Screening of phytocompounds, molecular docking studies, and in vivo anti-inflammatory activity of heartwood aqueous extract of Pterocarpus santalinus L.f

Objective:To evaluate the anti-inflammatory potential of aqueous extract of Pterocarpus santalinus L.f. heartwood using molecular docking and in vivo experiment. Methods:An aqueous extract of Pterocarpus santalinus heartwood was prepared using a Soxhlet apparatus. Phytocompounds in the extract were tentatively identified using high-resolution mass spectrometry. Molecular docking experiments were carried out to evaluate the binding affinity of selected compounds, phloridzin to cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), prostaglandin E synthase-1 (PGES-1) and 5-lipoxygenase (5-LOX). Anti-inflammatory potential was evaluated by carageenan induced paw edema model in rats. Results:The presence of major component phloridzin along with quercetin, parthenin, ginkgolide B, picrotoxinin, usnic acid, octopine, and epigallocatechin was detected in the extract. Molecular docking study showed that phloridzin inhibited COX-1, COX-2, PGES-1 and 5-LOX with more affinity than ibuprofen and paracetamol. Pterocarpus santalinus heartwood extract at 200 and 400 mg/kg BW showed significant reduction in carageenan-induced hind paw edema in a dose-dependent manner, but the effect was slow when compared with the standard ibuprofen (30 mg/kg p.o.). Conclusions:The study indicated that after clinical trials, the aqueous extract of Pterocarpus santalinus heartwood can be effectively used in phytotherapy to treat inflammation.

Effects of baicalin-gardenoside pairing on 5-lox/CysLTs/CysLT pathway of cerebral ischemic injury rats / 中成药

AIM To study the effects of baicalin and gardenoside pairing on 5-lox/CysLTs/CysLT pathway of rats with cerebral ischemia injury.METHODS SD rats randomly assigned into control group,model group,baicalin and gardenoside pairing groups (7 ∶ 3,dosed at 30,45,60 mg/kg,respectively) in their cerebral ischemia recovery period were simulated with permanent middle cerebral artery occlusion (pMCAO) if necessary.After a week of natural recovery,the drugs' pharmacodynamic effects were evaluated by the neurofunctional scoring and HE staining,their impact on the content of CysLTs was determined by ELISA,and the influence on the expression of 5-lox,CysLT1 and CysLT2 was detected by western blot.RESULTS One-week consecutive administration of baicalin and gardenoside pairing contributed to a reduction in infiltration and tissue edema of ischemic cells.If compared with pMCAO group,the baicalin and gardenoside pairing groups were observed with significantly lowered scores of neurologic functioning,inhibited microglia activation,decreased content of CysLTs and down-regulated expression of 5-LOX,CysLT1 and CysLT2 (P ＜ 0.01).CONCLUSION Baicalin and gardenia pairing's alleviating effect to the damage of cerebral ischemia may be associated with its inhibition on microglia activation and 5-lox/CysLTs/CysLT signaling pathway,and thus an inflammatory impairment reduction.

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.

5-LOX inhibitor MK886 suppresses the stemness of human colon cancer HT-29 cells / 医学研究生学报

Objective It remains a controversy whether 5-lipoxygenase (5-LOX) is associated with colon cancer stem cells.This study was to investigate the effect of the 5-LOX inhibitor MK886 in maintaining the stemness of the human colon cancer cell line HT-29.Methods Using CCK-8 assay, we examined the inhibitory effects of different concentrations of MK886 (12.5, 25, 50, 75, 100, and 200 μmol/L) on the colon cancer HT-29 cells cultured in vitro and calculated its half-inhibitory concentration (IC50).Then, we detected the effects of MK886 IC50 on the clone-and sphere-forming abilities of the cells, determined the mRNA expressions of the stemness markers CD133, Lgr5, Oct4 and Ascl2 by real-time PCR after 24 and 48 hours of MK886 IC50 intervention, and measured their protein expressions by Western blotting after 24, 48 and 72 hours of MK886 IC50 intervention.Results The inhibition rates of MK886 on the HT-29 cells at 24 and 48 hours were significantly increased in a time-and dose-dependent manner ([14.99±3.06] and [19.98±0.57]％ at 12.5 μmol/L, [20.46±1.14] and [34.97±6.02]％ at 25 μmol/L, [50.76±5.94] and [66.90±5.74]％ at 50 μmol/L, [66.84±1.77] and [73.11±2.48]％ at 75 μmol/L, [72.67±2.36] and [77.78±3.30]％ at 100 μmol/L, [83.67±0.24] and [84.69±2.24] ％ at 200 μmol/L) as compared with the blank control (0％ and 0％) (P<0.05).The clone-forming rate and number of spheres formed were remarkably lower in the MK886 intervention than in the control group ([10.60±1.71] vs [44.67±3.21]％, P<0.05;6.00±1.60 vs 19.07±2.89, P<0.05).After 24 and 48 hours of MK886 intervention, the mRNA expression of CD133 in the HT-29 cells was markedly up-regulated in comparison with that at 0 hour (0.72±0.10 and 0.39±0.07 vs 1.66±0.33, P<0.05), and so were those of Lgr5, Oct4 and Ascl2 (P<0.05).Conclusion The 5-LOX inhibitor MK886 can inhibit the proliferation and clone-and sphere-forming abilities of human colon cancer HT-29 cells by down-regulating the expressions of the stemness markers and thus suppressing the stemness of the colon cancer stem cells.

Structural optimization and biological evaluation of 1,5-disubstituted pyrazole-3-carboxamines as potent inhibitors of human 5-lipoxygenase

Human 5-lipoxygenase (5-LOX) is a well-validated drug target and its inhibitors are potential drugs for treating leukotriene-related disorders. Our previous work on structural optimization of the hit compound 2 from our in-house collection identified two lead compounds, 3a and 3b, exhibiting a potent inhibitory profile against 5-LOX with IC50 values less than 1 µmol/L in cell-based assays. Here, we further optimized these compounds to prepare a class of novel pyrazole derivatives by opening the fused-ring system. Several new compounds exhibited more potent inhibitory activity than the lead compounds against 5-LOX. In particular, compound 4e not only suppressed lipopolysaccharide-induced inflammation in brain inflammatory cells and protected neurons from oxidative toxicity, but also significantly decreased infarct damage in a mouse model of cerebral ischemia. Molecular docking analysis further confirmed the consistency of our theoretical results and experimental data. In conclusion, the excellent in vitro and in vivo inhibitory activities of these compounds against 5-LOX suggested that these novel chemical structures have a promising therapeutic potential to treat leukotriene-related disorders.

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)

Efecto in vitro del D-003 sobre la actividad enzimática de la 5-lipoxigenasa (5-LOX) / Effect in vitro of D-003 on 5-lipooxygenase (5-LOX) enzyme activity

Introducción: el D-003, mezcla de ácidos alifáticos primarios superiores purificada de la cera de la caña, inhibe la síntesis de colesterol. Trabajos recientes han demostrado que el D-003 resulta efectivo en modelos experimentales de osteoartritis y que inhibe la actividad de las enzimas COX1 y COX2, preferentemente la COX1, sin producir gastrotoxicidad. Ha sido referido que los inhibidores duales de las enzimas COX y 5-LOX presentan efectos antinflamatorios desprovistos de gastrotoxicidad o que incluso, pueden resultar gastroprotectores. De acuerdo con estos antecedentes, el D-003 podría ser un inhibidor dual de dichas enzimas. Objetivo: investigar el efecto in vitro del D-003 sobre la actividad enzimática de la 5-LOX, utilizando la fracción citosólica de leucocitos polimorfonucleares de ratas. Métodos: se utilizaron las condiciones de ensayo siguientes: fracción citosólica (50 µg de proteína) disuelta en solución reguladora borato 0,2 mol/L (pH 9) y ácido linoleico (7,8-250 mmol/L) como sustrato, ensayándose muestras paralelas incubadas con Tween-20/H2O (2 por ciento) (vehículo), D-003 (0,6-6 000 µg/mL) o extracto de Perna canaliculus (50 µg/mL) (sustancia de referencia). Se evaluó la actividad enzimática mediante el cambio de absorbancia a 234 nm producido por la formación de dienos conjugados y medido en espectrofotómetro UV-visible. Resultados: la adición de D-003 produjo una inhibición dosis dependiente de la actividad enzimática de la 5-LOX (r= 0,975; p< 0,05) (CI50= 23,06 µg/mL) in vitro. La magnitud de esta inhibición fue moderada, ya que la inhibición máxima, alcanzada a partir de 1 250 µg/mL, resultó de solo un 30 por ciento. Conclusiones: el estudio demuestra que el D-003 es capaz de inhibir la actividad enzimática de la 5-LOX in vitro, pero moderadamente(AU)

Introduction: D-003, a mix of higher primary aliphatic acids purified from sugarcane, inhibits cholesterol synthesis. Recent studies have demonstrated that D-003 is effective in experimental models of osteoarthritis and inhibits the enzymatic activities of COX1 and COX2, mainly that of COX1, without causing gastrotoxicity. It has been mentioned that the dual inhibitors of COX and 5-LOX enzymes present with anti-inflammatory effects and no gastrotoxicity or even they can have gastroprotective actions. According to this background, D-003 could be a dual inhibitor of COX and 5-LOX enzymes. Objective: to study the effects of D-003 on the enzymatic activity of 5-LOX in vitro, by using the cytosolic fraction of polymorphonuclear leukocytes of rats. Methods: the following testing conditions were used: cytosolic fraction (50 µg of protein) dissolved into 0,2 mol/L borate buffer solution (pH 9) and linoleic acid (7,8-250 mmol/L) as substrate. Parallel samples were incubated with Tween-20/H2O (2 percent) only (vehicle), D-003 (0,6-6 000 µg/mL) or green-lipped mussel (Perna canaliculus) extract (50 µg/mL) (reference substance). The enzymatic activity, evaluated by the conjugated diene formation, was assessed by the absorbance changes at 234 nm (5-LOX) measured in a UV-visible spectrophotometer. Results: by adding D-003, produced a dose dependent (r= 0,975; p< 0,05) (IC50= 23,06 µg/mL) in vitro inhibition of 5-LOX enzyme activity. The size of the inhibition was moderate, since the maximal inhibition, achieved from 1 250 µg/mL) on was only 30 percent. Conclusions: this study demonstrates that D-003 may inhibit in vitro the 5-LOX enzymatic activity, but in a moderate way(AU)

In-vitro and in-vivo anti-inflammatory activity of Andrographis serpyllifolia (Rottl. Ex Vahl.) Wt.

The study was aimed to evaluate the analgesic and anti-inflammatory activity (by both in-vitro and in-vivo) of both chloroform and methanol root extracts of Andrographis serpyllifolia (Rottl. Ex Vahl.) Wt. Methods used for the studies were In-vitro 5-Lipoxygenase inhibition assay and In-vivo measurement of rat paw edema and ear edema in rats, acetic acid induced writhing response and hot plate method in albino mice. Chloroform and methanolic extracts of A. serpyllifolia root have shown moderate potency in inhibiting 5-LOX and shown significant anti-inflammatory activity. Despite the IC50 values are little higher, anti-inflammatory efficacy of these extracts possibly due to other mechanisms apart of 5-LOX inhibition. However, In-vivo anti-inflammatory studies revealed that A. serpyllifolia methanolic extract has shown higher degree of efficacy when compared to the chloroform extract. In terms of analgesic activity in writhing test, methanolic extract has shown more efficacy than chloroform extract. Hence, it is important to isolate the active principles for further testing the anti-inflammatory efficacy.

In-vitro and in-vivo anti-inflammatory activity of Andrographis serpyllifolia (Rottl. Ex Vahl.) Wt.

The study was aimed to evaluate the analgesic and anti-inflammatory activity (by both in-vitro and in-vivo) of both chloroform and methanol root extracts of Andrographis serpyllifolia (Rottl. Ex Vahl.) Wt. Methods used for the studies were In-vitro 5-Lipoxygenase inhibition assay and In-vivo measurement of rat paw edema and ear edema in rats, acetic acid induced writhing response and hot plate method in albino mice. Chloroform and methanolic extracts of A. serpyllifolia root have shown moderate potency in inhibiting 5-LOX and shown significant anti-inflammatory activity. Despite the IC50 values are little higher, anti-inflammatory efficacy of these extracts possibly due to other mechanisms apart of 5-LOX inhibition. However, In-vivo anti-inflammatory studies revealed that A. serpyllifolia methanolic extract has shown higher degree of efficacy when compared to the chloroform extract. In terms of analgesic activity in writhing test, methanolic extract has shown more efficacy than chloroform extract. Hence, it is important to isolate the active principles for further testing the anti-inflammatory efficacy.

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.

Synthesis and anti-inflammatory activity of p-(sulfamyl) benzylidene-linked heterocyclic ketone derivatives / 中国药科大学学报

Aim: To study the synthesis and anti-inflammatory activity of p-(sulfamyl) benzylidene-linked hetero-cyclic ketone derivatives. Methods: A series of p-(sulfamyl) benzylidene-linked heterocyclic ketone derivatives were synthesized. Anti-inflammatory activity was evaluated against xylene-induced ear oedema in mice and against carrageenan-induced paw oedema in rats. Gastrointestinal side effects in the rats were also examined after continu-ous introgastric administration of these compounds once daily for 7 days. Results: Twelve compounds( LHZ-101-LHZ-112) were synthesized and their structures were confirmed by IR, ~1H NMR, MS and elemental analysis. LHZ-105, LHZ-106 and LHZ-111 exhibited marked anti-inflammatory activity in xylene-induced mice ear swelling model. LHZ-106 and LHZ-111 showed significant anti-inflammatory activity in carrageenan-induced rat paw ede-ma model. LHZ-105, LHZ-106 and LHZ-111 had less gastrointestinal side effects than diclofenac sodium and CI-1004. Conclusion: These results suggest that some of these compounds have the potential for anti-inflammatory activity with few gastrointestinal side effects.

Inhibition of gastric cancer cells invasion and metastasis by Darbufelone in vitro / 重庆医科大学学报

Objective:To study the effects of Darbufelone on the invasion and metastasis of human gastric cancer cells SGC-7901 in vitro. Methods:The adhesion ability was determined by MTT.Invasion and metastasis was observed in Transwell Chamber membrane invasion culture system.The capability of SGC-7901 cell condition medium in inducing endothelial cell to form blood vessel in 24 well plate was detected.RT-PCR and cel1-immunohistochemistry were used to observe the expressions of OPN and MMP-2.Results:In the membrane invasion culture system,the numbers of invading and migrating SGC-7901 cells were significantly lower in Darbufelone treated groups compared with those in the control group(sP

Zileuton,a 5-LOX inhibitor,inhibits proliferation and induces apoptos is of SGC-7901 gastric cancer cells / 重庆医科大学学报

Objective:To evaluate the efficacy of 5-LOX inhibitor zileuton on the proliferation and apoptosis of human gastric carcinoma cell line SGC-7901,and to explore the mechanisms of apoptosis induced by zileuton.Methods:The proliferation of SGC-7901 cells was detected by thiazolyl blue tetrazolium bromide(MTT)assays.TUNEL staining was used to evaluate the apoptosis of SGC-7901 cells.The changes of Bax,Bcl-2 and caspase-3 mRNA expression were studied by reverse transcription polymerase chain reaction.Bax,Bcl-2 and caspase-3 protein expression were analyzed by immunocytochemistry in SGC-7901 cells treated with zileuton.Results:Zileuton can decrease the proliferation significantly in a dose-dependent and time-dependent manner in SGC-7901 cells.TUNEL staining showed that the apoptosis rate was elevated with the increase of zileuton concentration(P

Expression of 5-LOXmRNA and VEGFmRNA in pancreatic cancer and its clinical significance / 中国普通外科杂志

Objective To investigate the expression of 5-LOX in pancreatic cancer tissue and the relationship between 5-LOX expression and expression of VEGF.Methods The expression of 5-LOXmRNA,(VEGFmRNA) in 35 pancreatic cancer fresh tissue samples were detected by semi-quantitive reverse(transcriptase)-polymerase chain reaction method.Results Expression of 5-LOXmRNA,VEGFmRNA in(pancreatic) cancer tissue were 74.3%,60% respectively,and the expression was correlated to the with(clinical) stages of the tumor;also expression of VEGFmRNA was correlated to the differentiation of the tumor.Expression of 5-LOXmRNA and VEGFmRNA were synergetic in pancreatic cancer(P

Expression of 5-LOX gene and its inhibitory effect in human pancreatic cancer cell / 中国现代普通外科进展

Objective:To detect the expression of 5-LOX and observe its inhibitory effect in humane pancreatic cancer cell.Methods:5-LOX mRNA and protein expression were analyzed by RT-PCR and immunocytochemical staining as well as images analyzing system,and the effect of MK886 was tested.Results:MK886 effectively down-regulated the expression of 5-LOX mRNA.Immunocytochemical staining and images analysis confirmed that the expression 5-LOX protein in cell treated with MK886 was lower than untreated control cell.Conclusion:Inhibitor of 5-LOX can effectively prevent the target mRNA into protein product,thus block or weaken the expression of 5-LOX.

The expressions of 5-LOX mRNA and COX-2 mRNA in pancreatic cancer and their clinical significance / 中国普通外科杂志

0.05).There was correlation between the expression of 5-LOX and TNM stages.The expression of 5-LOX in stages III~IV of pancreatic cancer was markedly higher than that in stages I~II of pancreatic cancer(P

Effect of darbufelone on the proliferation and apoptosis of SGC-7901 cell line / 中国药理学通报

Aim To investigate the effects of dual COX-2/5-LOX inhibitor darbufelone on the inhibition of gastric adenocarcinoma in vitro.Methods MTT assay was used to detect the inhibition of gastric adenocarcinoma cell line SGC-7901 by darbufelone in different concentrations and at different times.TUNEL staining was used to evaluate the apoptosis of SGC-7901 cells.The changes of 5-LOX and COX-2 mRNA expression were studied by reverse transcription polymerase chain reaction.5-LOX and COX-2 protein expressions were analyzed by immunocytochemistry in SGC-7901 cells.Results darbufelone could decrease the proliferation significantly in a dose-dependent and time-dependent manner in SGC-7901 cells.After treating SGC-7901 with darbufelone at concentration of 1.5?10-5,1.0?10-5,5?10-6 mol?L-1 for 72 hs,apoptosis index(%) of SGC-7901 was(30.3?2.1)%,(23.0?2.0)%,(15.0?1.5)%,respectively,which was significantly higher than(0.6?0.1)% that in control group(P