OBJECTIVETo investigate the effects of CysLT receptor agonist leukotriene D4(LTD4) and antagonists on activation of microglia BV2 cells.

METHODSThe expression of CysLT1 and CysLT2 protein was determined by Western blotting and immunostaining in microglia BV2 cells. BV2 cells were pretreated with or without CysLT1 receptor selective antagonist montelukast, CysLT2 receptor selective antagonist HAMI 3379, or CysLT1/CysLT2 receptor dual antagonist BAY u9773 for 30 min, then the cells were treated with LTD4 for 24 h. Cell viability was detected by MTT reduction assay. Phagocytosis and mRNA expression of IL-6 were determined by fluorescent bead tracking and RT-PCR, respectively.

RESULTSIn BV2 cells, LTD4 did not affect proliferation but significantly enhanced phagocytosis and increased IL-6 mRNA expression in a concentration-dependent manner. LTD4 at 100 nmol/L induced a 1.4-fold increase of phagocytic index and a 2-fold up-regulation of IL-6 mRNA expression (P<0.01). HAMI 3379 and BAY u9773 (100 nmol/L) further increased LTD4-induced phagocytosis; BAY u9773 and montelukast decreased LTD4-induced IL-6 mRNA expression, while HAMI 3379 had no effect on that.

CONCLUSIONLTD4 activates BV2 cells in vitro and enhances IL-6 mRNA expression mediated by CysLT1 receptor, LTD4 induces phagocytosis which might be negatively regulated by CysLT2 receptor in BV2 cells.

Acetates ; pharmacology ; Cell Line ; Cell Proliferation ; Cyclohexanecarboxylic Acids ; pharmacology ; Humans ; Interleukin-6 ; metabolism ; Leukotriene Antagonists ; pharmacology ; Leukotriene D4 ; pharmacology ; Microglia ; cytology ; metabolism ; Phagocytosis ; Phthalic Acids ; pharmacology ; Quinolines ; pharmacology ; Receptors, Leukotriene ; metabolism ; SRS-A ; analogs & derivatives ; pharmacology

BACKGROUND AND OBJECTIVES: Both the granulation tissue and cholesterol granuloma can erode the surrounding bone and ossicles. However, the etiology of bone resorption in the granulation tissue and cholesterol granuloma has not been evident. The aim of this study was to assay the concentrations of arachidonic acid metabolites (AAMs) in cholesterol granuloma and the mastoid granulation tissue in order to better understand the possible role of AAMs in the pathophysiology of cholesterol granuloma in comparison with the granulation tissue. MATERIALS AND METHOD: Cholesterol granuloma tissues were obtained from eight patients who had underwent tympanomastoidectomy. Granulation tissues, which served for comparison, were taken from 12 patients who had underwent tympanomastoidectomy. Tissue concentrations of prostagladin (PG)E2, 6-keto-PGF1alpha, leukotriene (LT)C4, LTD4, LTE4, 15-hydro xyeicosatetraenoic acid (HETE), 12-HETE, 5-HETE, and thromboxane (TXB)2 were calculated using high performance liquid chromatography and compared between cholesterol granuloma and granulation tissue. RESULT: The level of 12-HETE was higher in cholesterol granuloma than in the granulation tissue. Among the PGs, the tissue concentration of PGE2 was particularly high in cholesterol granuloma than in the granulation tissue. LTD4 was the only LT detectable in cholesterol granuloma. In comparison to the AAMs in cholesterol granuloma, the lipoxygenase pathway products such as 12-HETE, 15-HETE, and 5-HETE were present in lower concentrations in the granulation tissue. LT was undetectable in the granulation tissues. CONCLUSION: Our results suggest that cholesterol granuloma and the granulation tissue is different not only in terms of histology, but also in terms of biochemical properties such as arachidonic acid metabolism.
12-Hydroxy-5,8,10,14-eicosatetraenoic Acid ; Arachidonic Acid* ; Arachidonic Acids ; Bone Resorption ; Cholesterol* ; Chromatography, Liquid ; Cytochrome P-450 CYP1A1 ; Dinoprostone ; Granulation Tissue* ; Granuloma* ; Humans ; Leukotriene D4 ; Leukotriene E4 ; Lipoxygenase ; Mastoid ; Metabolism ; Otitis Media* ; Otitis*

OBJECTIVE: Autistic spectrum traits are postulated to lie on a continuum that extends between individuals with autism and individuals with typical development. The present study was carried out to investigate functional and network abnormalities associated with autistic spectrum trait in healthy male subjects. METHODS: Subjects were 41 healthy male subjects who underwent the social responsiveness scale-adult (SRS-A) and magnetic resonance imaging. RESULTS: There was significant positive correlation between the total score of SRS-A and the regional cerebral blood flow (CBF) in posterior cingulate cortex (PCC). Also, there were changes in functional network such as in cingulate corti, insula and fusiform cortex. Further, we also found the significant difference of functional networks between the healthy male subjects with high or low autistic spectrum trait, and these points were congruent with the previous perceptions derived from autistic-spectrum disorders. CONCLUSION: These findings suggest a biological basis for the autistic spectrum trait and may be useful for the imaging marker of autism symptomatology.
Autism Spectrum Disorder ; Autistic Disorder ; Cerebrovascular Circulation* ; Gyrus Cinguli ; Humans ; Magnetic Resonance Imaging ; Male* ; SRS-A

That mast cells play a role in acute allergic inflammation by releasing various inflammatory mediators, including histamine, leukotrienes (LT), such as LTC4 and LTD4, and prostaglandins (PG), such as PGD2, is well known. Additionally, mast cells contribute to the development of allergic inflammation also through the release of multifunctional cytokines. The incidence of intraepithelial mast cells (IEMC) is found to be greater in nasal mucosa exposed to an allergen, and the cells are thought to play an important role in producing the immediate allergic reaction. Lamina propira mast cells (LPMC) are known to be the dominant source of TH2 cytokine and are responsible for development of the late phases of an allergic reaction They may upregulate the expression of adhesion molecules on the endothelial cells and induce basophil and eosinophil recruitment. Based on these consideration it can be proposed that mast cell is a initiating cell of allergic reaction in target organ and IEMC and LPMC have capacity to make major contribution to both immediate or late phase reaction of allergic rhinitis.
Basophils ; Cytokines ; Endothelial Cells ; Eosinophils ; Histamine ; Hypersensitivity ; Incidence ; Inflammation ; Leukotriene C4 ; Leukotriene D4 ; Leukotrienes ; Mast Cells* ; Nasal Mucosa ; Prostaglandin D2 ; Prostaglandins ; Rhinitis*

OBJECTIVETo study inhibitory the effects of Cryptoporus volvatus ferment substance(CVFS) on leukotriene production in vitro from neutrophils in rats.

METHODSNeutrophil aggregation was induced by intraperitoneal injection of glycogen in rats. After 16 h, intraperitoneal lavage fluid(PLF) was collected and neutrophils were removed. Neutrophils were stimulated by calcium ionophore A23187 in vitro to produce leukotriene B(4), C(4), D(4). The concentrations of leukotriene B(4), C(4) and D(4) were measured by reversed-phase high-performance liquid chromatography(HPLC).

RESULTCVFS at 0.25, 1, 4 mg x L(-1)decreased leukotriene B(4), C(4), D(4) release from neutrophils in a concentration-dependent manner. Inhibitory rate of CVFS 0.25, 1, 4 mg x L(-1 )on A23187-induced leukotriene B(4) production was 27.4%, 54.2% and 78.8%(P<0.05), respectively. Inhibitory rate of leukotriene C(4) production was 65.1%, 74.3 and 79.0%(P<0.05), respectively. Inhibitory rate of leukotriene D(4) production was 55.6%, 60.9% and 72.8%(P<0.05), respectively.

CONCLUSIONThe results suggest that suppression of leukotriene release may be a mechanism of the anti-inflammation and anti-asthma effects of CVFS.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Fermentation ; Leukotriene B4 ; secretion ; Leukotriene C4 ; secretion ; Leukotriene D4 ; secretion ; Male ; Neutrophils ; drug effects ; physiology ; Polyporaceae ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the changing contents of leukotriene B4 ( LTB4 ), leukotriene C4 ( LTC4 ), and leukotriene D4 (LTD4 ) of lung tissue in asthmatic rats, and explore the effect of Shuanglong Capsule (SLC) on it.

METHODSSD rats were randomly divided into the nomal group, asthmatic model group, Dexamethasone group and the high, middle and low dose SLC groups. All rats except those in the normal group were sensitized by ovalbumin and challenged with the antigen, and the contents of LTB4, LTC4 and LTD4 in lung tissue of all the groups were measured by reverse phase-high performance liquid chromatography (RP-HPLC) and compared.

RESULTSThe levels of LTB4, LTC4, and LTD4 of asthmatic rats were significantly higher than those of rats in the normal group. Dexamethasone and SLC at the dose of 8. 27 g/kg or 4. 13 g/kg could significantly inhibit the production of leukotrienes of lung tissue in asthmatic rats (P <0.05).

CONCLUSIONSLC can significantly inhibit the formation of inflammatory medium LTs of lung tissue in asthmatic rats, it may be one of the key mechanisms of SLC in anti-asthma and anti-inflammatory action.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Asthma ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Leukotriene B4 ; metabolism ; Leukotriene C4 ; metabolism ; Leukotriene D4 ; metabolism ; Leukotrienes ; metabolism ; Lung ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tablets

A reversed-phase high-performance liquid chromatography (RP-HPLC) has been developed to analyze the metabolites of arachidonic acid based on the specificities of ultraviolet absorption of these various metabolites and is sensitive to the nanogram level. This procedure makes it possible to extract complex mixtures of eicosanoids efficiently with a single step and to analyze them simultaneously by RP-HPLC from biological samples using octadesylsilyl silica extraction column and PGB2 as an internal standard. The cyclooxygenase, products (prostaglandin (PG)D2, PGE1, PGE2, PGF1alpha, PGF2alpha, 6-keto-PGF1alpha, and thromboxane B2 (TXB2)) and lipid peroxidation product, isoprostanes, of arachidonic acid were monitored by one isocratic HPLC system at 195 nm wavelength. The lipoxygenase products (leukotriene(LT)B4, LTC4, LTD4, and 5-hydroxyeicosatetraenoic acid (5-HETE), 12-HETE, 15-HETE) were measured by another isocratic HPLC system at 280 nm for LTs and 235 nm for HETEs. This method provides a simple and reliable way to extract and assess quantitatively the final arachidonic acid metabolites.
12-Hydroxy-5,8,10,14-eicosatetraenoic Acid ; Absorption ; Alprostadil ; Arachidonic Acid* ; Chromatography, High Pressure Liquid* ; Chromatography, Liquid ; Complex Mixtures ; Dinoprost ; Dinoprostone ; Eicosanoids ; Hydroxyeicosatetraenoic Acids ; Isoprostanes ; Leukotriene C4 ; Leukotriene D4 ; Lipid Peroxidation ; Lipoxygenase ; Prostaglandin-Endoperoxide Synthases ; Silicon Dioxide ; Solid Phase Extraction* ; Thromboxane B2

OBJECTIVETo study action of Cryptoporus volvatus ferment substance (CVFS) on leukotriene production of polymorphonuclear leukocytes in rats.

METHODSThe level of slow reaction substance (SRS) and leukotriene B4 (LTB4) in polymorphonuclear leukocytes (PMNs) in rats in vitro were determined with bioassay and HPLC.

RESULTSCVFS 0.9, 2.7 g.kg-1 by ig significantly inhibited SRS and LTB4 production in PMNs in rats in vivo.

CONCLUSIONThe inhibition effect of CVFS on SRS and LTB4 release may be related to its mechanism of anti-inflammation and anti-asthma.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Cell Separation ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Guinea Pigs ; Leukotriene B4 ; metabolism ; Male ; Neutrophils ; metabolism ; Polyporaceae ; chemistry ; Rats ; Rats, Sprague-Dawley ; SRS-A ; metabolism

PURPOSE: Recent studies have shown that the cysteinyl leukotriene (cysLT) of exhaled breath condensate (EBC) could be predictive of inflammatory status and effectiveness of treatment in allergic disease. The aim of this study was to evaluate the inflammation and therapeutic effectiveness of cysLT in EBC in pediatric patients with allergic rhinitis (AR). METHODS: We enrolled 34 healthy children (median age, 4 years 10 months) and 67 AR children (median age, 5 years 1 month). All of the AR patients received intranasal steroid (fluticasone furoate) once daily for 2 weeks. After 2 week of fluticasone furoate treatment, they were classified into 2 groups: the fluticasone furoate (F) and montelukast (M) groups. We treated each group for another 8 weeks. To evaluate the therapeutic effectiveness, we used symptom score (SS) and EBC leukotriene E4 (LTE4). EBC samples were collected with RTube. Each parameter was checked at 0, 2, and 10 weeks of therapy. RESULTS: Most of the AR patients showed clinical improvement with 2- and 10-week fluticasone therapy (F group: 0-week SS, 5.6; 2-week SS, 3.6; 10-week SS, 2.1; P<0.01; M group: 0-week SS, 4.8; 2-week SS, 3.2; 10-week SS, 1.9: P<0.01). LTE4 levels were higher in AR patients than in control subjects (0 week: 87 pg/mL vs. 18 pg/mL) and were reduced after 2 weeks of fluticasone treatment (F group: 90→51.6 pg/mL, P<0.01; M group: 84→46.1 pg/mL, P<0.01). After 10 weeks of treatment, there was no significant difference in the LTE4 level between the F and M groups. CONCLUSION: LTE4 in EBC may be useful for evaluating inflammation and therapeutic effectiveness in patients with allergic rhinitis.
Child* ; Fluticasone ; Humans ; Inflammation* ; Leukotriene E4* ; Rhinitis, Allergic*

OBJECTIVETo observe the effect of montelukast treatment on levels of serum leukotriene B4 and urinary leukotriene E4 in infants with bronchiolitis.

METHODSSeventy-five children who were diagnosed with bronchiolitis between June 2014 and December 2014 were randomly assigned into two groups, one with thirty-eight cases as the montelukast treatment group and another thirty-seven cases as the control group. All of the children were given routine medical treatment. The children in the montelukast treatment group were additionally given montelukast daily (4 mg once a day, for 7 days). The serum leukotriene B4 and urinary leukotriene E4 levels were measured using ELISA before and after treatment. The relationship between serum leukotriene B4 and urinary leukotriene E4 levels was analyzed by Peason correlation analysis.

RESULTSAfter 7 days of treatment, the serum leukotriene B4 and urinary leukotriene E4 levels in the montelukast treatment and control groups were significantly reduced compared with before treatment (P<0.05). The montelukast treatment group showed significantly lower serum leukotriene B4 and urinary leukotriene E4 levels than the control group (P<0.05). The remission time of cough, wheezing and lung wheezes and the length of hospital stay in the montelukast treatment group were significantly shortened compared with the control group (P<0.05). There was a positive correlation between serum leukotriene B4 and urinary leukotriene E4 levels (r=0.723, P<0.05).

CONCLUSIONSMontelukast has a reliable clinical curative efficacy for bronchiolitis in infants, possibly by decreasing serum leukotriene D4 and urinary leukotriene E4 levels.

Acetates ; therapeutic use ; Bronchiolitis ; drug therapy ; metabolism ; Humans ; Infant ; Leukotriene B4 ; blood ; Leukotriene E4 ; urine ; Quinolines ; therapeutic use