Sulforaphane regulates macrophage glycolysis and inhibits the progression of diabetic nephropathy by modulating the ALOX5/NF-κB signaling pathway / 安徽医科大学学报

Objective @# To investigate the effects of sulforaphane (SFN) in regulating the macrophage glycolysis via the arachidonate 5-lipoxygenase (ALOX5) /nuclear factor kappa B (NF-κB) signaling pathway on the progression of diabetic nephropathy (DN) . @*Methods @#Bioinformatics analysis was used to identify the target genes of SFN in the treatment of DN . Human proximal tubular epithelial cell line (HK-2 cells) was induced with 30 mmol/L high glucose (HG) to create an in vitro model of DN . HK-2 cells were divided into the following groups : normal glucose (NG) group , HG group , HG + SFN (3 mmol/L) group , HG + ALOX5 group , HG + SFN (3 mmol/L) + ALOX5 group , HG-treated macrophages + HK-2 group , HG + SFN (3 mmol/L) treated macrophages s + HK-2 group , HG + ALOX5 transfection treated macrophages + HK-2 group , HG + SFN (3 mmol/L) + ALOX5 transfection treated macrophages + HK-2 group . CCK-8 assay was used to detect cell viability , Terminal deoxynucleotidyl transferase- mediated dUTP nick-end labeling (TUNEL) method was used to detect cell apoptosis; glucose and lactate levels in the cells were measured using assay kits; Western blot was performed to detect the expression of ALOX5 , NF-κB , and glycolysis-related proteins hexokinase-2 ( HK2 ) , pyruvate kinase M2 ( PKM2 ) , glucose transporter 1 (GLUT1) in each group . Diabetic nephropathy (DN) mouse models were established using streptozotocin (STZ) and treated with SFN (0. 5 mg/kg) . Various biochemical parameters were measured in the mice , and kidney tissue pathology was examined using H&E staining. Western blot was used to detect the expression of glycolysis-related proteins (HK2 , PKM2 , GLUT1) in kidney macrophages . @*Results @# Bioinformatics analysis revealed ALOX5 as the target gene of SFN in treating DN . Compared to the HG group , SFN treatment enhanced HK-2 cell viability and in- hibited apoptosis (P < 0. 05) ; concurrently , SFN treatment suppressed HG-induced macrophage glycolysis-related protein and attenuated macrophage-mediated HK-2 cellular injury ( P < 0. 05) . Western blot results showed that SFN inhibited the expression of ALOX5 and NF-κB ( P < 0. 05) . The mouse experiment results showed that SFN treatment improved kidney function and pathological changes in the kidney of DN mice , and inhibited the related protein expression of acrophage glycolysis in kidney tissue (P < 0. 05) . @*Conclusion @#SFN improves the progression of DN by inhibiting the expression of macrophage glycolysis-related protein through the ALOX5/NF-κB signaling pathway .

Blumea balsamifera （L.） DC Oil Alleviates LPS-Induced Inflammatory Response in Macrophages by Affecting Arachidonic Acid Metabolism Via NF-κB Nonclassical Pathway / 中山大学学报(医学科学版)

ObjectiveTo study the anti-inflammatory effects of Blumea balsamifera （L.） DC oil （BBO） based on nuclear factor kappa-B （NF-κB） nonclassical and arachidonic acid （AA） pathway. MethodsEffects of BBO on the production of slow reacting substance of anaphylaxis （SRS-A） were detected by the ileal smooth muscle method. The contents of prostaglandin E2 （PGE2） and leukotriene B4 （LTB4） in lipopolysaccharides （LPS） -induced macrophages were detected by ELISA kit. The expression of COX-2， 5-LOX， FLAP and RelB were detected by qRT-PCR. Western blot was performed to detect the effects of BBO on the level of NF-κB nonclassical pathway proteins TNF receptor associated factor 3 （TRAF3）， TNF receptor associated factor 2 （TRAF2）， NF-κB-inducing kinase （NIK）， p100 and RelB. ResultsThe contractile tension of guinea pig ileum was reduced （P<0.001）， and the SRS-A production inhibition rate reached 65.34% at 1mg·mL-1 BBO concentration. Compared with LPS group， BBO reduced the concentrations of PGE2 （P<0.05） and LTB4 （P<0.05）， and decreased the expressions of COX-2 （P<0.05）， 5-LOX （P<0.05） and FLAP （P<0.05） in AA pathway at concentrations of 40-80 μg·mL-1. Moreover， 40-80 μg·mL-1 BBO decreased the concentrations of TRAF3 （P<0.05）， TRAF2 （P<0.05）， and NIK （P<0.05）， and further inhibited the phosphorylation of p100 （P<0.05）， as well as the level of the transcription factor RelB in genes （P<0.05） and proteins （P<0.05） in nonclassical NF-κB pathway， whereas BBO did not cause such changes. ConclusionBBO may potentially exert its anti-inflammatory effects by suppressing the regulatory proteins TRAF3 and TRAF2 and the transcription factor RelB in NF-κB nonclassical pathway. The inhibitory action extending to the induction kinase function of NIK， further hindering the phosphorylation of p100 and its binding with the transcription factor RelB. Consequently， downstream elements in the AA pathway， including the pivotal rate-limiting enzymes COX-2， 5-LOX and FLAP， were altered. This modulation influences the levels of inflammatory mediators such as PGE2 and LTB4.

Correlación entre la adicción a la comida y niveles del ácido araquidónico en adultos con obesidad tras intervenciones nutricionales / Correlation between food addiction and arachidonic acid levels in obese adults following nutritional interventions

Introducción: Los endocannabinoides son una diana en el tratamiento de la obesidad y se producen a partir de ácidos grasos esenciales, los derivados del ácido linoleico actúan como agonistas de los receptores cannabinoides tipo 1 (CB1), asimismo, los derivados del ácido linolénico ejercen efectos de antagonistas de dichos receptores, por lo cual se plantea que modificar el consumo dietario de los ácidos grasos omega 3 y 6 podría modular la activación del sistema endocannabinoide, lo que podría ser favorable para personas con adicción a la comida, considerando cómo este sistema promueve la actividad de las vías dopaminérgicas que se alteran en la adicción a sustancias psicoactivas. Objetivo: Analizar la correlación entre el puntaje de adicción a la comida por la escala mYFAS 2.0 y los niveles plasmáticos de ácido araquidónico en adultos con obesidad tras modular la ingesta de alimentos fuente de ácidos grasos esenciales. Metodología: Se desarrolló un estudio piloto con diseño de ensayo clínico cruzado en dos tiempos, en donde los participantes recibieron los tratamientos estándar y experimental, en estos se brindaron planes siguiendo recomendaciones para el manejo nutricional de la obesidad, adicionalmente, el tratamiento experimental contó con pautas para disminuir el consumo del Omega 6 y aumentar el consumo de Omega 3 para obtener una relación menor a 5:1 entre estos ácidos grasos. Resultados: Se observó una disminución significativa en el puntaje de adicción a la comida y los niveles plasmáticos de ácido araquidónico en los participantes tras recibir el tratamiento experimental, presentando una correlación directamente proporcional entre estas, por otro lado, el tratamiento estándar estuvo asociado a una correlación inversamente proporcional entre estos. Conclusiones: El descenso en las concentraciones plasmáticas del ácido araquidónico fue asociado a un menor puntaje en la escala mYFAS 2.0 de adicción a la comida en los participantes de este estudio tras su exposición al tratamiento experimental.

Introduction: Endocannabinoids are a target in obesity treatment and they are produced from the essential fatty acids, the metabolites of linoleic acid act as agonists of the cannabinoid receptors type 1 (CB1), likewise, the metabolites of the linolenic acid act as inverse agonists of such receptors, hence, it is proposed that modifying the dietary intake of the essential fatty acids (Omega 6 and 3) may modulate the activation of the endocannabinoid system, this could be favorable for people with food addiction, considering how this system promotes the activity of the dopaminergic pathways that are altered in the psychoactive substances addiction. Objective: To analyze the correlation between the food addiction score and plasmatic levels of arachidonic acid in adults with obesity following a modulation of the dietary intake of essential fatty acids n-6 and n-3 food sources. Methods: A pilot study was carried out with a two-period crossover clinical trial design, in which the participants received standard and experimental treatments, in these programs, plans were provided following guidelines for the nutritional management of obesity, in addition, the experimental treatment included recommendations to reduce the intake of linoleic acid and to increase the intake of linolenic acid to obtain a ratio lower to 5:1 between these fatty acids. Results: A significant decrease in the food addiction score and plasmatic levels of arachidonic acid was observed in the participants exposed to the experimental treatment, showing a directly proportional correlation, moreover, the standard treatment was associated to inverse correlations between these variables. Conclusion: The decrease in plasmatic arachidonic acid levels was associated with lower scores on the mYFAS 2.0 of food addiction in the participants of this study following their exposure to the experimental treatment.

Allosteric modulation on specific gene mutated Vitamin D receptor by essential PUFAs and its distinct molecular effects on type 2 diabetes mellitus

A serious health threat affecting the T2DM group is evident more cases T2DM are diagnosed. In this research, we choose to research into all of this possible mechanism of 3T3-L1 Cell lines and Molecular Docking studies Schrodinger software identified Vitamin D, Omega-3, and 6 PUFAs (EPA DHA & AA) Compounds of hydrophilic and hydrophobic pocket throughout molecular modeling besides T2DM. A group of three analog VDRs is being developed for discovery treatment with T2DM. Its use as it was agreed to run a molecular cell culture and docking study. Recognize the binding method involving the compound in T2DM through ADME prediction. The molecular dynamics simulation was enhanced by confirmation of the strength of the possible composite binding. Based on the computational results, the Omega-3 and 6 PUFAs compound encourages energy interaction. The composite contains an in vitro anti-diabetic activity; the compounds have clearly shown that they are active on T2DM. Our studies provide vital information on the findings of the bimolecular T2DM inhibitors.

Effects of Total Saponins from Dioscorea Nipponica Makino on Monosodium Urate-Induced M1-Polarized Macrophages through Arachidonic Acid Signaling Pathway: An in vitro Study / 中国结合医学杂志

OBJECTIVE@#To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino (TSDN) on the arachidonic acid pathway in monosodium urate (MSU) crystal-induced M1-polarized macrophages.@*METHODS@#M1 polarization of RAW264.7 cells were induced by 1 µ g/mL lipopolysaccharide (LPS). The methylthiazolyldiphenyl-tetrazolium bromide method was then used to screen the concentration of TSDN. MSU (500 µ g/mL) was used to induce the gouty arthritis model. Afterwards, 10 µ g/L TSDN and 8 µ mol/L celecoxib, which was used as a positive control, were added to the above LPS and MSU-induced cells for 24 h. The mRNA and protein expressions of cyclooxygenase (COX) 2, 5-lipoxygenase (5-LOX), microsomal prostaglandin E synthase derived eicosanoids (mPGES)-1, leukotriene B (LTB)4, cytochrome P450 (CYP) 4A, and prostaglandin E2 (PGE2) were tested by real-time polymerase chain reaction and Western blotting, respectively. The enzyme-linked immunosorbent assay was used to test the contents of M1 markers, including inducible nitric oxid synthase (NOS) 2, CD80, and CD86.@*RESULTS@#TSDN inhibited the proliferation of M1 macrophages and decreased both the mRNA and protein expressions of COX2, 5-LOX, CYP4A, LTB4, and PGE2 (P<0.01) while increased the mRNA and protein expression of mPGES-1 (P<0.05 or P<0.01). TSDN could also significantly decrease the contents of NOS2, CD80, and CD86 (P<0.01).@*CONCLUSION@#TSDN has an anti-inflammation effect on gouty arthritis in an in vitro model by regulating arachidonic acid signaling pathway.

Mechanism of tryptanthrin in treatment of ulcerative colitis in mice based on serum metabolomics / 中国中药杂志

This study aims to explore the effect of tryptanthrin on potential metabolic biomarkers in the serum of mice with ulcerative colitis(UC) induced by dextran sulfate sodium(DSS) based on liquid chromatography-mass spectrometry(LC-MS) and predict the related metabolic pathways. C57BL/6 mice were randomly assigned into a tryptanthrin group, a sulfasalazine group, a control group, and a model group. The mouse model of UC was established by free drinking of 3% DSS solution for 11 days, and corresponding drugs were adminsitrated at the same time. The signs of mice were observed and the disease activity index(DAI) score was recorded from the first day. Colon tissue samples were collected after the experiment and observed by hematoxylin-eosin(HE) staining. The levels of interleukin-4(IL-4), interleukin-10(IL-10), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-8(IL-8) in the serum were measured by enzyme linked immunosorbent assay(ELISA). The serum samples were collected from 6 mice in each group for widely targeted metabolomics. The metabolic pathways were enriched by MetaboAnalyst 5.0. The results showed that compared with the model group, tryptanthrin treatment decreased the DAI score(P<0.05), alleviated the injury of the colon tissue and the infiltration of inflammatory cells, lowered the levels of proinflammatory cytokines, and elevated the levels of anti-inflammatory cytokines in the serum. The metabolomic analysis revealed 28 differential metabolites which were involved in 3 metabolic pathways including purine metabolism, arachidonic acid metabolism, and tryptophan metabolism. Tryptanthrin may restore the metabolism of the mice with UC induced by DSS to the normal level by regulating the purine metabolism, arachidonic acid metabolism, and tryptophan metabolism. This study employed metabolomics to analyze the mechanism of tryptanthrin in the treatment of UC, providing an experimental basis for the utilization and development of tryptanthrin.

Total Saponin Fraction of Dioscorea Nipponica Makino Improves Gouty Arthritis Symptoms in Rats via M1/M2 Polarization of Monocytes and Macrophages Mediated by Arachidonic Acid Signaling / 中国结合医学杂志

OBJECTIVE@#To explore the mechanism of effects of total saponin fraction from Dioscorea Nipponica Makino (TSDN) on M1/M2 polarization of monocytes/macrophages and arachidonic acid (AA) pathway in rats with gouty arthritis (GA).@*METHODS@#Seventy-two Sprague Dawley rats were randomly divided into 4 groups (n=18 in each): normal, model, TSDN at 160 mg/kg, and celecoxib at 43.3 mg/kg. Monosodium urate crystal (MSU) was injected into the rats' ankle joints to induce an experimental GA model. Blood and tissue samples were collected on the 3rd, 5th, and 8th days of drug administration. Histopathological changes in the synovium of joints were observed via hematoxylin and eosin (HE) staining. The expression levels of arachidonic acid (AA) signaling pathway were assessed via real-time polymerase chain reaction (qPCR) and Western blot. Flow cytometry was used to determine the proportion of M1 and M2 macrophages in the peripheral blood. An enzyme-linked immunosorbent assay (ELISA) was used to detect interleukine (IL)-1 β, tumor necrosis factor-alpha (TNF-α), IL-4, IL-10, prostaglandin E2 (PGE2), and leukotriene B4 (LTB4).@*RESULTS@#HE staining showed that TSDN improved the synovial tissue. qPCR and Western blot showed that on the 3rd, 5th and 8th days of drug administration, TSDN reduced the mRNA and protein expressions of cyclooxygenase (COX)2, microsomal prostaglandin E synthase-1 derived eicosanoids (mPGES-1), 5-lipoxygenase (5-LOX), recombinant human mothers against decapentaplegic homolog 3 (Smad3), nucleotide-binding oligomerization domain-like receptor protein 3 (NALP3), and inducible nitric oxide synthase (iNOS) in rats' ankle synovial tissues (P<0.01). TSDN decreased COX1 mRNA and protein expression on 3rd and 5th day of drug administration and raised it on the 8th day (both P<0.01). It lowered CD68 protein expression on days 3 (P<0.01), as well as mRNA and protein expression on days 5 and 8 (P<0.01). On the 3rd, 5th, and 8th days of drug administration, TSDN elevated the mRNA and protein expression of Arg1 and CD163 (P<0.01). Flow cytometry results showed that TSDN decreased the percentage of M1 macrophages while increasing the percentage of M2 in peripheral blood (P<0.05 or P<0.01). ELISA results showed that on the 3rd, 5th, and 8th days of drug administration, TSDN decreased serum levels of IL-1 β, TNF-α, and LTB4 (P<0.01), as well as PGE2 levels on days 3rd and 8th days (P<0.05 or P<0.01); on day 8 of administration, TSDN increased IL-4 serum levels and enhanced IL-10 contents on days 5 and 8 (P<0.05 or P<0.01).@*CONCLUSION@#The anti-inflammatory effect of TSDN on rats with GA may be achieved by influencing M1/M2 polarization through AA signaling pathway.

Metabolomics study of rat tail vibration model / 环境与职业医学

Background The metabolites and metabolic pathways of hand-arm vibration syndrome have not yet been elucidated. Objective To investigate the effect of local vibration on endogenous metabolites in rat serum by metabolomic analysis, to preliminarily explore the potential metabolic pathway of endogenous metabolites, so as to provide evidence for further research on the mechanism of hand-arm vibration syndrome. Methods Thirty-two SPF male SD rats, (211.3±11.1) g, 7−8 weeks of age, were selected and randomly divided into three groups: control group (14 rats, without vibration), 7 d vibration group (9 rats, continuously vibration for 7 d), and 14 d vibration group (9 rats, continuous vibration for 14 d). The vibration rats were vibrated every day for 4 h, the frequency weighted acceleration was 4.9 m·s−2, the vibration frequency was 125 Hz, and the vibration direction was one-way vertical vibration. The control group had the same conditions except not contacting vibration. After the vibration exposure, the blood samples taken from the abnormal aorta of rats were collected, and the changes of rat serum metabolome were analyzed by ultra-performance liquid chromatography-tandem time-of-flight mass spectrometry. Principal components analysis (PCA) was used to explore changes in rat serum metabolic profile, and orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to screen out differential metabolites. Combined with online databases, a metabolic pathway enrichment analysis of differential metabolites was performed. Results The PCA analysis showed that compared with the control group, the rat serum metabolic profiles in the 7 d group and the 14 d group were clearly differentiated, and the rat serum metabolic profiles in the 7 d group and the 14 d group partially overlapped. The OPLS-DA analysis showed significant differences between groups. The main parameters were: model interpretation rate R2Y=0.914, model predictive ability Q2=0.58. The OPLS-DA analysis screened out 26 and 119 differential metabolites from the 7 d group and the 14 d group respectively, and there were 24 common differential metabolites between the 7 d group and the 14 d group. The metabolomic pathway analysis showed that local vibration-induced changes in rat serum metabolism were mainly related to arachidonic acid metabolism in the 14 d group, among which the metabolites with significant effects were arachidonic acid, prostaglandin E2, and prostaglandin D2. Conclusion Local vibration could affect the normal metabolism in rats, and the metabolic pathway with significant influence is arachidonic acid metabolism after a 14 d exposure and the involved metabolites are arachidonic acid, prostaglandin E2, and prostaglandin D2.

Based on plasma metabonomics and network analysis to research the mechanisms of Chaigui granules for treating depression / 药学学报

The purpose of this study was to systematically analyze the antidepressant mechanism of Chaigui granules from the perspective of biological metabolic network by using integrated metabolomics and biological network analysis tools. The model of chronic unpredictable mild stress (CUMS) depression rat was established, and LC-MS-based plasma metabolomics was used to identify the key metabolites and analyze metabolic pathways underlying the antidepressant effects of Chaigui Granules. The key metabolites regulated by Chaigui granules was integrated with biological network analysis tools to further focus on the key metabolic pathways and explore the potential targets of the antidepressant effect of Chaigui granules. The results showed that there were significant differences in the plasma levels of 20 metabolites in the model group compared with the control group (P < 0.05), Chaigui granules significantly regulated 12 metabolites including docosatrienoic acid, 3-hydroxybutyric acid, 4-hydroxybenzaldehyde, chenodeoxycholic acid, cholic acid, L-glutamine, glycocholic acid, linoleyl carnitine, L-tyrosine, N-acetylvaline, palmitoylcarnitine, arachidonic acid. Further network analysis of the key metabolites regulated by Chaigui granules indicated that plasma arachidonic acid metabolism might be the core pathway for the antidepressant effect of Chaigui granules, with 10 proteins were potential targets for the antidepressant effect of Chaigui granules, including CYP2B6, CYP2E1, CYP2C9, CYP2C8, PLA2G6, PTGS2, ALOX15B, PTGS1, ALOX12 and ALOX5. The animal experimental operations involved in this paper was followed the regulations of the Animal Ethics Committee of Shanxi University and passed the animal experimental ethical review (Approval No. SXULL2020028).

Mechanism of omege-6 PUFA increasing the susceptibility to acute myocardial ischemia injury / 药学学报

The abnormal lipids metabolism is a critical pathological feature of coronary heart disease (CHD). Additional supplemental intake of polyunsaturated fatty acid (PUFA) has long been considered to be an effective strategy for preventing CHD, but more and more clinical trials have denied this view. Still, it is ambiguity for the specific mechanism of PUFA in CHD. The experimental programs are compliant with ethical principles for animal use and have been approved by the Animal Experiment Ethics Committee of Jinan University. In the present study, we established an animal model by intake of omega-6 PUFA combined acute myocardial ischemia to explore the mechanism of CHD. Intragastric administration of linoleic acid (LA) for 14 days, intraperitoneal injection of isoprenaline (ISO) was applied to induce acute myocardial ischemia for the animal model establishment. The animal ultrasound imaging system was used to detect cardiac function in vivo after ISO injection for 24 h. Serum and heart tissue samples were collected for the myocardial enzyme, phospholipidomics analysis and molecular biological detection. Compared to the LA group, the cardiac function showed that the left ventricular ejection fraction (EF%) and the left ventricular shortening fraction (FS%) decreased, aspaetate aminotransferase (AST), creatine kinase isoenzyme (CK-MB), and lactate dehydrogenase (LDH) increased in the LA + ISO mice. Compared to the ISO group, the phospholipidomics analysis showed that the PUFAs significantly were raised in the LA + ISO myocardium, and the content of oxidized phosphatidylethanolamine (ox-PE) changed most remarkable. Compared with the ISO group, the molecular biology detection showed that glutathione (GSH) and nicotinamide adenine dinucleotide phosphate (NADPH) were depleted, the end-products of ox-PE were increased, and the level of arachidonic acid 12/15-lipoxygenase (ALOX15) protein expression increased obviously. We suggest that ALOX15 mediated phospholipid peroxidation might be the critical mechanism of LA increased the susceptibility of myocardial ischemia injury. This study provides an experimental basis for whether PUFA could be used as an alternative treatment strategy for CHD prevention and provides a new intervention target for the early prevention strategy of CHD.

Effects of ophiopogonin D on fatty acid metabolic enzymes in cardiomyocytes / 中国中药杂志

To explore the effect of ophiopogonin D on main fatty acid metabolic enzymes in human cardiomyocyte AC-16,so as to provide reference for cardiovascular protection mechanism and safe clinical application of Ophiopogon japonicus.CCK-8 (cell counting kit-8) was used to detect the effect of different concentrations of ophiopogonin D on the viability of cardiomyocytes.Meanwhile,the effect of different concentrations of ophiopogonin D on the morphology and quantity of cardiomyocytes was observed under microscope.The effect of ophiopogonin D on the mRNA expression of CYP2J2,CYP4F3,CYP4A11,CYP4A22 and CYP4F2 in cardiomyocytes was detected by RT-PCR.Western blot was used to detect the protein expression of CYP4F3 in different concentrations of ophiopogonin D.Compared with the control group,low-concentration ophiopogonin D had no effect on the viability of cardiomyocytes.However,ophiopogonin D with a concentration of higher than 20μmol·L~(-1)could promote the viability.Under the microscope,ophiopogonin D with a concentration of below 100μmol·L~(-1)had no significant effect on the morphology and number of cardiomyocytes.RT-PCR results showed that compared with the control group,5μmol·L~(-1)ophiopogonin D could slightly up-regulate mRNA expressions of CYP2J2 and CYP4F3,while high-concentration ophiopogonin D (10 and 20μmol·L~(-1)) could significantly induce mRNA expressions of CYP2J2and CYP4F3 in a dose-dependent manner (P<0.05).The same concentration of ophiopogonin D had a little effect on the mRNA expressions of CYP4A11,CYP4A22 and CYP4F2.Western blot results showed that 20μmol·L~(-1)ophiopogonin D could significantly induce the protein expression of CYP4F3 in a dose-dependent manner (P<0.05).Based on the above results,ophiopogonin D (less than100μmol·L~(-1)) has no effect on the viability of AC-16 cardiomyocytes.Ophiopogonin D (less than 100μmol·L~(-1)) can selectively induce the expressions of CYP2J2 and CYP4F3,regulate the metabolic pathway of fatty acid signaling molecules,and thus protecting the cardiovascular system.

Development of a highly-specific

As a serine hydrolase, monoacylglycerol lipase (MAGL) is principally responsible for the metabolism of 2-arachidonoylglycerol (2-AG) in the central nervous system (CNS), leading to the formation of arachidonic acid (AA). Dysfunction of MAGL has been associated with multiple CNS disorders and symptoms, including neuroinflammation, cognitive impairment, epileptogenesis, nociception and neurodegenerative diseases. Inhibition of MAGL provides a promising therapeutic direction for the treatment of these conditions, and a MAGL positron emission tomography (PET) probe would greatly facilitate preclinical and clinical development of MAGL inhibitors. Herein, we design and synthesize a small library of fluoropyridyl-containing MAGL inhibitor candidates. Pharmacological evaluation of these candidates by activity-based protein profiling identified

Clinical significance of EPHX2 deregulation in prostate cancer / 亚洲男科学杂志(英文版)

The arachidonic acid (AA) metabolic pathway participates in various physiological processes as well as in the development of malignancies. We analyzed genomic alterations in AA metabolic enzymes in the Cancer Genome Atlas (TCGA) prostate cancer (PCa) dataset and found that the gene encoding soluble epoxide hydrolase (EPHX2) is frequently deleted in PCa. EPHX2 mRNA and protein expression in PCa was examined in multiple datasets by differential gene expression analysis and in a tissue microarray by immunohistochemistry. The expression data were analyzed in conjunction with clinicopathological variables. Both the mRNA and protein expression levels of EPHX2 were significantly decreased in tumors compared with normal prostate tissues and were inversely correlated with the Gleason grade and disease-free survival time. Furthermore, EPHX2 mRNA expression was significantly decreased in metastatic and recurrent PCa compared with localized and primary PCa, respectively. In addition, EPHX2 protein expression correlated negatively with Ki67 expression. In conclusion, EPHX2 deregulation is significantly correlated with the clinical characteristics of PCa progression and may serve as a prognostic marker for PCa.

Exploration on molecular mechanism of hot herb Aconitum carmichaelii for therapeutic action based on metabolomics / 中草药

Objective: To observe the effects of Aconitum carmichaelii on serum metabolites in mice by metabolomics technology, and to explore biomarkers and metabolic pathways and targets related to its treatment of various ailments such as cardiovascular diseases, in order to uncover its molecular mechanism of efficacy. Methods: Twenty male mice were randomly divided into two groups, and A. carmichaelii decoction and distilled water were orally administered with the dose of 15 mL/(kg∙d) for consecutive 4 d respectively. Collected blood samples of each group were analyzed using UPLC-MS/MS technology, and data pattern recognition was performed using PCA, PLS-DA and other analytical methods. Meanwhile, differential metabolites were screened out based on VIP greater than 1 and manual integral calculation. The differential metabolites were used for pathway analysis. Network modular analysis and targets screening were performed by Cytoscape and MetScape. Results: When performing data pattern recognition, the aconite group and the control group could be completely separated. A total of 18 differential metabolites were screened out, and their contents were up-regulated. Pathway analysis was performed to obtain five related pathways, namely linoleic acid metabolism, arachidonic acid metabolism, starch and sucrose metabolism, nicotinate and nicotinamide metabolism, and inositol phosphate metabolism. Fourteen modules were obtained using the network analysis, the largest two of which were arachidonic acid metabolism pathway and linoleic acid metabolism pathway. The degree of arachidonic acid (59), linoleic acid (55), nicotinamide (26), and palmitic acid (11) were greater than the mean value (8.010) in the network, and the related pathways were arachidonic acid metabolism, linoleic acid metabolism, nicotinate and nicotinamide metabolism, and saturated fatty acid beta-oxidation pathway respectively. A total of 26 genes were screened out, all of which belonged to the cytochrome P450 enzyme system. Conclusion: A. carmichaelii may affect arachidonic acid metabolism by acting on CYP450, thereby improving the body's energy metabolism and producing therapeutic effects.

Study on network pharmacology mechanism of Leonurus japonicus in treating dysmenorrhea caused by cold coagulation and blood stasis / 中草药

Objective: To establish a chemical constitution-disease target-metabolic signaling pathway network of Leonurus japonicus, and explore the mechanism of multi-component, multi-target and multi-pathway of L. japonicus for the treatment of dysmenorrhea caused by cold coagulation and blood stasis. Methods: TCMSP database and Swiss Target Prediction server were used to obtain the chemical components and action targets of L. japonicus. Combined with DrugBank, DisGeNET, TTD and other databases, the action targets of L. japonicus for treating dysmenorrhea symptoms wtih cold coagulation and blood stasis were obtained. Through DAVID's website, GO function enrichment analysis and KEGG pathway analysis were conducted on the targets, and cytoscape 3.6.0 was used to construct the network diagram of the active component-dysmenorrhea target-metabolic signaling pathway of L. japonicus, to explore the mechanism of action of L. japonicus in treating dysmenorrhea with cold coagulation and blood stasis. Furthermore, Western blotting experiments were conducted to verify the effect of L. japonicus on the expression of PTGS1 and PTGS2 proteins in the uterine tissues of the model rats with cold coagulation and blood stasis dysmenorrhea. Results: Eight potential active ingredients of L. japonicus were obtained, including 22 dysmenorrhea related disease targets, main targets PTGS1, PTGS2, ALOX5, PLAG2A, AKR1C3, etc. A total of 71 GO items were obtained by GO functional enrichment analysis (P < 0.05), including 46 biological process (BP) items, four cell component (CC) items, and 21 molecular function (MF) items. There were 22 possible targets for the treatment of dysmenorrhea, and seven signaling pathways were obtained through KEGG pathway enrichment and screening (P < 0.05), involving arachidonic acid metabolism, linoleic acid metabolism, steroid hormone biosynthesis, etc. L. japonicus significantly increased the protein expression levels of PTGS1 and PTGS2 in the uterus of the model rats (P < 0.05), which confirmed some of the predicted results of network pharmacology. Conclusion: L. japonicus may treat dysmenorrhea with cold coagulation and blood stasis by acting on arachidonic acid metabolism pathway.

Study on treatment of "cytokine storm" by anti-2019-nCoV prescriptions based on arachidonic acid metabolic pathway / 中国中药杂志

Since the outbreak of 2019-nCoV, the epidemic has developed rapidly and the situation is grim. LANCET figured out that the 2019-nCoV is closely related to "cytokine storm". "Cytokine storm" is an excessive immune response of the body to external stimuli such as viruses and bacteria. As the virus attacking the body, it stimulates the secretion of a large number of inflammatory factors: interleukin(IL), interferon(IFN), C-X-C motif chemokine(CXCL) and so on, which lead to cytokine cascade reaction. With the exudation of inflammatory factors, cytokines increase abnormally in tissues and organs, interfering with the immune system, causing excessive immune response of the body, resulting in diffuse damage of lung cells, pulmonary fibrosis, and multiple organ damage, even death. Arachidonic acid(AA) metabolic pathway is principally used to synthesize inflammatory cytokines, such as monocyte chemotactic protein 1(MCP-1), tumor necrosis factor(TNF), IL, IFN, etc., which is closely related to the occurrence, development and regression of inflammation. Therefore, the inhibition of AA metabolism pathway is benefit for inhibiting the release of inflammatory factors in the body and alleviating the "cytokine storm". Based on the pharmacophore models of the targets on AA metabolic pathway, the traditional Chinese medicine database 2009(TCMD 2009) was screened. The potential herbs were ranked by the number of hit molecules, which were scored by pharmacophore fit value. In the end, we obtained the potential active prescriptions on "cytokine storm" according to the potential herbs in the "National novel coronavirus pneumonia diagnosis and treatment plan(trial version sixth)". The results showed that the hit components with the inhibitory effect on AA were magnolignan Ⅰ, lonicerin and physcion-8-O-β-D-glucopy-ranoside, which mostly extracted from Magnoliae Officinalis Cortex, Zingiberis Rhizoma Recens, Lonicerae Japonicae Flos, Rhei Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma, Scutellariae Radix, Gardeniae Fructus, Ginseng Radix et Rhizoma, Arctii Fructus, Dryopteridis Crassirhizomatis Rhizoma, Paeoniaeradix Rubra, Dioscoreae Rhizoma. Finally the anti-2019-nCoV prescriptions were analyzed to obtain the potential active prescriptions on AA metabolic pathway, Huoxiang Zhengqi Capsules, Jinhua Qinggan Granules, Lianhua Qingwen Capsules, Qingfei Paidu Decoction, Xuebijing Injection, Reduning Injection and Tanreqing Injection were found that may prevent 2019-nCoV via regulate cytokines. This study intends to provide reference for clinical use of traditional Chinese medicine to resist new coronavirus.

Recent advance in epoxyeicosatrienoic acids in intracerebral hemorrhage / 中华神经医学杂志

Epoxyeicosatrienoic acids (EETs) are mainly from intracellular arachidonic acids catalyzed by cytochrome P450 cyctooxygenase and degraded to lower active dihydroxyeicosapentaenoic acids by soluble epoxide hydrolase.In recent years,EETs have been found to be a new target for prevention and treatment of various nervous system diseases,such as anti-inflammatory reaction,anti-atherosclerosis,anti-cell apoptosis and angiogenesis.Intracerebral hemorrhage is a kind of serious acute cerebrovascular disease.Cerebral hemorrhage is a kind of acute cerebrovascular disease;secondary injury is one of the important mechanisms of cerebral hemorrhage;the present studies have confirmed that EETs have protective role in brain tissues after cerebral hemorrhage,thus,become new hotspot in the research of cerebral hemorrhage.This review focuses on the role and mechanism of EETs in intracerebral hemorrhage,hoping to provide some references for exploration of new research directions and therapeutic targets in the treatment ofintracerebral hemorrhage.

Effect of Oral Administration of Diclofenac Sodium on Coagulation Factors and Some Hematological Parameters in Wister Rats

Diclofenac sodium is a nonsteroidal anti-inflammatory drug often obtainable as a prescription drug or over the counter. It is very effective in the control of inflammation and pain due to arthritis or pains arising following many disease conditions becauseof its antipyretic, anti inflammatory and analgesic potentials. Despite the beneficial effects of diclofenac sodium, it has been implicated in some adverse effects. In this study, we examined the effect of acute and chronic administration of diclofenac sodium on some hematological (PCV, WBC differentials) and coagulation (prothrombin time, activated partial prothrombin time and platelets count) parameters of albino Wister rats using the standard methods. Twenty four Albino Wister rats were divided intothree groups of 8 rats and grouped as control, acute study and chronic study. The rats were administered 0.2 mg of diclofenac sodium for 24 hours for acute and 3 weeks for chronic studies respectively. The rats were sacrificed and blood collected for analysis of PCV, WBC differentials, prothrombin time, activated partial prothrombin time and platelets count using the standard methods. Results show that acute administration of diclofenac sodium at 0.2 mg has no effect on hematological and coagulation parameters, but chronic administration could instigate significant reduction in PCV, platelets count, neutrophils and monocytes (p<0.001), whilethere is a significant increase in PT, INR, lymphocytes (p<0.001). Considering these alterations, it is advisable that this drug should be made a strictly prescription drug in order to prevent indiscriminate use of this medication and to prevent attendant anemia and coagulopathy that may follow chronic use.

Assessment Of Analgesic Activity Of Nelumbo Nucifera Fruit Ethanol Extract

Objective: Utilization of herbal remedies rich in flavonoids and vitamins have increased significantly these days to treat various disorders, thus existing research work encircled to appraise the analgesic effect of Nelumbo nucifera fruit (NNF) for evaluating its traditional use pharmacologically in disorders which are associated with pain and inflammation. Methods: Central analgesic activity in mice was assessed by tail flick test and the latency time i.e. the removal of tail from the stimulus was recorded. Similarly acetic acid induced writhing test was also conducted for the assessment of peripheral analgesic effect in mice and number of writhes was counted along with percent inhibition of writhes. Results: In tail flick test the peek anti-nociceptive effect at all doses of fruit was observed at 90 min. However, the percentage of tail elongation time was highest at a dose of 200 mg/kg i.e. 82% at 90 min. Number of writhes was highly significantly reduced at all doses of NNF but maximum effects were observed at dose 200 mg/kg as compared to control, indicating 48.41 % inhibition of writhes. Conclusion: NNF have exhibited strong analgesic effect in both animal models, which may be connected with the synergistic actions of flavonoids, saponins and tannins on arachidonic acid pathway inhibition. Hence NNF seems to have a great potential in disorders associated with pain but more experimental trials in this field are required to confirm these findings.

Pharmacological research progress of sodium ferulate in cardio-cerebral vascular disease / 中草药

Sodium ferulate is the sodium salt of ferulic acid, an extract of traditional Chinese herbal medicines for promoting blood circulation and detoxification, and it is rich in sources, with few side effects and high safety. Sodium ferulate has many pharmacological effects, which is a cardiovascular drug researched and developed independently in China. It was first approved in 1990 for the clinical treatment of cardiovascular diseases. In recent years, the clinical application of sodium ferulate has become increasingly widespread, and the research field is continuously expanding. Sodium ferulate is effective in treating respiratory diseases, diabetes and complications, and protecting the liver and kidney from damage. Meanwhile it has been widely used in cardiovascular diseases. Here we reviewed the research status of the prominent pharmacological effects of sodium ferulate on cardiovascular diseases in the past 30 years, mainly focusing on the antithrombotic effects, the protection of blood vessels, and the anti-oxidative effect of sodium ferulate. It is expected to provide guidance for clinical applications of sodium ferulate.