Stem cell과 Myeloperoxidase가 스티렌, 하이드로퀴논 및 트리클로로에틸렌에 의한 림프구의 자매염색분체 교환과 소핵체 유도에 미치는 영향

이경재,김형아,신민정,성재혁,박정일,한훈,이세훈 대한산업의학회 2001 대한직업환경의학회지 Vol.13 No.3

목적 : 스티렌, 하이드필퀴논 및 트리클로로에틸렌 (TCE)이 사람의 stem cell과 human myeloperoxidase (MPO)에 의해 대사성 활성화되는 지를 규명하고자 스티렌, 하이드로퀴논 및 TCE에 사람의 stem cell 또는 MPO 효소의 첨가가 이들 화학물질에 의한 자매염색분체교환(SCE)과 소핵체 (MN) 빈도에 미치는 영향을 관찰하였다. 방법 : 건강한 남자의 전혈에서 림프루를 분리하여 72시간동안 이중배양하되 배양개시 24시간만에 0.05mM 하이드로퀴논, 1.50 mM 스티렌, 혹은 1.50mM TCE를 전체용량이 30 ㎕가 되도록 아세톤에 희석하여 배양액에 주입하였고 대조군은 아세톤으로 처리하였다. 화학물질 처리 후 즉시 1.3×106 및 2.6×106 cells/ml 농도의 제대혈액으로부터 나온 stem cell 세포액의 상층액이나 1.0 및 2.0 unit의 human myeloperoxidase를 H2O2와 함께 첨가하였다. SCE분석을 위한 배양액에는 배양종료 2.5시간 전에 colchicine을 가한 후 수확하여 Giemsa염색을 하여 metaphase 세포에서 SCE빈도를 분석하였다. MN분석을 위한 배양액에는 배양개시 44시간만에 cytochalasin-B를 가하였고 acridine orange 염색 후 이핵체에서 MN수를 분석하였다. 결과 : 1. stem cell이나 MPO 자체는 SCE나 MN의 빈도에 영향을 미치지 않았다. 2. stem cell이나 MPO는 스티렌에 의해 유도되는 SCE의 빈도를 용량-반응관계로 유의하게 증가시켰고, MN빈도의 경우 step cell이나 MPO에 의해 증가되는 경향이 있었으나 2.0 unit MPO를 첨가한 경우에만 첨가하지 않은 경우에 비하여 유의하게 증가하였다. 3. 하이드로퀴논은 stem cell이나 MPO가 없는 상태에서도 대조군에 비하여 SCE빈도가 대조군에 비하여 높았다. stem cell이나 MPO는 하이드로퀴논에 의한 SCE 빈도를 용량 반응관계로 유의하게 증가시켰지만, MN의 경우에는 증가시키는 경향만 있을 뿐 유의한 차이는 아니었다. 4. TCE자체는 SCE나 MN빈도를 증가시키지 않았다. stem cell은 1.3 ×106 및 2.6 × 106 cells/ml 농도 모두에서 SCE빈도를 유의하게 증가시켰고 MPO는 2.0 unit농도에서만 유의하게 증가시켰다. stem cell이나 MPO모두 TCE에 의한 MN빈도를 증가시키는 경향이 있었으나 유의한 차이는 아니었다. 결론 : 저자들은 스티렌, 하이드로퀴논, 및 트리클로로에틸렌에 의해 유도되는 자매염색분체교환과 소핵체의 빈도가 사람의 stem cell이나 myeloperoxidase에 의해 증가됨을 발견하였으며, 이러한 결과는 myeloperoxidase가 이들 물질의 대사성활성화에 관여함을 암시하고, 또한 아마도 이 물질들의 골수독성과 관련이 있는 것이라고 제시된다. Objectives : The objective of this study was to identify the possible role of stem cell and myeloperoxidase (MPO) in the metabolic activation of styrene, hydroquinone and trichloroethylene, by investigating the effects of stem cell from umbilical cord blood and MPO on the frequency of sister chromatid exchange (SCE) and micronuclei (MN) induction in cultured human peripheral lymphocytes exposed to hese chemicals. Methods : Islated lymphocytes from whole blood were cultured for 72 hours. The cells were treated with 1.50 mM styrene, 50 μM hydroquinone and 1.50 mM trichloroethylene dissolved with acetone (30㎕ in total volume) at 24 hours after the beginning of culture. Control group was treated with acetone only. Immediately after adding these chemicals, 1.3×106 cells/ml and 2.6×106 cells/ml stem cell/ml stained with Giemsa's solution, and acridine orange for sister chromatid exchange, and for micronucleus analysis, respectively. Results : The results were as follows: 1) Myeloperoxidase and stem cell did not significantly affect the frequencies of SCE or MN in the control group. 2) The frequency of SCE or MN with exposure to styrene did not different from control in the absence of stem cell or MPO. Sister chromatid exchange induced by styrene was significantly increased by adding stem cell or MPO in dose-dependent relationship. The frequency of MN induced by styrene significantly increased in the presence of 2.0 unit MPO. 3) The frequency of SCE was significantly increased with exposure to hydroquinone than acetone treated control in the absence of stem cell or MPO. Sister chromatid exchange induction by hydroquinone significantly increased dose-dependently in the presence of stem cell or MPO. There was a tnendency of increase of the MN frequency induced by hydroquinone in the presence of stem cell or MPO, but not significant. 4) It was found that trichloroethylene itself did not increase SCE or MN frequency. Frequency of SCE induced by trichloroethylene was significantly increased with adding stem cell (low and high) and 2.0 unit MPO. Even though them cell or MPO increased the frequency of MN of lymphocyte exposed to trichloroethylene, the difference was not significant. Conclusions : Authors found that the frequencies of both sister chromatid exchange and micronucleus induced by styrene, hydroquinone, and trichloroethylene were increased significantly with the treatment of stem cell or myeloperoxidase. It was suggested that myeloperoxidase may therefore play an important role in the metabolic activation of styrene, hydroquinone, and trichloroethylene and myeloperoxidase probably be involved in the myelotoxicity of these chemicals.

산란계(Dakalbwarren)의 연령에 따른 myeloperoxidase와 leukocyte peroxidase 활성에 관한 연구

전승기,강창원,이호일,Chon, Seung-ki,Kang, Chang-won,Lee, Ho-il 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.3

This study was undertaken to measure the activity of myeloperoxidase and leukocyte peroxidase of hen. 70 hens were decapitated to observe the activity of enzymes according to ages. The activity of peroxidase by the Lowry's method with bovine serum albumin as standard. Gel filtration chromatography was carried out of sephacryl S-300 column. The results obtained were summarized as follows; 1. The mean of specific activity of myeloperoxidase and leukocyte peroxidase was 16.80(units/mg) and 15(units/mg), respectively. 2. The specific activity of myeloperoxidase in 35 days hen was significantly increased and showed almost the same level of activity to 350 days hen. 3. The specific activity of leukocyte peroxidase in 35 days hen was significantly increased and showed a little increased tendency from 210 days to 350 days hen. 4. On the sephacryl S-300 column chromatography, two separated peaks of myeloperoxidase activity were observed. The molecular weights of myeloperoxidase were 57,000 dalton and 13,700 dalton.

심장수술시 Myeloperoxidase 생성의 증가와 심장 및 폐에 대한 영향

최석철(Seok-Cheol Choi) 대한의생명과학회 2000 Biomedical Science Letters Vol.6 No.4

심장수술에 따른 백혈구의 활성화는 여러 염증성 물질을 생성시켜 수술 후 병태생리에 깊이 관여한다. 본 연구는 심장수술 동안 백혈구의 과립성분인 myeloperoxidase의 농도 변화와 심장 및 폐에 대한 영향을 규명하기 위해 시행되었다. 심장수술 전후로 말초혈액 총 백혈구 수 및 분획세포 수와 관상정맥 혈액의 myeloperoxidase 농도 (MPO) 및 troponin-T 농도 (TnT), 그리고 폐혈관 저항을 측정하였다. 수술 직후, 호중구 수의 상승에 기인한 총 백혈구 수의 증가가 있었고 (p<0.01), MPO 및 TnT 역시 증가하였으나 (p<0.05), 폐혈관 저항은 감소하였다 (p<0.05). 수술 후, MPO는 TnT, 총 백혈구 수, 호중구 수, 그리고 심장수술 소요시간과 각각 양의 상관관계가 있었다 (p<0.05). 수술 후 폐혈관 저항은 총 백혈구 수 및 호중구 수와 각각 음의 상관성이 있었다 (p<0.05). 본 연구의 결과들은 심장수술이 호중구로 부터 MPO 분비를 증가시키며, 유리된 MPO는 폐보다는 심장에 유해한 효과를 미칠 수 있고 호중구의 또 다른 기전이 수술 후 폐 손상에 관여할 수 있음을 시사하고 있다. Leukocyte activation with cardiac surgery procedures produces various inflammatory substances and involves in postoperative pathophysiology. The present study was carried out to elucidate changes in leukocyte myeloperoxidase level and effect on the heart and lung during cardiac operation. Total leukocyte and differential counts in peripheral blood, myeloperoxidase (MPO) and troponin-T concentrations (TnT) in coronary sinus blood, and pulmonary vascular resistance (PVR) were measured at preoperative and postoperative period. The parameters were compared between sampling periods, and relationship was investigated between MPO and each variable. At the end of operation, there were leukocytosis with neutrophilia (p<0.01), and increases of MPO and TnT concentrations (p<0.05), but decrease in PVR (p<0.05). MPO had a positive correlation to TnT, total leukocyte, neutrophil, or operative times (p<0.05), whereas PVR had a negative relationship to total leukocyte or neutrophil counts (p<0.05). These results indicate that cardiac surgery leads to elevated liberations of myeloperoxidase from neutrophils and may harmfully affect myocardium.

흡입용 스테로이드가 기관지천식 환아의 객담내 Interleukin-8과 Myeloperoxidase에 미치는 영향

이강호,이지영,이광민,이영희,이미애,이신재,박강서 대한 소아알레르기 및 호흡기학회 2001 소아알레르기 및 호흡기학회지 Vol.11 No.4

Aged Garlic Supplement Protects Against Lipid Peroxidation in Hypercholesterolemic Individuals

Xing Lin Ho,Shin Yih Tsen,Mei Yen Ng,Wei Ni Lee,Aileen Low,Wai Mun Loke 한국식품영양과학회 2016 Journal of medicinal food Vol.19 No.10

The health benefits of raw garlic intake has been extensively studied, but little is known about the biological effects of aged garlic consumption. A randomized, placebo-controlled, parallel-arm, double-blinded trial involving 41 hypercholesterolemic individuals was conducted to simultaneously examine and compare the blood lipid lowering and antioxidant effects after acute and extended exposures to aged and raw garlic supplements (1080 mg daily). Aged and raw garlic did not affect blood lipid concentrations in these hypercholesterolemic participants after acute and 13-week supplementation. The plasma and urinary F2-isoprostanes concentrations were significantly decreased after 13 weeks of aged garlic treatment. Aged garlic supplementation over 13 weeks also significantly decreased serum lipid hydroperoxide concentration and myeloperoxidase activity. Raw garlic treatments did not affect the F2-isoprostanes concentrations in blood plasma and urine, and lipid hydroperoxides in blood sera. Acute effects on the measured parameters were absent for both garlic treatments. In separate in vitro experiments, aqueous methanolic extract of aged garlic inhibited F2-isoprostanes formation and myeloperoxidase activity in freshly isolated human neutrophils to a greater extent than the raw garlic extract and S-allylcysteine at equivalent dosing concentrations. The aged garlic preparation was found to contain significantly higher total phenolic and S-allylcysteine contents than the raw garlic precursor. Our data showed that supplementation with aged garlic, not its raw garlic precursor, reduced oxidative stress and alleviated lipid peroxidation, possibly via the inhibition of myeloperoxidase. The differential antioxidant actions of the aged and raw garlic may be related to their different total phenolic contents and, to a lesser extent, their S-allylcysteine contents.

Generation of lgG Polymers by the Myeloperoxidase-H₂O₂-Catechol System and Their Stimulatory Effect on Oxygen Radical Production from Human Polymorphonuclear Leukocytes

Kim, Yong Sik,Chung, Myung Hee,Lim, Jung Kyoo Seoul National University 1987 The seoul journal of medicine Vol.28 No.3

Histiocytoid Sweet Syndrome in a Child without Underlying Systemic Disease

( Seung Dohn Yeom ),( Hye Soo Ko ),( Jong Hyuk Moon ),( Min Ji Kang ),( Ji Won Byun ),( Gwang Seong Choi ),( Jeonghyun Shin ) 대한피부과학회 2017 Annals of Dermatology Vol.29 No.5

Sweet syndrome (acute, febrile, neutrophilic dermatosis) is characterized by the acute onset of an eruption of painful nodules or erythematous or violaceous plaques on the limbs, face and neck. These symptoms are accompanied by fever. The diagnostic features include histopathological findings of dermal neutrophilic infiltration without leukocytoclastic vasculitis or peripheral blood leukocytosis. Sweet syndrome is associated with infection, malignancies, autoimmune disease, pregnancy, and drugs. Patients with Sweet syndrome demonstrate a complete and rapid response to systemic steroid administration. Recently, a distinct variant of Sweet syndrome was reported, termed “histiocytoid Sweet syndrome”, in which the infiltration of myeloperoxidase-positive histiocytoid mononuclear cells are observed (in contrast to the infiltration of neutrophils). The other clinical features are similar to those of classic Sweet syndrome. Pediatric Sweet syndrome is uncommon, and the histiocytoid type is even rarer. To date, four cases of histiocytoid Sweet syndrome have been reported in children. Herein, we describe a case of histiocytoid Sweet syndrome in an otherwise healthy 10-yearold boy with no underlying systemic disease in whom nonsteroidal, anti-inflammatory drug treatment was successful. (Ann Dermatol 29(5) 626∼629, 2017)

Interstitial Lung Disease and Diffuse Alveolar Hemorrhage, the Two Key Pulmonary Manifestations in Microscopic Polyangiitis

( Min Jung Kim ),( Kichul Shin ) 대한결핵 및 호흡기학회 2021 Tuberculosis and Respiratory Diseases Vol.84 No.4

Microscopic polyangiitis (MPA) is an antineutrophil cytoplasmic antibody (ANCA)-associated necrotizing vasculitis, which mainly affects small vessels in various organs, especially the lungs. The two key pulmonary manifestations, interstitial lung disease (ILD) and diffuse alveolar hemorrhage (DAH), increase the morbidity and death rate of patients with MPA. ILD is more common in MPA than in other ANCA-associated vasculitis subsets and is primarily associated with myeloperoxidase-ANCA. Unlike alveolar hemorrhage due to pulmonary capillaritis, ILD can initially manifest as isolated pulmonary fibrosis. Of note, its most frequent radiographic pattern is the usual interstitial pneumonia pattern, similar to the characteristic pattern seen in idiopathic pulmonary fibrosis. In this review we present the pathogenesis, clinical manifestations, and radiographic and histopathologic features of ILD and DAH in MPA. We also briefly summarize the outcome and therapeutic options for the two conditions.

심장수술시 Myeloperoxidase 생성의 증가와 심장 및 폐에 대한 영향

최석철 대한의생명과학회 2000 Journal of biomedical laboratory sciences Vol.6 No.4

심장수술에 따른 백혈구의 활성화는 여러 염증성 물질을 생성시켜 수술 후 병태생리에 깊이 관여한다. 본 연구는 심장수술 동안 백혈구의 과립성분인 myeloperoxldase의 농도 변화와 심장 및 폐에 대한 영향을 규명하기 위해 시행되었다. 심장수술 전후로 말초혈액 퐁 백혈구 수 및 분획세포수와 관상정맥 혈액의 myeloperoxidase 농도 (MPO) 및 troponin-T 농도 (TnT), 그리고 폐혈관 저항을 측정하였다. 수술 직후, 호중구 수의 상승에 기인한 총 백혈구 수의 증가가 있었고 (p<0.01), MPO 및 TnT 역시 증가하였으나 (p<0.05), 폐혈관 저항은 감소하였다 (p<0.05). 수술 후, MPO는 TnT, 총 백혈구 수, 호중구 수, 그리고 심장수술 소요시간과 각각 양의 상관관계가 있었다 (p<0.05), 수술 후 폐혈관 저항은 총 백혈구 수 및 호중구 수와 각각 음의 상관성이 있었다 (p<0.05). 본 연구의 결과들은 심장수술이 호중구로 부터 MPO 분비를 증가시키며, 유리된 MPO는 폐보다는 심장에 유해한 효과를 미칠 수 있고 호중구의 또 다른 기전이 수술 후 폐 손상에 관여할 수 있음을 시사하고 있다. Leukocyte activation with cardiac surgery procedures produces various inflammatory substances and involves in postoperative pathophysiology. The present study was carried out to elucidate changes in leukocyte myeloperoxidase level and effect on the heart and lung during cardiac operation. Total leukocyte and differential counts in peripheral blood, myeloperoxidase (MPO) and troponin-Tconcentrations (TnT) in coronary sinus blood, and pulmonary vascular resistance (PVR) were measured at preoperative and postoperative period. The parameters were compared between sampling periods, and relationship was investigated between MPO and each variable. At the end of operation there were leukocytosis with neutrophilia (p<0.01), and increases of MPO and TnT concentrations (p<0.05), but decrease in PVR (p<0.05). MPO had a positive correlation to TnT, total leukocyte, neutrophil, or operative times (p<0.05), whereas PVR had a negative relationship to total leukocyte or neutrophil counts (p<0.05). These results indicate that cardiac surgery leads to elevated liberations of myeloperoxidase from neutrophils and may hannfully affect myocardium.

저산소성 허혈성 뇌손상을 유발시킨 어린 백서에서 Myeloperoxidase 측정 검사

양혜정(Hae Joung Yang),피대훈(Dae Hun Pee),임지혜(Ji Hye Lim),최병민(Byung Min Choi),이기형(Kee Hyoung Lee),은백린(Baik-Lin Eun) 대한소아신경학회 2002 대한소아신경학회지 Vol.10 No.2

목 적 : 예전에는 뇌경색 부위에서 발견되는 백혈구를 단지 뇌손상에 뒤따르는 생리적인 반응으로만 생각하였으나 최근 10년 사이 재관류 손상에서도 백혈구에 의한 염증 작용이 중요한 역할을 한다고 알려지면서 이들의 역할에 대한 연구가 새롭게 이루어지고 있다. PML의 존재는 세포질내 과립에서 분비되는 MPO를 생화학적으로 측정함으로서 증명할 수 있는데 저자들은 미성숙 뇌에서 저산소성 허혈증에 의한 뇌손상이 발생할 때 백혈구가 침윤되는 과정을 MPO 측정 검사를 통하여 관찰하였고, P와 L-selectin 억제제인 Fucoidin을 투여한 후 MPO의 변화를 확인하였다. 방 법 : 생후 7일된 어린 흰쥐를 사용하여 우측총 경동맥을 전기 응고시켜 자르고 8% 산소에 노출시켰다. 저산소 노출 후 회복 시간에 따라 동물을 희생시키고 뇌를 추출하여 뇌조직 1 g당 10mL의 20 mM potassium Phosphate buffer(pH 7.4)를 첨가하여 50초간 분쇄하였다. 각각의 균질회된 조직 샘플들은 4℃에서 20분 동안 원심 분리하여 상층액을 제거한 후, 침전물은 처음 조직량에 따라 조직 1 g당 10 mL의 0.5% cetylditrimethylethyl ammonium bromide(wt/vol)가 첨가된 50 mM potassium phosphate buffer(pH 6.0)를 첨가하였다. Vortex를 이용하여 buffer와 침전물을 잘 섞고 60℃수조에서 120분 동안 방치하였다. 그 후 각 조직 샘플들을 4 watts에서 음파처리하고 4℃에서 15분 동안 원심 분리를 시행하여 상층액을 새로운 tube에 옮겼다. MPO 측정검사를 위한 MPO reaction buffer는 50 mM potassium phosphate buffer(pH 6.0) 100 mL에 0.53 mM odianisidine edihydrochlorde와 0.0005% HO를 섞어 만들었다. MPO reaction solution 2.9 mL에 각각의 MPO sample 0.1 mL을 첨가하여 460 nm의 파장에서 5분 동안 MPO에 다른 흡광도 차이를 관찰하였다. 결 과 : 조직내 백혈구 침윤의 지표로 측정한 MPO 활성도는 대조군에서는 미미하였으나 총 경동백을 절단한 우측 대뇌군에서는 허혈 및 저산소증 유발 후 8시간째부터 현저히 증가되기 시작하여 24시간 경과 후 발현이 가장 높았으며, Fucoidin 50 mg/kg으로 전처치한 약물 실험군에서는 MPO 활성도가 현저히 감소하였다. 결 론 : 백혈구는 미성숙 뇌의 저산소성 허혈증에 의한 뇌손상에서 중요한 매개체 역할을 하며, 백혈구의 생화학적 활성도를 나타내는 MPO 활성도는 저산소성 허혈성 뇌손상의 정도를 측정하는 지표로 이용될 수 있다고 생각된다. Purpose : Neutrophils found around an infarcted area in the brain was once considered as only the physiologic response following the brain injury, but recent studies have shown that inflammatory responses by neutrophils play an important role in the reperfusion injury. The presence of ploymorphonuclear leukocytes(PML) is proven by biochemical assay of myeloperoxidase(MPO) Secreted in the cytoplasmic granules. We observed the process of PML infiltration on hypoxic-ischemic brain injury of immature rats by the assay of MPO activity and changes of the MPO activity after the administration of fucoidin, inhibitor of P- and L-selectin. Methods : We used a well characterized model of the brains of 7 day-old-rats, which had unilateral hypoxic and ischemic injuries(HI). Those injuries were induced by unilateral carotid artery ligation followed by timed exposure to hypoxic inspiratory gas mixture(8% O). MPO activity was measured in the brain tissue homogenates of HI rats(n=18) at 0, 2, 8, 24 and 48 hrs and in rats that received fucoidin immediately before and again after hypoxia(50 mg/kg, n=6) at 8 and 24 hrs. Controls(n=2) were rats with neither hypoxia nor ischemia. The brain samples were homogenized in 20 mM potassium phosphate buffer(pH 7.4) for 50 secs. The homogenate was centrifuged at 14,000 g at 4℃ for 15 mins and the supernatant was discarded. The tissue was pulverized, weighed, and suspended in 1 mL of 50 mM potassium phosphate buffer solution(pH 6.0) contatining 0.5% cetylditrimethylammonium bromide(wt/vol). The tissue was sonicated and centrifuged at 10,000 g for 15 mins. 200 µL of the supernatant was mixed with 1 mL of 50 mM potassium phosphate buffer solution(pH 6.0) containing 10 μL of 1.325 mM o-dianisidine hydrochloride and 170 µL of 3% hydrogen peroxide(vol/vol). Changes in absorbance at 460 nm were measured for 5 mins by using microplate reader. One unit of MPO activity was defined as that degrading 1 µmol peroxide/min at 25℃, and the result was expressed as units of MPO/100 gm tissue. Results : In HI rats, MPO activity increased at 2 hrs after HI and peaked at 24 hrs in the right hemisphere. In rats with fucoidin treatment immediately before and again after hypoxia, the MPO activity significantly decreased in both hemispheres compared with HI rats(P<0.05). MPO activity in the tissue of control rats was insignificant. Conclusion : The dynamic changes of the MPO activity suggest the important role of PMN on hypoxic-ischemic brain injuries in immature rats. MPO activity could be used as an index of the severity of injuries of hypoxic-ischemic brains.