Individualized treatment for allergic rhinitis based on key nasal clinical manifestations combined with histamine and leukotriene D4 levels / Tratamento individualizado para a rinite alérgica baseado nas principais manifestações clínicas nasais e dos níveis de histamina e leucotrieno D4

Abstract Introduction The types of allergic rhinitis are roughly classified based on the causative antigens, disease types, predilection time, and symptom severity. Objective To examine the clinical typing and individualized treatment approach for allergic rhinitis and to determine the optimal treatment method for this disease using various drug combination therapies. Methods A total of 108 participants with allergic rhinitis were divided into three groups based on symptoms. Subsequently, each group was further categorized into four subgroups based on the medications received. The efficacy of the treatments was evaluated using the visual analog scale VAS scores of the total and individual nasal symptoms, decline index of the symptom score, histamine and leukotriene levels, and mRNA and protein expression levels of histamine 1 and cysteinyl leukotriene 1 receptors. Results Loratadine + mometasone furoate and loratadine + mometasone furoate + montelukast significantly improved the sneezing symptom and reduced the histamine levels compared with the other combination therapies (p < 0.05). Meanwhile, montelukast + mometasone furoate and montelukast + mometasone furoate + loratadine considerably improved the nasal obstruction symptom and decreased the leukotriene D4 levels compared with the other combination therapies (p < 0.05). Conclusion Clinical symptom evaluation combined with experimental detection of histamine and leukotriene levels can be an objective and accurate method to clinically classify the allergic rhinitis types. Furthermore, individualized treatment based on allergic rhinitis classification can result in a good treatment efficacy.

Resumo Introdução A rinite alérgica é basicamente classificada de acordo com os antígenos causadores, tipos de doença, peridiocidade e gravidade dos sintomas. Objetivo Avaliar os tipos clínicos e a abordagem terapêutica individualizada para cada tipo de rinite alérgica e determinar o método de tratamento ideal utilizando várias terapias de combinação de fármacos. Método Um total de 108 participantes com rinite alérgica foram divididos em três grupos com base nos sintomas. Posteriormente, cada grupo foi subsequentemente categorizado em quatro subgrupos com base nos medicamentos recebidos. A eficácia dos tratamentos foi avaliada utilizando os escores da escala visual analógica EVA dos sintomas nasais totais e individualmente, índice de declínio do escore de sintomas, níveis de histamina e leucotrienos e níveis de expressão de mRNA e proteína dos receptores de histamina 1 e cisteinil-leucotrieno 1. Resultados As associações entre loratadina + furoato de mometasona, assim como a de loratadina + furoato de mometasona + montelucaste melhoraram significativamente o sintoma de espirros e reduziram os níveis de histamina em comparação às outras terapias combinadas (p < 0,05). Por outro lado, a associação montelucaste + furoato de mometasona, assim como a associação montelucaste + furoato de mometasone + loratadina melhoraram consideravelmente o sintoma de obstrução nasal e diminuíram os níveis de leucotrieno D4 em comparação com as outras combinações (p < 0,05). Conclusão A avaliação clínica dos sintomas combinada com a detecção experimental dos níveis de histamina e leucotrieno pode ser um método objetivo e preciso para classificar clinicamente os tipos de rinite alérgica. Além disso, o tratamento individualizado baseado na classificação da rinite alérgica pode resultar no aumento da eficácia do tratamento.

Leukotriene D4 bronchial provocation test for detection of airway hyper-responsiveness in children / 南方医科大学学报

OBJECTIVE@#To explore the value of leukotriene D4 (LTD4) bronchial provocation test (BPT) in detection of airway hyper-responsiveness (AHR) in children.@*METHODS@#A total of 151 children aged 6 to 14 years, including 86 in remission of asthma and 65 with acute bronchitis, who were followed up in our respiratory clinic between November, 2017 and August, 2018. The children were randomly divided into LTD4 group (78 cases) and methacholine (MCH) group (73 cases). In LTD4 group, the 78 children underwent LTD4-BPT, including 46 with asthma and 32 children having re-examination for previous episodes of acute bronchitis; in MCH group, the 73 children underwent MCH-BPT, including 40 with asthma and 33 with acute bronchitis. MCH-BPT was also performed in the asthmatic children in the LTD4 group who had negative responses to LTD4 after an elution period. The major adverse reactions of the children to the two BPT were recorded. The diagnostic values of the two BPT were evaluated using receiver-operating characteristic (ROC) curve.@*RESULTS@#There was no significant difference in the results of basic lung function tests between LTD4 group and MCH group (>0.05). The positive rate of BPT in asthmatic children in the LTD4 group was significantly lower than that in the MCH group (26.1% 72.5%; < 0.05). The positive rate of BPT in children with previous acute bronchitis in the LTD4 group was lower than that in the MCH group (3.1% 15.2%). The positive rate of MCH-BPT in asthmatic children had negative BPT results in LTD4 group was 58.8%, and their asthma was mostly mild. The sensitivity was lower in LTD4 group than in MCH group (0.2609 0.725), but the specificity was slightly higher in LTD4 group (0.9688 vs 0.8485).The area under ROC curvein LTD4 group was lower than that in MCH group (0.635 0.787). In children with asthma in the LTD4 group, the main adverse reactions in BPT included cough (34.8%), shortness of breath (19.6%), chest tightness (15.2%), and wheezing (10.9%). The incidence of these adverse reactions was significantly lower in LTD4 group than in MCH group ( < 0.05). Serious adverse reactions occurred in neither of the two groups.@*CONCLUSIONS@#LTD4-BPT had high safety in clinical application of children and was similar to the specificity of MCH-BPT. However, it had low sensitivity, low diagnostic value, and limited application value in children's AHR detection.

Avaliação do efeito do antagonismo do receptor de leucotrienos na modulação da periodontite experimental em ratos / Cysteinyl leukotriene receptor antagonism modulates experimental periodontitis in rats

Na doença periodontal, quando ativada a resposta imunoinflamatória do hospedeiro, os leucotrienos (LTs) participam do processo de lesão tecidual pela quimiotaxia de leucócitos e ativação osteoclástica. O uso de antagonista do receptor de LTs está relacionado com a expressão de moléculas pró-inflamatórias e osteoclastogênese. No entanto, suas implicações na progressão da DP ainda não foram estudadas. Sendo assim, o trabalho teve como objetivo avaliar o efeito de antagonista do receptor de LTs (Montelucaste - MT) na modulação da periodontite experimental induzida pelo método da ligadura em ratos. Ratos Wistar machos (6-8 semanas, 200-250 gramas) foram divididos (12 animais/ grupo) nos seguintes grupos experimentais: Sham - sem ligadura/ sem tratamento (carboximetilcelulose - CMC 0,5%, via gavagem); Periodontite - com ligadura/ sem tratamento; MT 10 - com ligadura/ com tratamento (Montelucaste 10 mg/kg/dia, via gavagem); MT 30 - com ligadura/ com tratamento (Montelucaste 30 mg/kg/dia, via gavagem). Após período experimental (7, 14 e 21 dias) os animais foram submetidos à eutanásia e as hemimandíbulas retiradas para realização da análise de perda óssea alveolar (POA) macroscópica, análise histológica (H&E - histopatológico; picrosirius - colágeno), ELISA (LTB4), atividade de mieloperoxidase (MPO), avaliação de marcadores do estresse oxidativo (glutationa, expressão de proteínas carboniladas e proteínas modificadas por 4HNE) e expressão gênica de receptor do ativador de fator nuclear kappa B (RANK), ligante de RANK (RANKL), osteoprotegerina (OPG), fator de transcrição relacionado ao runt 2 (RUNX2), Colágeno tipo I, receptor de LT 1 (BLT1), receptor 1 de cisteinil-leucotrieno (CisLTR1), LTA4 hidrolase (LTA4H) e LTC4 sintase (LTC4S) (ANOVA, pós teste Sidak, p<0,05). O grupo Periodontite apresentou maior POA em comparação ao grupo Sham nos três períodos experimentais avaliados (p<0,05). MT passou a ser efetivo na redução da POA a partir do 14º dia (p<0,05). Ao analisar a degradação de colágeno, no 21º dia os grupos Periodontite e MT apresentaram maior perda em comparação ao grupo Sham (p<0,05). Não foram encontradas diferenças estatísticas quanto à expressão de LTB4 pelo teste ELISA (p>0,05), bem como na avaliação de marcadores de estresse oxidativo, independente do teste realizado (p>0,05). Em relação à MPO, o grupo Periodontite apresentou valores estatisticamente maiores em comparação ao grupo Sham nos três períodos experimentais (p<0,05). Nos períodos de 7 e 14 dias (30 mg/kg) o grupo MT promoveu diminuição da infiltração granulocítica em comparação ao grupo Periodontite (p<0,05), sem diferença em relação ao grupo Sham (p>0,05). Observou-se que a expressão de colágeno tipo 1 não apresentou diferença significante entre os grupos (p>0,05), e que o grupo MT 30 apresentou maior expressão de RUNX2 em comparação aos grupos Sham e Periodontite (p<0,05). No grupo MT 30 houve uma redução significante da expressão de LTA4H, BLT1 e LTC4S (p<0,05). A expressão de CysLTR1 não diferiu entre os grupos (p>0,05). Conclui-se que o uso de antagonista de receptor de LTs foi efetivo na redução do infiltrado granulocítico e da POA, com aumento de RUNX2.

Correlations of nasal responses to leukotriene D4 and histamine nasal provocation with quality of life in allergic rhinitis

BACKGROUND: The symptoms of allergic rhinitis (AR) greatly affect the quality of life (QoL) in the patients with AR. The correlations of nasal response to leukotriene D4 (LTD4) and histamine nasal provocation with health related QoL in AR are not clear. OBJECTIVE: To evaluate the correlations of nasal response to LTD4 and histamine nasal challenge with QoL in AR. METHODS: Patients randomly underwent LTD4 and histamine nasal challenge tests, completed the rhinoconjunctivitis quality of life questionnaire (RQoLQ), and rating the symptom severity score (total symptom score 4, TSS4) in the previous week. The correlations between nasal challenge tests induced nasal responses and QoL in RQoLQ were analyzed. RESULTS: A total of 25 eligible AR patients enrolled and finished both LTD4 and histamine nasal challenge and completed the questionnaire of RQoLQ. Histamine nasal challenge induced sneezing, increased nasal resistant were correlated with most of the dimensions (general, practical, nasal, eye problems, and quality of sleep, p < 0.05), while LTD4 nasal challenge induced sneeze, increased nasal resistant only correlated with nasal and ocular problems. On the contrary, the severity of the sneeze assessed by TSS4, was not correlated with QoL, while the severity of rhinorrhea, congestion, and nasal pruritus were correlated with nasal and practical problems, and nasal congestion was also correlated with ocular problems (r = 0.60, p = 0.01). CONCLUSION: LTD4 and histamine nasal challenge induced nasal responses were correlated with different clinical symptoms severity and QoL, which can be used as a good diagnosis and evaluation methods for the management of AR.

Effects of leukotriene D4 on proliferation and migration of lung epithelial A549 cells in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of cysteinyl leukotriene (CysLT) receptor agonist leukotriene D4 (LTD4) on proliferation and migration in lung epithelial A549 cells.</p><p><b>METHODS</b>The expression of CysLT1 receptor and CysLT2 receptor was determined by immunofluoresence staining in A549 cells. A549 cells were treated with LTD4 (0.01-100 nmol/L) for 24-72 h. Cell viability was detected by MTT reduction assay. Cell migration was determined by modified scratch and healing model.</p><p><b>RESULTS</b>In A549 cells, CysLT1 receptor and CysLT2 receptor were mainly expressed in the cytoplasm, membrane and few in the nuclei. The treatment of LTD4 (0.01-100 nmol/L) for 24-72 h caused no effect on cell viability (Ps>0.05); when A549 cells were treated with 100 nmol/L LTD4 for 24, 48 and 72 h the cell viability was (103.00±4.46)%，(107.00±9.45)% and (105.00±9.02)% of control, respectively (Ps>0.05). The migration rate of A549 cells after scratching during the first 24 h was markedly greater than that during the second and third 24 h in the same concentration groups; however, no significant difference in migration rate was noticed when the cells were treated with different concentrations of LTD4 (0.01-100 nmol/L)(Ps>0.05). The migration of A549 cells was 1.15-fold, 1.21-fold and 1.06-fold of that of control when the cells were treated with 100 nmol/L LTD4 for 24, 48 and 72 h, respectively (Ps>0.05).</p><p><b>CONCLUSION</b>The proliferation and migration of A549 cells are not changed when treated with 0.01-100 nmol LTD4 for up to 72h.</p>

Leukotriene D4 activates BV2 microglia in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of CysLT receptor agonist leukotriene D4(LTD4) and antagonists on activation of microglia BV2 cells.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>In 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.</p><p><b>CONCLUSION</b>LTD4 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.</p>

Effects of agonist and antagonist of cysteinyl leukotriene receptors on differentiation of rat glioma C6 cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the role of cysteinyl leukotriene (CysLT) receptors in the differentiation of rat glioma C6 cells.</p><p><b>METHODS</b>Rat glioma C6 cells were treated with the agonist LTD(4), the CysLT(1) receptor antagonist montelukast and the differentiation inducer forskolin. Cell morphology and GFAP protein expression were determined after treatments.</p><p><b>RESULT</b>Forskolin (10 μmol/L) induced morphological changes and GFAP protein expression (cell differentiation) in C6 cells, but LTD(4) (0.1-100 nmol/L) did not induce these changes. Montelukast (1 μmol/L) alone did not affect C6 cell differentiation, while it induced the differentiation when combined with the LTD(4) (100 nmol/L).</p><p><b>CONCLUSION</b>The CysLT(2) receptor may modulate the differentiation of rat glioma C6 cells.</p>

Changes of leukotriene D4 in nasal discharge and plasma of patients with persistent allergic rhinitis and the effects of antihistamine / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the changes of leukotriene D4 (LTD4) in nasal discharge and plasma of patients with persistent allergic rhinitis (AR) and the effects of antihistamine.@*METHOD@#The investigation was a prospective, randomized controlled trial. Forty AR patients (group C) were divided randomly into two subgroup. One group received oral antihistamine 10 mg everyday for one week (group CA) and another group received no loratadine tablets 10 mg everyday for one week (group CB). Fifteen age matched healthy (group D) people were enrolled as control. The level of LTD4 and interleukin-5 (IL-5) in both nasal discharge and plasma by using enzyme linked immunosorbent assay (ELISA) and enzyme immunoassay (EIA), cell counts and cell differentials in nasal discharge, were measured before and after three month. The clinical symptom and life quality scores of group C were also investigated.@*RESULT@#The concentrations of LTD4 in nasal discharge [(794 +/- 305) pg] and plasma [(5219 +/- those in group D [(347 +/- 169) pg, (2283 +/- 489) ng/L, all P 1185) ng/L] in group C were significantly higher than those in group D [(347 +/- 169) pg, (2283 +/- 489) ng/L, all P 0.05).@*CONCLUSION@#The results suggested that LTD4 was involved in airway inflammation in AR. Antihistamine was not effective enough in decreasing the levels of LTD4 in both nasal discharge and plasma of AR patients.

Method for screening cysteinyl leukotriene receptor 2 antagonists and preliminary screening of compounds / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a method for screening cysteinyl leukotriene receptor 2 (CysLT(2)) antagonists and to preliminarily screen a series of synthetic compounds.</p><p><b>METHODS</b>Rat glioma cell line (C6 cells) highly expressing CysLT(2) receptor was used. Intracellular calcium concentration was measured after stimulation with the agonist LTD(4),which was used to screen compounds with antagonist activity for CysLT(2) receptor. Bay u9773, a CysLT1/CysLT(2) receptor non-selective antagonist, and AP-100984, a CysLT(2) receptor antagonist, were used as control.</p><p><b>RESULT</b>PT-PCR showed a higher expression of CysLT(2) receptor in C6 cells. LTD(4) at 1 mumol/L significantly increased intracellular calcium in C6 cells; the maximal effect was about 37.5% of ATP, a positive stimulus.LTD(4)-induced increase of intracellular calcium was blocked by CysLT(2) receptor antagonists, but not by CysLT(1) receptor antagonists. Among the synthetic compounds, D(XW-)1,2,13,23,29 and 30 inhibited LTD(4)-induced increase of intracellular calcium.</p><p><b>CONCLUSION</b>LTD(4)-induced change in intracellular calcium in C6 cells can be used as a screening method for CysLT(2) receptor antagonists. The compounds, D(XW-)1,2,13,23,29 and 30, possess antagonist activity for CysLT(2) receptor.</p>

Effects of cysteinyl receptor agonist and antagonists on rat primary cortical neurons / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine the effect of cysteinyl receptor agonist leukotriene D(4) (LTD(4)) and its antagonists on rat primary neurons.</p><p><b>METHODS</b>In the primarily cultured rat cortical neurons, the neuron injury was evaluated by measuring intracellular calcium, 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyl tetrazolium bromide (MTT) reduction, and propidium iodide (PI) and Hoechst 33258 staining. The in vitro ischemic injury was induced by oxygen-glucose deprivation (OGD) for 1.5 h and reperfusion for 24 h.</p><p><b>RESULT</b>LTD(4) (0.01-1 micromol/L) did not induce the elevation of intracellular calcium, neuron viability changes and neuron death. OGD-induced injury was not significantly ameliorated by the CysLT(1) receptor antagonists, pranlukast (0.2-10 micromol/L) and montelukast (0.2-10 micromol/L), as well as by the CysLT(1)/CysLT(2) receptor non-selective antagonist, BAY u9773 (0.02-1 micromol/L).</p><p><b>CONCLUSION</b>Neither agonist nor antagonists of cysteinyl receptors have the effects on the viability and ischemic-like injury in rat primary neurons.</p>

Cysteinyl leukotriene receptor 1 antagonist pranlukast modulates differentiation of SK-N-SH cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine whether cysteinyl leukotriene receptor agonist LTD(4) and cysteinyl leukotriene receptor 1 (CysLT(1)) antagonist pranlukast affect the differentiation of human neuroblastoma SK-N-SH cells.</p><p><b>METHODS</b>SK-N-SH cell morphological changes induced by LTD(4), pranlukast and LTD(4) + pranlukast were observed with retinoid acid (RA) as the positive control. The expressions of CysLT(1) and CysLT(2) receptors were detected by immunoblotting analysis, and the expression of microtubule-associated protein-2 (MAP-2), a neuron marker, was detected by fluorescent immunostaining.</p><p><b>RESULT</b>The immunoblotting results showed that SK-N-SH cells expressed CysLT(1) receptor moderately, and CysLT(2) receptor highly. The morphological results showed that RA, pranlukast and LTD(4) + pranlukast induced the compaction of the cell bodies and the outgrowth of neurites, while LTD(4) had no significant effect. The immunostaining results showed that MAP-2 was distributed in the cell bodies in control or pranlukast-treated cells; it was distributed in cell bodies and neuritis in RA-treated cells. Pranlukast increased the numbers of MAP-2-positive cells.</p><p><b>CONCLUSION</b>The CysLT(1)receptor antagonist pranlukast modulates the differentiation of SK-N-SH cells.</p>

Influence of Vasoactive Substances on the Regulation of Cardiac ATP-Sensitive Potassium Channel Activity

BACKGROUND AND OBJECTIVES: It has been known that various vasoactive agents are involved in the regulation of cardiac function through the modification of the K+ channel activities, including the ATP-sensitive K+ channel (KATP). We examined the effects of several vasoactive agents on the cardiac KATP currents in isolated cardiac myocytes. MATERIALS AND METHODS: Ventricular myocytes were isolated from the hearts of ICR mice by enzymatic digestion. The channel currents were recorded by the excised inside-out and cell-attached patch clamp configurations. RESULTS: In the excised inside-out patches, bradykinin (BRK; 1-10 micrometer) and prostaglandin I2 (PGI; 10-50 micrometer) did not affect the channel activities, whereas the vasodilators increased the attenuated channel activities in the presence of 100 micrometer ATP. BRK and PGI in parallel shifted the dose-response curves of ATP (1-1,000 micrometer), and this inhibited the KATP currents to the right. Endothelin (ET-1; 0.1-1 nM) and leukotriene D4 (LTD; 3-10 micrometer) decreased the channel activities immediately after making the inside-out patches. However, the vasoconstrictors did not affect the attenuated channel activities by ATP. In the cell-attached patches, both BRK and PGI increased the channel activities and these effects were markedly attenuated by glibenclamide (50 micrometer). ET-1 and LTD did not affect the baseline channel activities in the cell-attached patches, but they markedly attenuated the dinitrophenol-induced activities. CONCLUSION: It was inferred that certain vasoactive substances are involved in the regulation of cardiac KATP channel activities, and that bradykinin and PGI2 enhance the channel activities, and ET-1 and LTD4 inhibit the channel activities.

Effect of shuanglong capsule on content of leukotrienes compound of lung tissues in asthmatic rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>SD 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSION</b>SLC 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.</p>

MUC2/5AC Expression and Mucin Secretion through Leukotriene Receptor in Human Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Mucin gene expression and mucin production are highly increased during inflammatory airway disorders such as, asthma, chronic bronchitis and sinusitis. Cytokines, lipopolysaccharides and other inflammatory mediators are related with secretion and production of mucin. However, among of inflammatory mediators, the relation of leukotrienes and mucin genes expression is not clear. The aim of this study is to evaluate MUC2/5AC genes expression and mucin secretion through leukotriene receptor in human airway epithelial cells. SUBJECTS AND METHOD: The effect of Leukotriene D4 and leukotriene receptor antagonist, pranlukast hydrate (ONO-1078) on the regulation of MUC2/5AC gene expression and mucin secretion was observed in the human airway NCI-H292 epithelial cells. The mRNA levels of MUC2/5AC and the amount of mucin protein were determined by reverse transcription-polymerase chain reaction (RT-PCR) and immunoassay. RESULTS: Leukotriene D4 upregulated MUC2/5AC gene expression and mucin secretion on a dose dependent pattern. Pranlukast hydrate (ONO-1078, 100 micrometer) downregulated the leukotriene D4-mediated MUC2/5AC gene expression and mucin secretion. CONCLUSION: These results suggest that the leukotriene receptor system is one of the expression mechanisms of MUC2/5AC genes and mucin secretion.

Effect of montelukast-loratadine combination versus montelukast alone in asthma [an experimental and clinical study]

Montelukast is a potent, oral, specific leukotriene D 4 receptor antagonist recently approved for the treatment of chronic asthma in patients aged 6 years and older. Loratadine is a long acting selective HI - receptor antagonist devoid of significant sedative or anticholinergic properties. In addition to its activity as an HI - receptor antagonist, loratadine has demonstrated other anti-allergic properties. In this work, the in-vitro experiments were performed to compare the effectiveness of combination of montelukast [1.6 x 10 [-6] M/L] and loratadine [1.6 x 10 [-6]6 M/L] on antigen-induced contractions in sensitized isolated guinea pig tracheal spirals. They revealed that pretreatment with montelukast and loratadine either combined or each alone produced a significant reduction in the ovalbumin-induced contractionsin the doses 0.1, 1, 10 and 100 mg/ml. The EC5O values of ovalbumin-induced contractions were 27, 139.19 and 3.34 X 10 [12] ug/ ml after pretreatment with morclukast. loratadine and their combination, respectively. It has been found that montelukast-loratadine combination was more effective than montelukast alone. Results of the clinical study demonstrated that Montelukast alone and Montelukast-loratadine administered once daily at bedtime to asthmatic patients of mild to moderate asthma and are sub-optimally controlled with glucocorticoids and theophylline showed significant improvements in objective and subjective measurements of asthma control. Montelukast-loratadine demonstrated significant improvements over Montelukast alone in FEV 1 [primary end point], PEFR, daily B2 agonist use, daytime symptom scores [secondary end points] and nocturnal asthma symptoms score. In conclusion, montelukast-loratadine combination provides additional benefit compared with montelukast alone in mild to moderate asthma. In addition, montelukast provides a replacement therapy for glucocorticoids and theophylline in these patients

The Effects of Leukotriene Receptor Antagonist on Nasal Mucous Membrane in Allergic Model of Guinea Pigs / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The leukotriene (LT) receptor antagonist is subject to an on-going study of allergic rhinitis, nasal polyposis and chronic paranasal sinusitis. This study was designed to evaluate the change of nasal patency and morphological changes by assessing the role of 4-oxo-8-benzopyren-hemihydrate (ONO-1078, BH), a cysLT1 receptor antagonist to treatment of allergic rhinitis. MATERIALS AND METHOD: Sixty-five guinea pigs (GPs) were divided into 3 groups: 15 for the control group, 25 for sensitized GPs group and 25 for nonsensitized GPs group. Sensitized GPs were actively sensitized by intraperitoneal injection of 10 mug DNP-As containing 1 mL Al (OH)3 and booster injections were given intraperitoneally 2, 4 and 6 weeks after the initial immunization. Measurements of nasal volume were made by acoustic rhinometry. Also transmission electron microscopy was performed to investigate ultrastructural changes of the nasal mucosal membrane in the LTD4 administrated GPs and the BH treated GPs. RESULTS: Acoustic rhinometry revealed that the changes of nasal volume showed significant reduction at 30 minutes and 6 hours after instillation of LTD4 in nonsensitized guinea pigs (GPs). However, neither nonsensitized nor sensitized GPs with systemic administration of BH showed any changes in nasal patency. Many neutrophils and eosinophils were seen in perivascular space after local administration of LTD4 in control GPs. However there are no eosinophil infiltration into the subepithelial space in BH treated GPs in both nonsensitized and sensitized group. CONCLUSION: The results suggest that BH might be a potent LT receptor antagonist in the allergic model of GPs, which reduces nasal blockage and block chemotaxis of eosinophils to the mucous membrane of the nose.

Inhibition of Cryptoporus volvatus ferment substance on release of leukotriene B4, C4 and D4 from neutrophils in rats in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study inhibitory the effects of Cryptoporus volvatus ferment substance(CVFS) on leukotriene production in vitro from neutrophils in rats.</p><p><b>METHODS</b>Neutrophil 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).</p><p><b>RESULT</b>CVFS 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.</p><p><b>CONCLUSION</b>The results suggest that suppression of leukotriene release may be a mechanism of the anti-inflammation and anti-asthma effects of CVFS.</p>

The Difference between the Concentrations of Arachidonic Acid Metabolites in the Cholesterol Granuloma and Granulation Tissues in Chronic Otitis Media / 대한이비인후과학회지

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.

Effect of Mahuangbujaseshintang and Soshihotang in animal model of allergic rhinitis / 천식및알레르기

BACKGROUND AND OBJECTIVE: Mahuangbujaseshintang (MBST) and soshihotang (SST) have been used for treatment of chronic disease of respiratory tract. It is necessary to clarify the mechanism of anti-allergic effects and to standardize the extracts. MATERIALS AND METHODS: The effects of MBST and SST were evaluated on histamine release in rat mast cells ex vivo. Several hours after administration of the extracts, mast cells were stimulated by DNP-ascaries and histamine contents were measured. Time course structural change of the cells was examined by dynamic study. In order to evaluate the effect of the extracts on the nasal patency, acoustic rhinometry was performed after administering of leukotriene D4 to both nasal cavities of guinea pig (GP). We examined the effects of the extracts with double-blind study, and also studied change of nasal patency after challenge of antigen by acoustic rhinometry in patients with allergic rhinitis. RESULTS: MBST at 4 hr and SST at 3 hr after oral administration remarkably inhibited histamine release from rat mast cells in a dose-dependent manner. MBST-treated GPs failed to show bi-phasic phenomena which indicated to reduce nasal volume at the time of early and late phases in allergic inflammation. Both groups of patients who took MBST and SST for 1 week or 2 weeks showed significant decreased symptom severity index (SSI) from treatment week 2 (p<0.05). The percent volume change after challenge of the antigen was decreased in 31 patients who took the extracts for 2 weeks. CONCLUSION: We can conclude that the herb medicine of MBST and SST may be effective for allergic rhinitis.