and best are the characterized receptors for cysLTs are cysLT1 and cysLT2. cysLT1 ligation mediates sustained bronchial contraction, mucosal secretion, and edema, which are central to the pathogenesis of asthma. cysLT2 ligation is thought to contribute to edema, inflammation, and tissue fibrosis in asthma. LT modifiers attenuate bronchoconstriction responses and exert anti-inflammatory effects, reflected by reduced eosinophil counts in the peripheral blood, sputum, and bronchoalveolar lavage fluid of asthmatic patients. Inhaled corticosteroids are generally superior to LT modifiers as a first-line controller. However, LT modifiers are easy to administer, have good compliance, and have excellent safety. LT modifiers are recommended for asthmatic children aged < or =5 years as a first-line controller. The Japanese Guideline for Childhood Asthma recommends LT modifiers, as a first-line controller prior to inhaled corticosteroid for children aged <2 years. LT modifiers can improve asthma control as add-on therapy with ICS. They can also be effective for exercise-induced asthma. LT modifiers are recommended for the treatment of allergic rhinitis in combination with H1-antihistamines or as a first-line drug for patients who cannot or do not wish to use intranasal corticosteroids. LT modifiers can also be considered for add-on therapy in the treatment of chronic urticaria, atopic dermatitis, and other allergic diseases.

Efecto in vitro del policosanol (alcoholes de la cera de Saccharum officinarum L) sobre la enzima 5-lipooxigenasa / In vitro effects of policosanol (Saccharum officinarum L wax alcohols) on the 5-lipooxygenase enzyme

INTRODUCTION: policosanol, a mixture of high molecular weight aliphatic alcohols purified from sugarcane with octacosanol as the main component, shows cholesterol-lowering and antiplatelet effects in addition to an inhibitory effect on type I cicloxygenase. OBJECTIVE: to determine whether policosanol may inhibit 5-LOX enzyme activity in vitro. METHODS: effects on 5-LOX enzyme activities were assessed in rat blood polymorphonuclear leukocytes. Vehicle or Policosanol suspensions (0.6 to 6 000 µg/mL) were added to tubes containing the reaction mix and then absorbance changes at 234 nm were measured. RESULTS: added Policosanol inhibited in vitro 5-LOX activity by 30 percent, which was not a significant figure but depended on the concentration(r= 0.992; p< 0.05); it was 1 250 µg/mL. CONCLUSIONS: policosanol did not significantly inhibit 5-LOX enzyme activity in rat PMNL preparations, so that it does not seem to be a dual inhibitor of COX and-LOX enzymes. This result differs from that found for beeswax alcohols and underlines the different effects of the mixtures of long-chain fatty alcohols purified from the sugarcane and the beeswax(AU).

IINTRODUCCIÓN: el policosanol es una mezcla de alcoholes alifáticos aislados y purificados de la caña de azúcar cuyo componente mayoritario es el octacosanol, con efecto sobre la reducción de colesterol y antiagregante plaquetario, además inhibe la ciclooxigenasa (COX) tipo 1. OBJETIVO: determinar el poder de inhibición del policosanol en la actividad de la enzima 5-LOX in vitro. MÉTODOS: el efecto sobre la actividad de la enzima 5-LOX se determinó en leucocitos polimorfonucleares obtenidos de sangre de ratas. Se añadieron vehículo o suspensiones de policosanol (0,6 a 6 000 µg/mL) a tubos que contenían la mezcla de reacción y se medió el cambio de absorbancia a 234 nm. RESULTADOS: la adición de policosanol inhibió in vitro la actividad de la 5-LOX en un 30 por ciento que no fue significativo pero sí dependiente de la concentración (r= 0,992; p< 0,05), inhibición esta que alcanzó 1 250 µg/mL. CONCLUSIÓN: el policosanol no inhibió significativamente la actividad de la enzima 5-LOX en preparación de polimorfonucleares de ratas, por lo que no es un inhibidor dual de las enzimas. Este resultado difiere del encontrado para los alcoholes de la cera de abeja y subraya la diferencia de los efectos hallados entre las mezclas de alcoholes alifáticos de cadenas largas purificados de la caña de azúcar y la cera de abeja(AU)

Pharmacological basis of the use of Acortus calamus L. in inflammatory diseases and underlying signal transduction pathways / Bases farmacológicas del uso de Acorus calamus L. en enfermedades inflamatorias y las vías de transducción de señales subyacentes

Acorus calamus L. is used as anti-inflammatory remedy in traditional system of medicine in Pakistan and India. This study was designed to explore the anti-inflammatory mechanism of Acorus calamus L. and its underlying signaling pathways. Aqueous, butanolic and n-hexane fractions of Acorus calamus were tested against cyclooxygenase (COX) and lipoxygenase (LOX) mediated eicosanoids production by arachidonic acid (AA). Butanolic fraction of Acorus calamus, but not the aqueous and n-hexane fractions, inhibited the COX mediated production of thromboxane B2 (TXB2) and liopxygenase product 1 (LP1) -a metabolite of LOX pathway. 12-(hydroxyeicosatetraenoic acid) HETE- another product of the LOX pathway was unaffected by all three fractions. Butanolic fraction of Acorus calamus showed strong inhibition against AA-induced platelet aggregation. Investigation of the underlying signaling pathways revealed that butanolic fraction inhibited phospholipase C (PLC) pathway in platelets, most probably acting on protein kinase C (PKC). Aqueous and n-hexane fractions of Acorus calamus were not effective against any platelet agonist. This study shows that butanolic fraction of Acorus calamus possesses components that inhibit AA metabolism and platelet aggregation through multiple pathways.

Acorus calamus L. se utiliza como remedio anti-inflamatorio en el sistema tradicional de la medicina en Pakistán y la India. Este estudio fue diseñado para explorar el mecanismo anti-inflamatorio de Acorus calamus L. y sus vías de señalización subyacentes. Fracciones acuosa, butanólica y de n-hexano de Acorus calamus se ensayaron frente a la ciclooxigenasa (COX) y de la lipoxigenasa (LOX) mediada por la producción de eicosanoides por el ácido araquidónico (AA). La fracción butanólica de Acorus calamus, pero no las fracciones acuosas y de n-hexano, inhibieron la producción de COX mediada por tromboxano B2 (TXB2) y el producto lipoxigenasa 1 (LP1) - un metabolito de la vía de LOX, 12 - (ácido hidroxieicosatetraenoico) HETE - otro producto de la ruta de LOX no fue afectado por las tres fracciones. La fracción butanólica de Acorus calamus mostró una fuerte inhibición contra la agregación plaquetaria inducida por AA. La investigación de las vías de señalización subyacentes reveló que la fracción butanólica inhibió la fosfolipasa C (PLC) y la vía en las plaquetas, probablemente actuando sobre la proteína quinasa C (PKC). Fracciones acuosas y de n - hexano de Acorus calamus no fueron eficaces contra ningún agonista de plaquetas. Este estudio muestra que la fracción butanólica de Acorus calamus posee componentes que inhiben el metabolismo del AA y la agregación plaquetaria a través de múltiples vías.

Zileuton, a 5-lipoxygenase inhibitor, attenuates mouse microglial cell-mediated rotenone toxicity in PC12 cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To examine the effect of a selective inhibitor of 5-lipoxygenase (5-LOX) zileuton on microglia-mediated rotenone neurotoxicity.</p><p><b>METHODS</b>The supernatant from different concentrations of rotenone-stimulated mouse microglia BV2 cells was used as the conditioned media (CM) for PC12 cells. The viability of PC12 cells was determined by MTT assay and lactate dehydrogenase (LDH) release. Cell death was observed by LDH release and double fluorescence staining with Hoechst/propidiumiodide (PI). The effect of zileuton on microglia-mediated rotenone toxicity was evaluated by the above methods.</p><p><b>RESULTS</b>Rotenone at 1-10 nmol/L was nontoxic to PC12 cells directly. However, the CM from BV2 cells that were treated with rotenone (1-10 nmol/L) resulted in toxicity of PC12 cells. The BV2 CM which stimulated with rotenone (1-10 nmol/L) induced morphological changes, reduced cell viability, and increased LDH release and cell necrosis in PC12 cells. Pretreatment of BV2 cells with the 5-LOX inhibitor zileuton (0.01-1 μmol/L) protected PC12 cells from the microglia-mediated rotenone toxicity.</p><p><b>CONCLUSION</b>The 5-LOX inhibitor zileuton effectively attenuates microglia-mediated rotenone toxicity in PC12 cells. These results suggest that 5-LOX pathway may be involved in neuronal death induced by microglial inflammation.</p>

Garden rue inhibits the arachidonic acid pathway, scavenges free radicals, and elevates FRAP: role in inflammation / 中国天然药物

AIM@#In the present study, the anti-inflammatory and antioxidant activities of the methanol extract of Ruta graveolens leaves (RG-M) were evaluated using various in vivo and in vitro models.@*METHOD@#For anti-inflammatory activity, RG-M was administered by the oral route (p.o.) in a carrageenan-induced paw edema model, and by the intraperitoneal route (i.p.) in an exudative inflammation model. In vitro inhibition of cyclooxygenase and lipoxygenase enzymes was evaluated. In vitro antioxidant activity was also examined. Endogenous antioxidant status was further evaluated by ferric reducing ability of plasma model.@*RESULTS@#RG-M showed maximum inhibition of carrageenan-induced edema (100 mg·kg⁻¹ - 33.36%; 200 mg·kg⁻¹ - 45.32% and 400 mg·kg⁻¹ - 56.28%). In the exudative inflammation model, a significant reduction in leukocyte migration (200 mg·kg⁻¹ - 54.75% and 400 mg·kg⁻¹ - 77.97%) and protein exudation (200 mg·kg⁻¹ - 31.14% and 400 mg·kg⁻¹ - 49.91%) were observed. RG-M also exhibited inhibition of COX-1 (IC50 182.27 μg·mL⁻¹) and COX-2 (IC50 190.16 μg·mL⁻¹) as well as 5-LOX (IC50 215.71 μg·mL⁻¹). Antioxidant activity was significant with improved endogenous antioxidant status.@*CONCLUSION@#The results demonstrated the anti-inflammatory and antioxidant activity of RG-M with potent inhibitory effects on the arachidonic acid pathways.

Expression of 5-lipoxygenase in hippocampal CA1 neuronal damage following global cerebral ischemia in rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine 5-lipoxygenase (5-LOX) expression and the effect of zileuton, a selective 5-LOX inhibitor,on hippocampal neuron injury induced by global cerebral ischemia in rats.</p><p><b>METHODS</b>Global cerebral ischemia was induced by bilateral common carotid artery occlusion combined with hypotension in rats. 5-LOX expression was detected by Western blot analyses and 5-LOX localization was visualized by immunohistochemistry and double immunofluorescence methods. The 5-LOX inhibitor zileuton (10, 30, 50 mg/kg) was orally administered for 3 d after ischemia.</p><p><b>RESULTS</b>The 5-LOX expression was increased in the ischemic hippocampus on d1-7 (peaked at d3), and 5-LOX protein was primarily localized in neurons and translocated to the nuclei in the hippocampal CA1 region after ischemia. The 5-LOX inhibitor zileuton (30, 50 mg/kg) reduced ischemia-induced hippocampal neurons death 3d after ischemia.</p><p><b>CONCLUSION</b>5-LOX is involved in global cerebral ischemic damage in rats, and the 5-LOX inhibitor zileuton has a protective effect on neuronal damage in the rat hippocampus following global cerebral ischemia.</p>

Cyclooxygenases, lipoxygenases, their targeted drugs and the prevention of Alzheimer's disease / 药学学报

Many studies have shown that chronic inflammation occurs in the brain of patients with Alzheimer's disease (AD). It is well known that long-term administration of non-steroidal anti-inflammatory drugs (NSAIDs) can alleviate the cognitive decline of AD patient and elderly. Several inflammatory cytokines produced in the metabolism of arachidonic acid (AA) are closely related to inflammatory diseases. Lipoxygenases (LOXs) and cyclooxygenases (COXs) play a crucial role in the AA network, the products eicosanoids have an important impact on the progression of AD. Although there are many arguments and conflicting evidence, currently LOXs and COXs are still the hot topics in the research on AD pathogenesis and drug development. Here, we review the progress in research on COXs and LOXs, including their actions on CNS and their association with AD, and explore the feasibility of LOXs and COXs as targets for the drugs to prevent and/or treat AD.

Synthesis, characterization and biological screening of N-substituted derivatives of 5-benzyl-1,3,4-oxadiazole-2yl-2''-sulfanyl acetamide

A series of new N-substituted derivatives of 5-benzyl-1, 3, 4-oxadiazole-2yl-2''-sulfanyl acetamide [6a-n] were synthesized in three phases. The first phase involved the sequentially converting phenyl acetic acid into ester, hydrazide and finally cyclized in the presence of CS[2] to afford 5-benzyl-1, 3, 4-oxadiazole-2-thiol. In the second phase N-substituted-2-bromoacetamides were prepared by reacting substituted amines with bromoacetyl bromide in basic media. In the third phase, 5-benzyl-1,3,4-oxadiazole-2-thiol was stirred with N-substituted-2-bromoacetamides in the presence of N,N-dimethyl formamide [DMF] and sodium hydride [NaH] to get the target compounds. Spectral techniques were used to confirm the structures of synthesized compounds. Synthesized compounds were screened against butyrylcho linesterase [BChE], acetylcholinesterase [AChE], and lipoxygenase enzymes [LOX] and were found to be relatively more active against acetylcholinesterase

Synthesis of biologically active O-substituted derivatives of 1-[[3, 5-dichloro-2-hydroxyphenyl]sulfonyl]piperidine

In the present work, a series of 2-O-substituted derivatives of 1-[[3,5-dichloro-2-hydroxy phenyl] sulfonyl]piperidine [5a-j] were synthesized. These derivatives were geared up by the coupling of 3,5-dichloro-2-hydroxy benzenesulfonyl chloride [1] with piperidine [2] under dynamic pH control in aqueous media to form parent compound 1-[[3,5-dichloro-2-hydroxyphenyl]sulfonyl]piperidine [3], followed by the substitution at oxygen atom with different electrophiles [4a-j] in the presence of sodium hydride [NaH] and dimethyl formamide [DMF] to give a series of Osubstituted derivatives of sulfonamides bearing piperidine nucleus 5a-j. The synthesized O-substituted sulfonamides were spectrally characterized. The bioactivity of all the synthesized compounds were evaluated against lipoxygenase [LOX], acetylcholinesterae [AChE] and butyrylcholinesterase [BChE] enzymes and found to be having talented activity against butyrylcholinesterase enzyme

Benzoxazole Derivative B-98 Ameliorates Dextran Sulfate Sodium-induced Acute Murine Colitis and the Change of T Cell Profiles in Acute Murine Colitis Model / 대한소화기학회지

BACKGROUND/AIMS: The unique role of enzyme 5-lipoxygenase (5-LO) in the production of leukotrienes makes it a therapeutic target for inflammatory bowel disease (IBD). The aim of this study was to evaluate the effects of B-98, a newly synthesized benzoxazole derivatives and a novel 5-LO inhibitor, in a mouse model of IBD induced by dextran sulfate sodium (DSS). METHODS: C57BL/6 mice were randomly assigned to four groups: normal control, DSS colitis (DSS+saline), low dose B-98 (DSS+B-98 20 mg/kg) and high dose B-98 (DSS+B-98 100 mg/kg). B-98 was administered with 3% DSS intraperitoneally. The severity of the colitis was assessed via the disease activity index (DAI), colon length, and histopathologic grading. The production of inflammatory cytokines interleukin (IL)-6 was determined by RT-PCR. Th cells were examined for the proportion of Th1 cell, Th2 cell, Th9 cell, Th17 cell and Treg cell using intracellular cytometry. RESULTS: The B-98 group showed lower DAI, less shortening of the colon length and lower histopathologic grading compared with the DSS colitis group (p<0.01). The expression of IL-6 in colonic tissue was significantly lower in the B-98 groups than the DSS colitis group (p<0.05). The cellular profiles revealed that the Th1, Th9 and Th17 cells were increased in the DSS colitis group compared to the B-98 group (p<0.05). CONCLUSIONS: Our results suggest that acute intestinal inflammation is reduced in the group treated with B-98 by Th1, Th9 and Th17 involved cellular immunity.

Carbohydrate content and antioxidative potential of the seed of three edible indica rice (Oryza sativa L.) cultivars.

Rice (Oryza sativa L.) grains or seeds are known to lose much of their nutrient and antioxidant contents, following polishing. The current study was undertaken to evaluate and compare the carbohydrate content and antioxidant parameters in the unpolished and polished seeds of three edible indica rice cultivars, namely Swarna (SW), the most popular indica rice cultivar in India and aromatic or scented cultivars Gobindobhog (GB) and Pusa Basmati (PB). While both the sucrose and starch content was the maximum in PB seeds (both unpolished and polished), the amylose content was the highest in SW polished seeds. SW polished seeds were superior as compared to GB and PB cultivars in terms of total antioxidant capacity, DPPH radical scavenging and Fe(II) chelation potential, as well as the highest lipoxygenase (LOX) inhibition or H2O2 scavenging potential, probably due to the maximum accumulation of total phenolics and flavonoids, the two important antioxidants. The reducing power ability was, however, identical in both SW and GB polished seeds. The PB polished seeds were more potent in superoxide and hydroxyl scavenging, whereas GB in nitric oxide (NO) scavenging. The common observation noted after polishing of seeds was the reduction in the level of carbohydrates and antioxidant potential, though the extent of reduction varied in the three cultivars. The only exception was GB, where there was no alteration in NO scavenging potential even after polishing. Our study showed the better performance of SW polished seeds with respect to higher amylose content and majority of the tested parameters governing antioxidant capacity and radical scavenging potential, thus highlighting the greater dietary significance of SW over the other two cultivars.

In vitro anti-inflammatory effects of naturally-occurring compounds from two Lauraceae plants

The in vitro anti-inflammatory effects of seven known lignans and one dihydrochalcone isolated from the leaves of two Lauraceae species (Pleurothyrium cinereum and Ocotea macrophylla), were evaluated through the inhibition of COX-1, COX-2, 5-LOX and the aggregation of rabbit platelets induced by PAF, AA and ADP. (+)-de-4"-O--methylmagnolin 4 was found to be a potent COX-2/5-LOX dual inhibitor and PAF-antagonist (COX-2 IC50 2.27 µM; 5-LOX IC50 5.05 µM; PAF IC50 2.51 µM). However, all compounds exhibited an activity at different levels, indicating good anti-inflammatory properties to be considered in further structural optimization studies.

Os efeitos anti-inflamatórios in vitro de sete conhecidos lignanos e uma dihidrocalcona isolados das folhas de duas espécies da família Lauraceae (Pleurothyrium cinereum e Ocotea macrophylla) foram avaliados por meio da inibição da COX1, COX-2, 5-LOX e agregação de plaquetas de coelhos induzida por PAF, AA e ADP. A (+)-4"-O-metilmagnolina-4 foi encontrada como mais potente inibidora tanto da COX-2 quanto de 5-LOX e antagonista de PAF (COX-2 IC50 2,27 µM; 5- LOX IC50 5,05 µM; PAF IC50 2,51 µM). Entretanto, todos compostos mostram uma atividade em intensidades diferentes, indicando boas propriedades anti-inflamátorias a serem consideradas para futuros estudos de modificações e otimização estruturais.

High-throughput screening assays for cyclooxygenase-2 and 5-lipoxygenase, the targets for inflammatory disorders.

High-throughput screening (HTS) involves testing of compound libraries against validated drug targets using quantitative bioassays to identify ‘hit’ molecules that modulate the activity of target, which forms the starting point of a drug discovery effort. Eicosanoids formed via cyclooxygenase (COX) and lipoxygenase (LOX) pathways are major players in various inflammatory disorders. As the conventional non-steroidal anti-inflammatory drugs (NSAIDs) that inhibit both the constitutive (COX-1) and the inducible (COX-2) isoforms have gastric and renal side effects and the recently developed COX-2 selective anti-inflammatory drugs (COXIBs) have cardiac side effects, efforts are being made to develop more potent and safer anti-inflammatory drugs. Current assay methods for these enzymes, such as oxygraphic, radioisotopic, spectrophotometric etc. are not compatible for screening of large number of compounds as in drug discovery programs. In the present study, HTS-compatible assays for COX-1, COX-2 and 5-LOX were developed for screening of compound libraries with the view to identify potential anti-inflammatory drug candidates. A spectrophotometric assay involving co-oxidation of tetramethyl-p-phenylene diamine (TMPD) during the reduction of prostaglandin G2 (PGG2) to PGH2 was adopted and standardized for screening of compounds against COX-1 and COX-2. Similarly, the HTS-compatible FOX (ferrous oxidation-xylenol orange) based spectrophotometric assay involving the formation of Fe3+/xylenol orange complex showing absorption in the visible range was developed for screening of compounds against 5-LOX.

5-lipoxygenase is involved in rotenone-induced injury in PC12 cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine whether 5-lipoxygenase (5-LOX) is involved in rotenone-induced injury in PC12 cells, which is a cell model of Parkinson disease.</p><p><b>METHODS</b>After rotenone treatment for various durations, cell viability was determined by colorimetric MTT reduction assay, and 5-LOX translocation was detected by immunocytochemistry. The effect of 5-LOX inhibitor zileuton was also investigated.</p><p><b>RESULT</b>Rotenone (0.3-30 μmol/L) induced PC12 cell injury, and zileuton (3-100 μmol/L) attenuated this injury. Rotenone also time-and concentration-dependently induced 5-LOX translocation into the nuclear envelope, and zileuton (1-30 μmo/L) significantly inhibited rotenone-induced 5-LOX translocation.</p><p><b>CONCLUSION</b>5-LOX is involved in rotenone-induced injury in PC12 cells, and 5-LOX inhibitor zileuton can reduce rotenone-induced 5-LOX activation and cell injury.</p>