RAW 264.7 세포에 대한 중국산 천연 광물성 섬유 TAFMAG의 독성효과

임영,한진구,김지홍,김현욱,김은경,김경아,장황신 大韓産業醫學會 1999 대한직업환경의학회지 Vol.11 No.3

Objectives : This study was designed to evaluate cytotoxicity of TAFMAG, which is a trade name of natural mineral fiber mined and produced in China. Methods : The cytotoxicity of TAFMAG was evaluated by measuring iron content, lipid peroxidation, erythrocyte hemolysis, and cytotoxicity in vitro. These results were compared with the data of chrystotile and wollastonite as a positive and negative control, respectively. Results : There was significant increase of Fenton activity in TAFMAG and chrysotile with dose-response pattern. The iron chelating agent, desferrioxamine, significantly decreased Fenton activity of the particulates except wollastonite. TAFMAG and chrysotile fibers significantly increased malondialdehyde concentration from lipid peroxidation of the red blood cell membrane. In erythrocyte hemolysis test, TAFMAG & chrysotile had stronger effect on erythrocyte hemolysis than wollastonite with the concentration of 1,000g/ml. Furthermore, TAFMAG was more hemolytic than chrysotile with the concentration of 5,000g/ml. There was a significant cytotoxic effect in TAFMAG and chrysotile on RAW cell compared with wollastonite. Conclusions : In vitro study suggested that TAFMAG may have a similar health hazard as usual asbestos.

시험관 및 동물실험 모델에서 유리규산에 의한 아포프토시스

김지홍,장황신,박영만,안병용,김경아,임영 大韓産業醫學會 1999 대한직업환경의학회지 Vol.11 No.2

Silica exposure results in acute inflammatory response followed by chronic fibrotic change. The mechanism for the maintenance of silica-induced inflammation has not been understood yet. Apoptosis is a morphologically distinct form of programed cell death that plays major role during homeostasis and in many diseases including cancer, acquired immune deficiency syndrome and neurodegenerative disorders. Apoptosis is characterized by cell shrinkage, membrane blebbing and nuclear condensation. To demonstrate the involvement of apoptosis in underlying mechanism for the development of silica-induced pathological changes, this study was designed in vitro and in vivo models. In in vitro study, alveolar epithelial cell line (A549) was stimulated with silica and performed flow cytometry and DNA electrophoresis. In in vivo study, bronchoalveolar lavage (BAL)was done to count the total and apoptotic cells from silica-instilled rats. The results were as follows: 1. Apoptotic cell fraction of silica-treated groups (10 and 50 ㎍/㎠) was significantly higher than that of control group. 2. Genomic DNA from silica-treated groups (10 and 50㎍/㎠) showed DNA ladder in agarose gel electrophoresis, while group of 1 ㎍/㎠ didn't. 3. Total cell number and apoptotic cell number of BAL fluid from silica-instilled rats (10, 20 and 40mg/kg) were significantly higher than those of control. 4. Silica induced apoptosis of cells in BAL fluid was confirmed by microscopic observation with nuclear fragmentation. These results suggest that apoptosis may contribute to development of silica-induced pathological changes.

제조업체 분진폭로 근로자들의 임상경과 및 폐기능 : Potential Biomarkers of Pneumoconiosis 진폐증에 관한 생체지표 개발

김지홍,정치경,장황신,안병용,박영만,임 영,김경아,구정완 大韓産業醫學會 1998 대한직업환경의학회지 Vol.10 No.4

Alveolitis is believed to be a primary response after dust inhalation. Activated inflammatory cells by dust and their mediators are major participants in the evolution of pneumoconiosis. Therefore evaluation of degree of alveolitis is useful for assessing disease activity and estimating of prognosis in pneumoconiosis. This study focused on the workers with pneumoconiosis who are working and worked in manufacturing industries because the occupying fraction by them among total pneumoconioese pateints in our country is increasing recently. In order to identify and validate biomarkers to early diagnosis and better predict for the suceptible workers, the release of tumor necrosis factor(TNFα), interleukine-8(IL-8), platelet-derived growth factor-AA(PDGF-AA) and transforming growth factor-β(TGFβ) from alveolar macrophages and blood monocytes, their concentration in BAL(bronchoalveolar lavage) fluid, pulmonary function test and Gallium index of lung were performed on 25 patients with pneumoconiosis who worked in manufactory industries. The results of this study were as follows; 1. No significant differences were demonstrated between large opacity group and small opacity group, however pulmonary diffusing capacity in large opacity group was significantly lower than that in small opacity group. 2. 67Gallium index was significantly correlated with increase of category in pnemoconiosis(r=0.83, p<0.05). 3. Numbers of total cells(r=0.583, p<0.05) and alveolar macrophages(r=0.499, p<0.05) in BAL were significantly correlated with the category of pnemoconiosis 4. Spontaneous(r=0.474, p<0.05) and LPS(r=0.463, p<0.05) stimulated release of TNFα from macrophage, IL-8(r=0.464, p<0.05) and TGFβ(r=0.460, p<0.05) in BAL fluid were significantly correlated with category of pnemoconiosis. 5. Significantly positive correlation was demonstrated between spontaneous(r=0.443, p<0.05) and LPS(r=0.573, p<0.05) stimulated release of TNFa with the category of pnemoconiosis. 6. Release of TNFα from alveolar macrophages(r=0.61, p<0.05) and blood monocytes(r=0.48, p<0.05), IL-8 concentration in BAL fluid(r=0.52, p<0.05) and 67Gallium index(r=0.53, p<0.05) were consider to sensitive biomarkers for alveolitis by dust.

단순진폐증에 대한 흉부 고해상 전산화 단층촬영의 진단적 의의

김경아,김지홍,장황신,안형숙,임영,윤임중,Kim, Kyoung-Ah,Kim, Hi-Hong,Chang, Hwang-Sin,Ahn, Hyeong-Sook,Lim, Young,Yun, Im-Goung 대한예방의학회 1996 Journal of Preventive Medicine and Public Health Vol.29 No.3

Early recognition of coalescence in pneumoconiotic lesions is important because such coalescence is associated with the respiratory symptoms and deterioration of lung function. This complicated form of pneumoconiosis also has worse prognosis than does simple pneumoconiosis. High resolution computerized tomography(HRCT) provides significant additional information on the stage of the pneumoconiosis because it easily detects coalescence of nodules and emphysema that may not be apparent on the simple radiograph. The purpose of this study is to clarify the role of HRCT in detection of large opacity and the relationship of change between the coalescence of nodules or emphysema and lung function in dust exposed workers. 1. There was good correlation between the HRCT grade of pneumoconiosis and ILO category of profusion. 5(9.09%) in 55 study population had confluent nodule extending eve, two o, more cuts on HRCT. HRCT could identify the pneumoconiotic nodules which was not found by simple radiogrphy in 6 workers with category 0/0. 2. No significant difference was observed coalescence of nodules and emphysema by dust type. 3. There was no significant difference in pulmonary function according to ILO and HRCT classification. 4. HRCT could detect the significant reduction in $FEV_1,\;FEV_1/FVC$, PEFR, $FEF_{25},\;FEF_{50},\;and\;FEF_{75}$ and remarkable increase in RV and TLC in study persons with emphysema compared with non-emphysema group. 5. Emphysema was found more often in nodules-coalescence group than small opacity group by HRCT. We found that HRCT could easily detect areas of coalescence and complicated emphysema compared to plain chest X-ray. Also our data suggest that it is primarily the degree of emphysema rather than the degree of pneumoconiosis that determines the level of pulmonary function.

광물성 섬유의 수산이온기 생성이 흰쥐 폐포대식세포의 세포독성과 적혈구 세포막의 지질과산화에 미치는 영향

김경아,윤임중,김지홍,장황신,오민화,임영,박정일 大韓産業醫學會 1997 대한직업환경의학회지 Vol.9 No.3

There is some evidence suggesting that the mechanism of pulmonary toxicity of natural fibrous silicate, asbestos, is related to the generation of oxygen-based free radical. Especially the hydroxyl radical(·OH) is believed to be the most reactive one. ·OH is a potent toxic oxidant, ·OH is which means highly cytotoxic, genotoxic and carcinogenic. Nevertheless there has been no previous report about ·OH generation from man-made mineral fibers except asbestos. In this paper we tested the hypothesis that both cytotoxicity of alveolar macrophage and lipid peroxidation of erythrocytes are associated with ·OH production by mineral fibers in vitro. The results were as follows: 1. ·OH production in vitro was progressively increased by the concentration dependent pattern with the same mineral fiber. The production of ·OH in vitro by man-made mineral fiber was markedly decreased compared with that of natural mineral fiber except rock wool. 2. Lipid peroxidation of mineral fiber-exposed erythrocyte suspension was significantly increased compared with that of phosphate-buffered saline(PBS) control group. Lipid peroxidation in amosite and chrysotile group was larger than that of other mineral fibers. 3. There was a significantly positive correlation between ·OH production in vitro and lipid peroxidation of mineral fiber-exposed erythrocyte. 4. All of mineral fibers were related to the cytotoxicity of alveolar macrophage in concentration-dependent fashion. Chrysotile and amosite were more cytotoxic than crocidolite. Among man-made mineral fibers, ceramic fiber was the most cytotoxic to alveolar macrophage. The viability of alveolar macrophage exposed to asbestos was markedly decreased compared with that of man-made mineral fiver at the same concentration. 5. There was the significant negative correlation between the production of .OH and the viability of alveolar macrophages. From these results, we can conclude that the production of ·OH in vitro was strongly correlated with the cytotoxicity of alveolar macrophage and the lipid peroxidation in mineral fiber-exposed erythrocyte. And we documented that man-made mineral fiber could generate .OH in vitro less than asbestos.

진폐증 치료제의 임상적 응용 및 개발에 관한 연구

김경아,임영,김주영,김지홍,장황신,박정일,윤임중 가톨릭대학산업의학센타 산업의학연구소 1998 韓國의 産業醫學 Vol.37 No.2

분진에서 장기간 폭로되면 분진의 종류에 따라 차이가 있지만 폐기능 저하와 함께 폐조직에 간질성 변화가 발생하는데 이를 흔히 진폐증이라 한다. 진폐증의 역사는 광산의 역사라고 할만큼 광산 근로자와 밀접한 관계가 있음으로 광산업이 퇴조하고 있는 우리나라에서 진폐증은 사라져 가고 있다고 흔히 생각하기 쉽다. 우리나라에서도 과거에는 거의 모든 진폐증이 광산근로자에서 발생되었으나 근래에는 각종 제조업체의 분진근로자, 건설업체의 근로자 뿐 아니라 분진 작업량 주위의 일반 주민에서도 발생된바 있어(윤임중등, 1994) 사회문제로까지 확대된 바 있다. 광산은 밀폐되고 환기가 잘 안되는 작업환경과 높은 분진농도 때문에 진폐증의 발생율이 높으나 산업구조의 변화에 따른 광산업의 퇴조로 광산근로자에서의 진폐증 발생이 감소하고 있으며 상대적으로 제조업체나 건설업에서의 진폐증 발생율이 증가하고 있다. 그러나 제조업체에서의 진폐증 발생율은 광산업에 비하여 낮아서 과거 우리나라 직업병 발생율의 70%에 달하여 가장 높은 비율을 차지하였던 진폐증이 50% 이하로 감소하긴 하였으나 여전히 높은 발생율을 유지하고 있다. 더욱이 진폐증은 비가역적이고 진행성의 성향을 가지고 있으며 합병증 발생이 빈번하여 계속적인 치료를 요하는 환자가 많다. 또한 현재 의사진폐증이나 소음영진폐증으로 장해대상이 되지 않아 관찰중인 비교적 경한 진폐증 환자도 대음영 진폐증으로 진행할 수 있으며 합병증 발생으로 치료가 필요할 수 있다. 실제로 1997년 3월 현재 전국의 19개 진폐관리병원에서 약 2,500명에 달하는 진폐증 환자가 폐기능 장해와 합병증으로 치료받고 있으며 지금까지 진폐증으로 장해보상을 받은 환자는 648명, 그리고 1966년부터 현재까지 진폐증으로 사망한 환자가 3,114명에 달한다. 그리고 매년 진폐정밀진단 검사상 진폐증은 있으나 장해대상이 안된다고 판정받는 환자수가 1,000여명에 이르며 의사진폐증으로 진단받은 경우도 수백명에 이르는데 진폐증이 비가역적이고 진행성의 질환임을 감안할 때 이들에 대한 관리 및 치료 그리고 더 이상의 진행을 막는 것이 근로자의 보건뿐 아니라 국가적인 차원에서 진단, 치료 및 장해보상을 시행하고 있음으로 국가적인 자원소실을 방지하는 차원에서도 중요한 의미를 갖는다. 그러나 전세계적으로 그리고 우리 연구소에서도 진폐증을 치료하는 방법을 개발하고자 오랜 기간동안 많은 노력을 기울렸으나 진폐증의 치료법은 아직까지 알려지지 않은 상태이다. 이처럼 진폐증의 치료제 개발이 어려웠던 원인으로는 진폐증의 병리기전이 명확하게 밝혀지지 않았기 때문인데 진폐증은 매우 오랜 역사를 가지고 있음에도 불구하고 분진 폭로에 의한 간질성 폐질환이 하기도의 만성적인 염증반응에 의한 것으로 알려진 것은 최근 10여년 간의 연구 결과이다 (Warheit 등, 1986). 따라서 진폐증 치료제의 개발은 진폐증의 발생 및 진행기전에 대한 규명이 선행되어야 하고 간질의 섬유화라는 최종 결과가 나타나기 전의 중간단계에서 질환의 발생을 억제할 수 있는 방법을 찾아내는 것이 타당할 것이다. 지금까지의 진폐증의 발생 및 진행기전이나 치료효과에 관한 연구들은 유리규산분진을 중심으로 진행되어 왔다. 그러나 산업이 발달함에 따라 제조업체에서 사용되는 분진의 종류도 다양해지고 있는데 폐장의 조직변화나 병리기전은 분진의 종류에 따라 다를 것으로 생각되고 있다. 특히 20세기 들어 산업화되면서 다량으로 사용되었던 석면은 자연적으로 존재하는 섬유형 분진으로 강력한 섬유화성 및 발암성이 특징적인데 석면의 발암성은 섬유형 분진의 모양이나 내구성과 연관되어 설명되기도 한다(Lippman등, 1994). 석면의 독성이 알려진 이후 개발되어 사용되는 있는 인조섬유들은 석면과 같은 섬유형 분진이며 이들에 대한 안정성 여부는 아직까지 확실하지 않은 상태이다. 이에 연구자들은 유리규산분진을 비롯하여 최근 제조업체나 건설현장에서 자주 이용되는 인조섬유에서 진폐증의 병리기전에 관여하는 여러 가지 미개물질이나 산소유리기의 관련성에 관한 연구들을 시행하여 진폐증의 병리 기전을 밝히고 치료방법을 개발하고자 1994년 한국학술진흥재단의 후원으로 3년간 연구를 실시한 바 있으며 이에 대한 결과를 종합하여 기술하고자 한다.

광물성 섬유의 폐장내 주입에 따른 조직변화와 MIP-2의 관련성

윤임중,김지홍,김경아,김흥남,이동원,박정일,장황신,임영,서화석 大韓産業醫學會 1997 대한직업환경의학회지 Vol.9 No.4

The pulmonary recruitment and activation of inflammatory cells, in particular, neutrophils is thought to contribute to lung injury resulting from dust exposure. MIP-2(macrophage inflammatory protein-2) which is a member of C-X-C chemokine plays a key role in neutrophil recruitment to sites of tissue injury. Especially, mineral fiber induced pulmonary response is as a model for the neutrophil recruitment. Therefore, we evaluated the distribution of MIP-2 expression in lung tissue of mineral fiber exposed rat using immunohistochemical study and the relationship between degree of inflammation of lower respiratory tract and MIP-2 expression. Total cell counts in bronchoalveolar lavage(BAL) fluid in mineral fiber-exposed group were markedly increased compared with each control group even not in ceramic fiber group. Number of neutrophil in BAL fluid in mineral fiber-exposed group were markedly increased compared with each control group until 4th weeks but except ceramic fiber group. In chrysotile group, number of neutrophil in BAL fluid were markedly increased compared with control group at 8the week. Lung tissue instilled with all kinds of mineral fibers showed remarkable developments of bronchus associated lymphoid tissue(BALT) and small multiple granulomas but not for ceramic fiber group. In chrysotile group, multiple granuloma and inflammatory change were more profuse response compared with other groups. MIP-2 was predominently expressed in epithelial cells of bronchioles and bronchus and was express also found in macrophages with lung section at 1 week after fiber instillation. Small amount of epithelial cell associated MIP-2 was present in chrysotile at 8 week group. But MIP-2 was not seen in epithelial cells and macrophages in the lung tissue instilled with crocidolite, ceramic fiber and glass fiber at 8 weeks. Our finding suggest that MIP-2 is predominantly expressed in bronchial epithelial cells of lung from mineral fiber-exposed rat and correlated with inflammatory cell, especially neutrophil, recruitment and tissue reaction. And we documented that MIP-2 expression and neutrophil recruitment in man-made vitreous fiber-exposed rat, especially glass fiber, less than chrysotile.

진폐증에서 비전형적 형태의 진행성 종괴성 섬유증

안병용 ( Byoung Yong Ahn ),박영만 ( Young Mann Baak ),장황신 ( Hwang Shin Chang ),김지홍 ( Jee Hong Kim ),김경아 ( Kyoung Ah Kim ),임영 ( Young Lim ) 대한결핵 및 호흡기학회 1999 Tuberculosis and Respiratory Diseases Vol.47 No.2

진폐증 환자에서의 혈청내 IL-8 농도

안형숙 ( Kyeong Sook Ahn ),김지홍 ( Ji Hong Kim ),장황신 ( Hwang Sin Chang ),김경아 ( Kyung Ah Kim ),임영 ( Young Lim ) 대한결핵 및 호흡기학회 1996 Tuberculosis and Respiratory Diseases Vol.43 No.6