Leukotrienes C4 synthase와 cysteinyl leukotriene receptor 1 유전자 다형성과 한국 소아 천식 표현형 및 임상 지표와의 연관성 연구

심정연,김병주,송영화,강미진,이소연,김효빈,유진호,홍수종 대한소아청소년과학회 2009 Clinical and Experimental Pediatrics (CEP) Vol.52 No.6

Purpose : Cysteinyl leukotrienes are important proinflammatory mediators in asthma. Recently, it was suggested that a promoter polymorphism in the genes encoding for leukotriene C4 synthase (LTC4S), a key enzyme in the leukotriene synthetic pathway, and cysteinyl leukotriene receptor 1 (CysLTR1) might be associated with aspirin-intolerant asthma. We investigated whether polymorphisms in LTC4S and CysLTR1 genes or their interactions were associated with the asthma phenotype, lung function, or bronchial hyperreactivity (BHR) in Korean children. Methods : A total of 856 asthmatic children and 254 non-asthmatic controls were enrolled; a skin prick test, lung function test and bronchial provocation test were performed. Of those enrolled, 395 children underwent exercise challenge tests. The LTC4S A(-444)C and CysLTR1 T(+927)C were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis. Results : Of those enrolled, 699 children were classified as having atopic asthma and 277 children, as having exercise-induced asthma (EIA). LTC4S and CysLTR1 polymorphisms were not associated with atopic asthma, EIA, or asthma per se. Lung function and BHR were not significantly different between the wild type (AA or TT) and the variant (AC+CC or TC+CC) genotypes in asthmatics, atopic asthmatics, and EIA (+) asthmatics, while total eosinophil counts were higher in the variant type of LTC4S than in the wild type in atopic asthmatics. There were no associations between the gene-gene interactions of LTC4S and CysLTR1 genotypes and the asthma phenotypes. Conclusion : LTC4S A(-444)C and CysLTR1 T(+927)C polymorphisms and their gene-gene interactions are not associated with asthma phenotype, lung function, or BHR in Korean children. 목 적 : 류코트리엔은 천식의 병태생리에 중요한 향염증성 매개체이며, 천식이나 운동유발성 천식에서 증가되어 있다. Leukotriene C4 synthase (LTC4S)의 A(-444)C 유전자 다형성과 cysteinyl leukotriene receptor 1 (CysLTR1) T(+927)C 유전자 다형성이 한국 소아의 천식, 아토피 천식 및 운동유발성 천식과 연관이 있는지 알아보고, 폐기능, 기관지과민성, 총IgE 치에 영향을 미치는지 알아보고자 하였다. 방 법 : 총 856명의 천식 환자와 254명의 천식이 없는 정상아를 대상으로 하여 피부반응검사, 폐기능, 메타콜린 기관지 유발검사, 총 IgE 검사를 실시하였고, 천식 환자를 아토피 천식(699명), 운동유발성 천식(277명)으로 나누어 유전자 다형성과의 연관성을 조사하였다. LTC4S A9-444)C, CysLTR1 T(+927)C 유전자형은 PCR-restriction fragment length polymorphism 방법으로 분석하였다. 결 과 : LTC4S A(-444)C 및 CysLTR1 T(+927)C 유전자 다형성은 천식, 아토피 천식, 운동유발성 천식과의 연관성이 없었고, 폐기능, PC20, 총IgE에도 차이를 보이지 않았다. 아토피 천식에서 총 호산구수는 변종형 LTC4S 유전자형에서 야생형 보다 높았다. LTC4S A(-444)C와 CysLTR1 T(+927)C의 유전자-유전자 상호 작용도 천식, 아토피 천식, 운동유발성 천식과의 연관이 없었다. 결 론 : 한국 소아 천식의 표현형, 폐기능, 기관지과민성, 총IgE 농도는 LTC4S A(-444)C와 CysLTR1 T(+927)C 유전자 다형성, 혹은 유전자-유전자 상호작용과 연관이 없는 것으로 생각된다.

소아 천식환자에서 Leukotriene C₄ Synthase 유전자 다형태와 Montelukast의 임상적 효과와의 연관성

신경수,김연우,Shin, Kyung Sue,Kim, Youn Woo 대한소아청소년과학회 2005 Clinical and Experimental Pediatrics (CEP) Vol.48 No.7

목 적 : 류코트리엔 수용체 길항제는 천식의 병리 반응에 관여하는 cysteiny leukotriene의 생성과 작용을 억제하여 급성기 천식 증상의 치료와 천식 증상의 조절 요법에 사용할 수 있다. 본 연구에서는 소아 천식환자에서 cysteinyl leukotriene 생성에 관여하는 $LTC_4S$ 유전자 다형태와 류코트리엔 수용체 길항제인 montelukast의 임상적 효과를 조사하여 약물유전학적 연관성 유무를 알고자 하였다. 방 법 : 환자군은 경증 지속성 천식과 중등증 지속성 천식환자 161명을 대상으로 하였고, montelukast 5 mg을 하루에 한 번씩 총 8주 동안 투여하였다. $LTC_4S$ 유전자 다형태는 restriction fragment length polymorphism을 이용하여 조사하였다. 결 과 : 대조군에서 LTC4S 유전자형의 분포는 A/A, A/C, C/C가 각각 74.0%, 22.6%, 3.4%였고, 환자군에서는 A/A, A/C, C/C가 각각 70.8%, 23.6%, 5.6%였다. 두 군의 유전자형 분포는 통계적으로 유의한 차이를 보이지 않았고, $LTC_4S$ 유전자형 분포와 천식의 중증도 사이에도 유의한 차이가 없었다. 반응군에서는 경증 지속성 천식환자가 반응이 없는 군에서는 중등증 지속성 천식환자가 더 많았다. 전체 소아 천식환자군에서는 montelukast에 대한 반응군과 반응이 없는 군 사이에 $LTC_4S$ 유전자형에 따른 차이는 없었다. 경증 지속성 천식환자의 반응군에서 adenine 대립유전자를 가진 환자가 많았으나, 중등증 지속성 천식환자에서는 유전자형에 따른 반응군과 반응이 없는 군의 유의한 차이가 없었다. 결 론 : 본 연구에서 소아 경증 지속성 천식환자의 경우에는 통계적으로 adenine 대립유전자가 montelukast에 대한 임상적 효과를 예측할 수 있는 인자라고 할 수 있으나 전체 소아 천식환자에서는 $LTC_4S$ 유전자 다형태와 montelukast의 임상적 효과와의 연관성은 통계적으로 없었다. Purpose : Cysteinyl leukotrienes are important inflammatory mediators in the pathogenesis of asthma; therefore interruption of cysteinyl leukotrienes by leukotriene receptor antagonists improves clinical symptoms in the management of patients with mild to moderate asthma. We evaluated whether clinical response to montelukast, a leukotriene receptor antagonist, in childhood asthma was predicted by genotypes of leukotriene $C_4$ synthase($LTC_4S$) promoter gene polymorphism. Methods : An 8-week prospective, open trial of montelukast was carried out in 161 children with mild to moderate asthma. Genotyping of $LTC_4S$ gene polymorphism was determined by restriction fragment length polymorphism. Results : The distribution of the $LTC_4S$ genotypes AA, AC, and CC was 70.8 percent, 23.6 percent, and 5.6 percent, respectively in asthma group and 74.0 percent, 22.6 percent, and 3.4 percent, respectively in control group. A statistically significant difference in the distribution of $LTC_4S$ genotype was not observed between the asthma and the control groups, and there was no significant difference between the $LTC_4S$ genotype and asthma severity. The responders to montelukast were significantly prevalent in the mild asthma group(P<0.05). There was no significant difference in the distribution of the responders compared to non-responders within genotype in the total asthma group or the moderate asthma group. However, the responsiveness for montelukast was significant difference within genotype for both AA and AC/CC in the mild asthma group : The AA genotype was more included in the responder group(P<0.05). Conclusion : In the mild persistent asthma group, the A allele of $LTC_4S$ polymorphism may be regarded as a predictable factor for clinical response to montelukast. However, LTC4S polymorphism was not significantly associated with the clinical response to montelukast in asthmatic children.

백서의 격리 폐에서 ATP와 LTC4가 저산소성 폐혈관 수축에 미치는 영향

노지윤 ( Ji Yoon Rho ),신화용 ( Hwa Yong Shin ),김현창 ( Hyun Chang Kim ),이지원 ( Ji Won Lee ),김성덕 ( Seong Deok Kim ) 대한마취과학회 2009 Korean Journal of Anesthesiology Vol.57 No.4

Background: Hypoxic pulmonary vasoconstriction (HPV) is unique to pulmonary circulation but the mechanism remains elusive. Red blood cells (RBCs) are known to augment HPV and to release more ATP as oxygen content falls. Leukotrienes constrict smooth muscle and could be important for the regulation of the pulmonary circulation. Hence we hypothesized that ATP and leukotrienes are mediators of HPV produced during acute alveolar hypoxia. Methods: In forty Sprague-Dawley rats, lungs were isolated and perfused. We administered ATP (10 μM) to the ATP group (n=8), the ATP antagonist, suramin (100 μM) to the suramin group (n=8), leukotriene C4 (LTC4, 5 μg) to the LTC4 group (n=8), the LTC4 antagonist, LY171883 (20 μM) to the LY171883 group (n=8), and LTC4 (5 μg)+ATP (10 μM) to the LTC4+ATP group (n=8) during normoxic ventilation. HPV responses were induced by three hypoxic challenges for 5 minutes separated by 5 minutes of ventilation with a normoxic gas mixture. Baseline pulmonary artery pressure change after exposure to each drug and hypoxic pressor response between a period 21% normoxic gas ventilation and that of 3% hypoxic gas ventilation were measured. Results: ATP and LTC4+ATP increased baseline pulmonary artery pressures but LTC4 did not alter it. ATP did not affect hypoxic pressor response. Suramin, LY171883 and LTC4+ATP inhibited the pressor response to hypoxia. LTC4 increased hypoxic pressor response. Conclusions: In isolated rat lungs, HPV may be mediated by ATP and LTC4 appears more likely to be a modulator than a mediator of HPV. (Korean J Anesthesiol 2009;57:472∼82)

Life Science : Pinusolide Isolated from Biotu orientalis Inhibits 5-Lipoxygenase Dependent Leukotriene C4 Generation by Blocking c-Jun N-Terminal Kinase Pathway in Mast Cells

( Ye Jin ),( Hyun Ok Yang ),( Jong Keun Son ),( Hyeun Wook Chang ) 영남대학교 약품개발연구소 2012 영남대학교 약품개발연구소 연구업적집 Vol.22 No.0

Pinusolide, an herbal medicine isolated from Biota orientalis L. (B. orientalis), inhibited 5-lipoxygenase(5-LO)-dependent leukotriene C4 (LTC4) generation in immunoglobulin E (IgE)/Ag-induced bone marrow-derived mast cells (BMMCs) in a concentration-dependent manner. To clarify the action mechanism of pinusolide on the inhibition of LTC4 generation, we examined the effect of pinusolide on phosphorylation of cytosolic phospholipase A2 (cPLA2), as well as translocation phospho-cPLA2 and 5-LO to nucleus. Inhibition of LTC4 generation by pinusolide was accompanied by a decrease in cPLA2 phosphorylation which occurred via a decrease in intracellular Ca2+ influx and blocking the c-Jun N-terminal kinase (JNK) pathways. However, pinusolide had no effect on extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinas phosphorylation. Taken together, the present results suggest that potent inhibition of 5-LO dependent LTC4 generation bypinusolide requires both suppression of calcium influx and JNK phosphorylation.

Life Science : Citreorosein inhibits degranulation and leukotriene C4 generation through suppression of Syk path way in mast cells

( Yue Lu ),( Ying Li ),( Yurn Dong Jahng ),( Jong Keun Son ),( Hyeun Wook Chang ) 영남대학교 약품개발연구소 2012 영남대학교 약품개발연구소 연구업적집 Vol.22 No.0

The aim of this study was to evaluate whether citreorosein (CIT), a naturally occurring anthraquinone isolated from Polygoni cuspidati (P. cuspidati) radix, modulates degranulation and 5-lipoxygenase (5-LO)-dependent leukotriene C(4) (LTC(4)) generation in mast cells. Cit suppresses both degranulationand the generation of LTC(4) in a dose-dependent manner in stem cell factor (SCF)-mediated mouse bone marrow-derived mast cells (BMMCs). With regard to its molecular mechanism of action, we investigated the effects of CIT on intracellular signaling and mast cell activation employing BMMCs. Binding of SCF to c-Kit on mast cell membranes induced increases in intrinsic tyrosine kinase Syk activity and activation of multiple downstream events including phosphorylation of phospholipase Cγ (PLCγ), mobilization of intracellular Ca(2+), phosphatidylinositol 3-kinase (PI3K), Akt, MAP kinases (MAPKs), translocation of phospho-phospholipase A(2) (PLA(2)) and 5-LO. The results from the biochemical analysis demonstrate that CIT attenuates degranulation and LTC(4) generation through the suppression of multiple step signaling and would be beneficial for the prevention of allergic inflammation.

Imperatorin Suppresses Degranulation and Eicosanoid Generation in Activated Bone Marrow-Derived Mast Cells

정규태,이어진,박나영,김선건,박효현,이지연,이윤주,이은경 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.5

Imperatorin has been known to exert many biological functions including anti-inflammatory activity. In this study, we investigated the inhibitory effects of imperatorin on the production of inflammatory mediators in mouse bone marrow-derived mast cells (BMMC). Imperatorin inhibited degranulation and the generation of eicosanoids (leukotriene C4 (LTC4) and prostaglandin D2 (PGD2)) in IgE/antigen (Ag)-stimulated BMMC. To elucidate the molecular mechanism involved in this process, we investigated the effect of imperatorin on intracellular signaling in BMMC. Biochemical analyses of the IgE/Ag-mediated signaling pathway demonstrated that imperatorin dramatically attenuated degranulation and the production of 5-lipoxygenase-dependent LTC4 and cyclooxygenase-2-dependent PGD2 through the inhibition of intracellular calcium influx/phospholipase Cγ1, cytosolic phospholipase A2/mitogen-activated protein kinases and/or nuclear factor-kB pathways in BMMC. These results suggest that the effects of imperatorin on inhibition of degranulation and eicosanoid generation through the suppression of multiple steps of IgE/Ag-mediated signaling pathways would be beneficial for the prevention of allergic inflammation.

Imperatorin Suppresses Degranulation and Eicosanoid Generation in Activated Bone Marrow-Derived Mast Cells

( Kyu-tae Jeong ),( Eujin Lee ),( Na-young Park ),( Sun-gun Kim ),( Hyo-hyun Park ),( Jiean Lee ),( Youn Ju Lee ),( Eunkyung Lee ) 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.5

Imperatorin has been known to exert many biological functions including anti-inflammatory activity. In this study, we investigated the inhibitory effects of imperatorin on the production of inflammatory mediators in mouse bone marrow-derived mast cells (BMMC). Imperatorin inhibited degranulation and the generation of eicosanoids (leukotriene C4 (LTC4) and prostaglandin D2 (PGD2)) in IgE/antigen (Ag)-stimulated BMMC. To elucidate the molecular mechanism involved in this process, we investigated the effect of imperatorin on intracellular signaling in BMMC. Biochemical analyses of the IgE/Ag-mediated signaling pathway demonstrated that imperatorin dramatically attenuated degranulation and the production of 5-lipoxygenase-dependent LTC4 and cyclooxygenase-2-dependent PGD2 through the inhibition of intracellular calcium influx/phospholipase Cγ1, cytosolic phospholipase A2/mitogen-activated protein kinases and/or nuclear factor-κB pathways in BMMC. These results suggest that the effects of imperatorin on inhibition of degranulation and eicosanoid generation through the suppression of multiple steps of IgE/Ag mediated signaling pathways would be beneficial for the prevention of allergic inflammation.

Saucerneol F, a New Lignan Isolated from Saururus chinensis, Attenuates Degranulation via Phospholipase Cg1 Inhibition and Eicosanoid Generation by Suppressing MAP Kinases in Mast Cells

( Yue Lu ),( Jong Keun Son ),( Hyeun Wook Chang ) 한국응용약물학회 2012 Biomolecules & Therapeutics(구 응용약물학회지) Vol.20 No.6

During our on-going studies to identify bioactive compounds in medicinal herbs, we found that saucerneol F (SF), a naturally oc-curring sesquilignan isolated from Saururus chinensis (S. chinensis), showed in vitro anti-inflammatory activity. In this study, we examined the effects of SF on the generation of 5-lipoxygenase (5-LO) dependent leukotriene C4 (LTC4), cyclooxygenase-2 (COX-2) dependent prostaglandin D2 (PGD2), and on phospholipase Cg1 (PLCg1)-mediated degranulation in SCF-induced mouse bone marrow-derived mast cells (BMMCs). SF inhibited eicosanoid (PGD2 and LTC4) generation and degranulation dose-dependently. To identify the molecular mechanisms underlying the inhibition of eicosanoid generation and degranulation by SF, we examined the effects of SF on the phosphorylation of PLCg1, intracellular Ca2+ influx, the translocation of cytosolic phospholipase A2 (cPLA2) and 5-LO, and on the phosphorylation of MAP kinases (MAPKs). SF was found to reduce intracellular Ca2+ influx by inhibiting PLCg1 phosphorylation and suppressing the nuclear translocations of cPLA2 and 5-LO via the phosphorylations of MAPKs, in-cluding extracellular signal-regulated protein kinase-1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38. Taken together, these results suggest that SF may be useful for regulating mast cell-mediated inflammatory responses by inhibiting degranulation and eicosanoid generation.

Anti-allergic effect of a chloroform-soluble extract of <i>Cinnamomum cambodianum</i> in bone marrow-derived mast cells

Chae, Hee-Sung,Khiev, Piseth,Lee, Hyeong-Kyu,Oh, Sei-Ryang,Chin, Young-Won Informa Healthcare 2012 IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY Vol.34 No.4

<P><I>Cinnamomum cambodianum</I> has been used as a traditional medicine in Cambodia. Its effect on the bone marrow-derived mast cells (BMMCs) mediated allergic response remains unknown. In this study, a chloroform-soluble extract of <I>C. cambodianum</I> was evaluated for its effect on allergic mediators, including prostaglandin D<SUB>2</SUB> (PGD<SUB>2</SUB>), leukotriene C<SUB>4</SUB> (LTC<SUB>4</SUB>), β-hexosaminidase and cyclooxygenase-2 (COX-2) protein, in phorbol 12-myristate 13-acetate (PMA) plus calcimycin-stimulated BMMCs. The results revealed that the chloroform-soluble extract inhibited the production of interleukin-6, PGD<SUB>2</SUB> and LTC<SUB>4</SUB>, and the expression of COX-2 in PMA plus calcimycin-stimulated BMMCs, implying a potential benefit of <I>C. cambodianum</I> in the treatment of allergy.</P>

Original Artide : Saucerneol F, a New Lignan Isolated from Saururus chinensis, Attenuates Degranulation via Phospholipase Cγ1 Inhibition and Eicosanoid Generation by Suppressing MAP Kinases in Mast cells

( Yue Lu ),( Jong Keun Son ),( Hyeun Wook Chang ) 영남대학교 약품개발연구소 2013 영남대학교 약품개발연구소 연구업적집 Vol.23 No.0

During our on-going studies to identify bioactive compounds in medicinal herbs, we found that saucerneol F (SF), a naturally occurring sesquilignan isolated from Saururus chinensis (S. chinensis), showed in vitro anti-inflammatory activity. In this study, we examined the effects of SF on the generation of 5-lipoxygenase (5-LO) dependent leukotriene C4 (LTC4), cyclooxygenase-2 (COX-2) dependent prostaglandin D2 (PGD2), and on phospholipase Cγ1 (PLCγ1)-mediated degranulation in SCF-induced mouse bone marrow-derived mast cells (BMMCs). SF inhibited eicosanoid (PGD2 and LTC4) generation and degranulation dose-dependently. To identify the molecular mechanisms underlying the inhibition of eicosanoid generation and degranulation by SF, we examined the effects of SF on the phosphorylation of PLCγ1, intracellular Ca(2+) influx, the translocation of cytosolic phospholipase A2 (cPLA2) and 5-LO, and on the phosphorylation of MAP kinases (MAPKs). SF was found to reduce intracellular Ca(2+) influx by inhibiting PLCγ1 phosphorylation and suppressing the nuclear translocations of cPLA2 and 5-LO via the phosphorylations of MAPKs, including extracellular signal-regulated protein kinase-1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38. Taken together, these results suggest that SF may be useful for regulating mast cell-mediated inflammatory responses by inhibiting degranulation and eicosanoid generation.