천식유발 마우스에서의 폐 내 세포조성 변화와 IL-4 및 IL-10의 발현 양상

이수진,박세종,리천주,장양호,최농훈,Lee, Soo-Jin,Park, Se-Jong,Li, Tian-Zhu,Jang, Yang-Ho,Choe, Nong-Hoon 한국생명과학회 2006 생명과학회지 Vol.16 No.5

본 연구진은 일차로 BALB/C 마우스를 이용하여 항원으로 ovalbumin을 사용하여 천식을 유발하여 폐 내 세포구성에 미치는 영향을 다른 실험방법에 의해 만들어진 천식모델과 비교하여 살펴보았고 이차로 천식 발생에 중요한 역할을 하는 것으로 알려진 IL-4와 IL-10 knock out (KO) 마우스를 이용하여 천식모델을 구축하여 천식인자를 가진 개체에서의 반응과 천식관련인자가 결핍된 개체에서의 반응성의 차이를 살펴보았다. 천식의 유발은 실험 1일째 $20\;{\mu}g$ ovalbumin으로 감작시킨 후 실험 14일에 재감작을 시켰다. 그 후 nebulizer를 이용하여 nasal inhalation을 28일, 29일, 30일에 실시하여 천식을 유발시켰다. 천식 유발의 확인은 기관지폐포 세척술로 채취한 폐 내 세포액을 이용하여 총 세포수 및 염증세포의 증가와 폐 내 세포와 폐조직의 염색을 통해 분포율을 확인하였다. 천식의 발병 과정에서 IgE 관련 과민반응을 주도하는 IL-4와 Th2 세포의 기능을 억제한다고 알려진 IL-10의 면역화학염색을 통해 그 발현정도를 관찰하였다. BALB/c 마우스의 천식유발군의 경우 천식의 특징인 염증세포의 증가와 호산구의 증가와 세기관지 주위 염증세포의 침윤 및 기도 상피의 비후를 관찰할 수 있었다. 따라서 본 실험에 응용된 천식유발 방법은 적합하였으며 천식유발 절차가 성공적으로 수행되었음을 확인하였다. IL-4 및 IL-10 KO 마우스를 이용한 천식유발군의 경우 BALB/c 마우스를 이용한 천식유발군보다 호산구의 증가 정도가 미약하게 관찰되었고, 폐조직 내 염증세포의 침윤 정도도 감소하였다. 천식매개인자인 IL-4와 IL-10의 면역조직화학염색 결과에서도 양성반응이 거의 나타나지 않았다. 본 연구 결과, IL-10이 IL-4와 같이 천식과 관련하여 세기관지 염증 반응을 증가시키는 역할을 하는 것으로 생각된다. Asthma is a chronic inflammatory disorder of the airways, which characterized by bronchial hyperresponsiveness, reversible airflow limitation and respiratory symptoms. Internationally, the prevalence of asthma has been increased over last 3 decades. Recently, several studies of asthma have been reported with gradually increasing importance. To tesify the hypothesis that interleukin (IL)-4 and IL-10 may be an important determinant of the severity of airway inflammation, their expression was studied in mouse model of asthma. BALB/c mouse, IL-4 Knockout (KO) mouse and IL-10 KO mouse were sensitized with intraperitoneal injection of ovalbumin adsorbed to aluminum potassium sulfate, followed by challenges with intranasal ovalbumin on 3 consecutive days. The severity of pulmonary inflammation was assessed by eosinophilia in BAL fluid, number of total BAL cells, histopathological changes in lung tissues, and immunohistochemical staining against IL-4 and IL-10. In BAL fluid, the number of total cells was significantly increased in asthma induced mouse compare to the control. In asthma induced mouse, eosinophil was increased to 56% and neutrophil was 0.2%. In H &E stains, eosinophilic infiltration and epithelium hyperplasia were clearly noticed in asthma induced mouse. In immunohistochemical staining for IL-4 and IL-10, there was no positive reaction in control group. However, very strong reactions were appeared in asthma induced group. In this research, IL-4 and IL-10, which seem to play a central role in allergic asthma, KO mouse was utilized to test the causative relationship between airway inflammation and role of specific cytokine. Asthma induced IL-4 and IL-10 KO mice showed much decreased inflammatory reactions in the number of total BAL cells, in eosinophilic infiltration, and in immunohistochemical stains against diverse inflammatory proteins. These results suggest that IL-4 and IL-10 increase the asthmatic reactions in vivo mice model.

랫드 궁둥신경에서 155 kd Neurofascin의 발현개시에 대한 면역세포화학적연구

원희영,조익현,이종환,남상섭,최농훈,박재형,장병준,Won, Hee-Young,Cho, Ik-Hyun,Lee, Jong-Hwan,Nahm, Sang-Soep,Choe, Nong-Hoon,Park, Jae-Hyung,Chang, Byung-Joon 대한수의학회 2012 大韓獸醫學會誌 Vol.52 No.3

Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the development of nerve fibers in the rat sciatic nerve, the initial development of NF155 in the paranode was studied with immuno-fluorescence and immuno-electron microscopy. The result of the present study showed NF155 was not detected in the fetal sciatic nerve and began to reveal at the postnatal day 0 (P0) and dramatically increased by time lapse until postnatal day 7 (P7). NF155 was prominently localized in the axolemma of paranode and not detected in the central region of node of Ranvier. According to the present study, NF155 is likely to have some relationships with the formation of paranode and myelin sheath.

디젤분진 및 미세분진이 천식마우스에서 기도 재구성에 미치는 효과

리천주,이수진,장양호,박준홍,박세종,이정학,최농훈,Li, Tianzhu,Lee, Soo-Jin,Jang, Yang-Ho,Park, Jun-Hong,Park, Se-Jong,Lee, Jeong-Hak,Choe, Nong-Hoon 한국생명과학회 2007 생명과학회지 Vol.17 No.2

천식이 유발된 Balb/c 마우스와 동일한 조건의 Il-10 KO 마우스에 천식의 원인으로 알려진 DEP와 지하철역내에서 채칩한 PM ($10mg/m^{3}$)을 inhalation chamber에 넣고 하루 4시간씩 5일간 흡입시킨 후 시료를 채취하여 기관지 상피하부의 섬유화정도와 술잔세포의 증식 정도를 살펴 천식증상의 악화에 DEP와 PM이 어떠한 영향을 미치는지 확인하였다. 본 실험의 결과 천식이 유발된 일반 Balb/c 마우스에 있어서는 DEP와 PM의 노출에 의하여 상피하부의 섬유화가 증가하였고, 술잔세포의 수도 증가함을 확인하였다. 그러나, Il-KO 마우스의 경우 상피하부의 섬유화정도와 술잔세포의 수가 거의 증가하지 않았다. 따라서, 본 결과는 IL-10에 대한 항체요법이 천식증상의 완화에 쓰일 수 있는 가능성을 암시하며, 한편 자동차 배기가스와 지하철 미세분진의 발생을 예방할 경우 천식과 관련한 세기관지의 염증을 완화시킬 수 있음을 간접적으로 증명한 것이라 할 수 있다. This research investigated whether exposure of diesel exhaust particulate (DEP) and particulate matter (PM) effects on airway remodeling in asthma induced Balb/c and IL-10 knock out (KO) mouse. Mice were sensitized with intraperitoneal injection with ovalbumin, followed by challenges with intranasal ovalbumin. After that mice placed in inhalation chamber and exposed to DEP and $PM(10\;mg/m^3)$. The evidence of airway remodeling was assessed by masson's trichrome staining and PAS staining. The stainability of masson's trichrome and PAS reaction were increased in asthma-induced Baltic mice groups compared with control mice groups. More intensive stainability for masson's trichrome and PAS were appeared in the asthma-induced DEP and PM-exposed groups than asthama-induced groups. But, not significantly increased subepithelial fibrosis and the nember of goblet cell hyperplasia in asthma-induced IL-10 KO mice groups and asthma-induced+DEP and PM-exposed IL-10 KO mice than IL-10 KO mice groups. These results indirectly suggesting that exposure to DEP and PM in asthmatic patients might be aggravate clinical symptoms and IL-10 which seems to play a central role in allergic asthma. In conclusion, DEP and PM exposure might have additive effects on the ovalbumin- induced asthma in a murine model.

Active biotelemetry를 이용한 젖소의 스트레스 반응 측정 I.보정 스트레스에 대한 경산우와 미경산우의 생체반응의 변화

임좌진,이병한,김진영,이수한,이동희,김재경,정순욱,최농훈,정병현,Lim, Joa-jin,Lee, Byeong-han,Kim, Jin-young,Lee, Su-han,Lee, Dong-hee,Kim, Jae-kyung,Jeong, Soon-wuk,Choe, Nong-hoon,Chung, Byung-hyun 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.4

This study was carried out to investigate the effectiveness of active biotelemetry to monitor stress response, and the effects of acute stressor (raising a hind foot of a standing animal) on body temperature (BT) and heart rate (HR) in comparison heifer group (n=4) with cow group (n=3). Both parameters were recorded by active telemetry. Cortisol assay was used to compare the result of active biotelemetry. The stressor consisted in restraining the cows for hoof treatments. Body temperature and heart rate were measured at 0, 10, 20, 30, 40, 50, 60, 90, 120, 180 and 360 minutes during the experiments, and also cortisol assay at 0, 0.5, 1, 2, 3, 12 and 24 hours. Base line for these animals was set in there accustomed environment at -24hrs and -30 mins before the beginning of experiments. The results obtained in this study were as follows: 1. In the BT, heifer group showed no significant difference from cow group, but were observed slightly higher BT than cow group during restraint stress for 30 minutes. 2. The both groups showed directly significant increase of HR immediately after the beginning of the restraint stress (P<0.01), but in the required time for the recovery of HR after finishing the stress, heifer group was 40 minutes and cow group 60 minutes. 3. In the cortisol assay, the both groups increased from 30 to 120 minutes after the beginning of experiment (i.e, from 0 to 90 minutes after finishing the restraint stress), showing significant difference from before the beginning of the stress (P<0.01), but heifer group showed great increase of the cortisol level from 30 to 60 minutes after the beginning of the stress, while cow group was observed at 60 minutes. In conclusion, we suppose that care is required to decrease psychological stress in cattle when they suffer from a series of procedures for diagnosis and treatment, especially in heifers rather than in cows. Also BT and HR obtained by active biotelemetry are reliable indicators of stress in cattle.

말초신경의 재생에 대한 녹용의 효과; 랫드에서 실험적 절단 좌골신경의 Sprout 형성에 관해

장병준,조익현,최혜영,원희영,박창현,배춘식,최농훈,Chang, Byung-Joon,Cho, Ik-Hyun,Choi, Hye-Young,Won, Hui-Young,Park, Chang-Hyun,Bae, Chun-Sik,Choe, Nong-Hoon 한국현미경학회 2002 Applied microscopy Vol.32 No.1

좌골신경 재생에 미치는 녹용의 효과를 알아보기 위해 300 gm 내외의 수컷 Sprague-Dawley계 랫드에 체중 100 gm당 1.5 ml의 수용성 녹용추출물을 각각 1주, 2주, 3주간 매일 1회씩 경구 투여한 후 양쪽 하지의 좌골신경을 절단하였다. 6시간 경과 후 절단부의 근위부에서 절취한 좌골신경을 통상적인 전자현미경기법으로 처리하여 관찰한 바 다음과 같은 결과를 얻었다. 1. 정상대조군의 좌골신경은 신경마디에서 axonal sprouting이 관찰되지 않았고, 전자밀도가 높은 축삭막의 돌출구조가 종종 관찰되었다. 2. 생리식염수 투여군의 좌골신경의 신경마디에서 axonal sprouts의 형성이 관찰되었으며, 그 길이는 대부분 짧았고, 내부에 많은 소포와 공포를 함유하고 있었으며 일부에서는 신경세사를 비롯한 세포소기관을 가지고 있었다. 각각 100개씩의 세로절단된 신경마디에서 sprouts 의 존재여부를 계수한 바 1주 투여군에서 29개(29%), 2주 투여군에서 32개 (32%), 3주 투여군에서 30개 (30%)로 각각 나타났다. 3. 녹용 투여군의 좌골신경의 신경마디에서 axonal sprouts의 형성이 관찰되었고 대부분 그 길이가 짧았으나 2주 투여군과 3주 투여군에서 종종 긴 sprouts가 관찰되었다. Sprouts의 내부에는 많은 소포와 공포를 함유하고 있었고 종종 신경세사, 사립체 등의 소기관을 함유하고 있는 경우도 있었다. 각각 100개씩의 세로절단된 신경마디에서 계수한 sprouts의 수는 1주 투여군에서 38개 (38%), 2주 투여군에서 46개 (46%), 3주 투여군에서 48개(48%)로 각각 나타났다. 이상의 결과로부터 녹용추출물의 선투여는 절단좌골신경의 sprout 형성을 촉진함을 알 수 있었고, 이는 녹용이 말초신경의 재생에 효과가 있음을 시사한다고 생각된다.와 전자밀도가 낮은 소포들이 혼합된 형과 전자밀도가 낮은 소포들로만 이루어진 연접체 등 두 종류를 소지하고 있어 두 종간 차이가 확인되었다.흰쥐 전뇌 기저부 Meynert기저핵에서의 ChAT면역반응 신경세포들은 출생 후 발생과정에서 세포소기관과 신경돌기들의 분화에 따른 세포체 부피의 증가, 분화된 세포형들의 출현 율의 증가 및 세포의 손실이 없는 상태에서 세포 응축 등의 과정을 통하여 세포들이 분화한다고 생각된다.\mu}m$정도로 세포질 대비 매우 컸다. 이들은 다양한 크기의 전자밀도가 높은 둥근 과립(직경 $1\sim0.2{\mu}m$)을 소지하고 있었으며, 세포의 표면은 여러 형태의 사상족(filopodia)들을 뻗어 노쇠한 세포들을 포식하였다. 신경망(neuropiles)들은 신경분비세포를 둘러싸고 있었으며, 신경섬유 속에서 다양한 종류의 연접소포들(synaptic vesicles)이 관찰되었는데, 전자밀도, 크기 그리고 모양에 따라 6종류로 분류되었다.로 확인되었다. 방사대는 외측에 않은 homogeneous layer와 내측에 2개의 두꺼운 helicoidal layers (zona radiata interna and zona radials externa)총 3개의 구조물로 구성되어 있었다. 정상적인 환경조건에서 볼 때 난포는 난형성과 생식소에서 분비되는 스테로이드 흐르몬의 생성에 충분할 만큼 조직의 발달과 호르몬의 주기적인 분비에 영향을 주는 것으로 나타났다.%o보다 20%o, 30%o에 적응(適應)된 개체의 신장조직(腎臟組織)에서 더많이 발생되었다. 틸라피아의 신사구체(腎絲球體)는 담수(淡水)에서 10%o의 해수(海水)로 이주된지 14일(日) 이후에 신장(腎臟)에서 수축된 것으로 나타났다. 30%o의 해수(海水)에 적응(適應)된 틸라피아의 평균 신사구체(腎絲球體)의 면적은 This study was carried out to investigate the effects of deer antler extract on the regeneration of peripheral nerves. Sprague-Dawley male rats weighing about 300 gm were fed deer antler extract for 1, 2, and 3 weeks per oral (1.5 ml/100 gm B.W.), respectively, once a day and transected both sides of sciatic nerve of each leg. After keeping for 6 hours, sciatic nerves taken from proximal part of transected region were treated with conventional transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows; 1. Sciatic nerves of normal control group were not showing any sprouts and electron dense axolemmal projections were frequently observed. 2. Sciatic nerves of saline treated groups were showing axonal sprouts at the nodes of Ranvier. The length of them was usually short, and numerous vesicles, vacuoles and organelles including neurofilament were contained. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 29 (29%) in 1 week treated group, 32 (32%) in 2 weeks treated group, and 30 (30%) in 3 weeks treated group, respectively. 3. Sciatic nerves of deer antler treated groups were showing axonal sprouts at the node of Ranvier as well. Although most of the sprouts were short, some sprouts of 2 weeks and 3 weeks treated groups were quite long. Sprouts usually contained numerous vesicles, vacuoles and cell organelles such as neurofilaments and mitochondria. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 38 (38%) in 1 week treated group, 46 (46%) in 2 weeks treated group, and 48 (48%) in 3 weeks treated group respectively. The results described above explain pretreatment of deer antler extract improves the sprout formation of transected sciatic nerves, and then it suggests deer antler may be effective for the regeneration of peripheral nerves.

랫드 수초좌골신경섬유에서 Neurofascin분포에 대한 면역세포화학적 연구

장병화,유관희,이종환,조익현,배춘식,박창현,한정미,최농훈,장병준,Chang, Byung-Hwa,You, Kwan-Hee,Lee, Jong-Hwan,Cho, Ik-Hyun,Bae, Chun-Sik,Park, Chang-Hyun,Han, Jeong-Mi,Choe, Nong-Hoon,Chang, Byung-Joon 한국현미경학회 2006 Applied microscopy Vol.36 No.2

Neurofascin은 L1CAM의 하나로 신경섬유의 발달과정에서 중요한 역할을 하는 것으로 알려져 있다. 말초신경의 수초형성과 관련된 neurofascin의 역할을 알아볼 목적으로 면역형광염색과 면역전자현미경기법을 이용하여 랫드의 수초좌골신경섬유에서 neurofascin의 분포를 추구하여 다음과 같은 결과를 얻었다. 1.수초형성이 진행됨에 따라 좌골신경섬유에서 neurofascin 분포는 매우 심하게 변화되었다. 2. 수초신경섬유에서 neurofascin은 Ranvier마디에서 약하게 국재하였다. 3. Neurofascin은 수초신경섬유의 paranodal loop, Schmidt-Lantermann incisure, 속축삭사이막, 바깥축삭사이막처럼 Schwann세포의 막이 밀착되지 않은 부위에서도 뚜렷하게 국재하였다. 이상의 연구결과로 neurofascin은 Schwann세포의 마주보는 막사이에 이상적인 간격을 유지하는데 어떤 역할을 하는 것으로 생각되며, 수초층에서 물질이동이 가능하게 하는 것으로 보인다. Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the myelination of peripheral nerves, the localization of neurofascin in myelinated rat sciatic nerve fibers was studied with the immuno-fluorescence and immune-electron microscopy and the results are as follows; 1. According to the myelination is going on, neurofascin localization was dramatically changed in the sciatic nerve fibers. 2. In the myelinated fibers, neurofascin was weakly localized along the axolemma at the node of Ranvier. 3. Neurofascin was also apparantly localized at the non-compact area of Schwann cell membrane such as paranodal loop, Schmidt-Lantermann incisure, inner & outer mesaxons in the myelinated fibers. From the above results, neurofascin is likely to have a role to sustain the ideal gap of apposing membranes of Schwann cell, so it may enable to materials transport in the myelin sheath.

국내 흑염소에서의 소결핵, 브루셀라, Q fever 유병률 연구

김효비,김성준,김기나,김별,장병준,최농훈,Kim, Hyobi,Kim, Seongjoon,Kim, Kina,Kim, Byeol,Chang, Byungjoon,Choe, Nong-Hoon 대한수의학회 2016 大韓獸醫學會誌 Vol.56 No.4

As the meat of black goats has become popular as a healthy food, domestic goat meat-related industries are steadily growing. However, previous studies are scarce of informations about the zoonotic disease originated from the black goat in Korea. In this study, we investigated Korean black goat's infectious diseases representing bovine tuberculosis, brucellosis, and Q fever. One hundred and eighty samples were collected from a local slaughter house located in Jeollanam-do. Three typical zoonotic diseases were separately examined by carrying out enzyme linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Histopathological test was additionally performed in tuberculosis. In case of tuberculosis, results of the PCR and histopathological test were negative but the ELISA results were positive in eight samples. In case of brucellosis, one out of the total samples was shown to be positive in the ELISA and none in the PCR. In case of Q fever, there were forty one positive in the ELISA and twenty positive in the real-time PCR. Those results indicate that the Korean black goat could be a natural reservoir in the possible chain-infections among human, cows and goats. Thus, further study needs in order to improve productivity as well as to prevent the zoonosis spreading and circulation of other livestock with the black goat in this country.

Active biotelemetry를 이용한 젖소의 스트레스 반응 측정: II. 수송 스트레스 감소를 위한 수액투여 효과

이수한,이병한,임좌진,김진영,이동희,김재경,최농훈,정순욱,정병현,Lee, Su-han,Lee, Byeong-han,Lim, Joa-jin,Kim, Jin-young,Lee, Dong-hee,Kim, Jae-kyung,Choe, Nong-hoon,Jeong, Soon-wuk,Chung, Byung-hyun 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.4

A transport stress is one of the main causes of economic losses and physiological dysfunction. The present study has been performed to suggest a method to decrease the adverse effects above mentioned from transport. The groups were prepared as follows; (1) Control group : 4 cattle transported for 5 hrs (274 km) without any treatment, (2) Treatment group : 4 cattle treated with electrolyte-mineral solution (I.V.) at 1 hr before the enrollment of transport under same experimental condition with Control group. The blood specimens were collected at 1 hr before transport, 2.3 hrs (135 km) and 5 hrs (274 km) after the enrollment of transport, and 1, 6 and 18 hrs after fulfillment of transport. The collected blood specimens were analyzed for cortisol and epinephrine. Core temperature and heart rate were measured with active biotelemetry in every 30 minutes from 0.5 hr before the start of transport to 18 hrs after the end of transport. In results, the level of cortisol considerably increased to the peak either in Control group ($5.3{\pm}1.3{\mu}g/d{\ell}$) and in Treatment group ($4.0{\pm}2.6{\mu}g/d{\ell}$) at 2.3 hrs in transport. The concentration of epinephrine of Treatment group had been higher than that of Control group from the start of transport to 18 hrs after the fulfillment of transport. Particularly there was the biggest gap between Control and Treatment groups, $424.0{\pm}194.1pg/m{\ell}$ and $209.1{\pm}65.1pg/m{\ell}$ respectively, at 6 hrs after the end of transport. The heart rates were considerably increased either in Control group ($81.5{\pm}18.5$ to $126.3{\pm}7.8beats/min$) and in Treatment group ($114.3{\pm}14.4$ to $140.8{\pm}22.4beats/min$) with the enrollment of transport. These results indicate that the concentration of cortisol and the heart rate were pertinent to cognitive parameters to evaluate physiological responses against stress such as transport. In addition, the intravenous administration of electrolyte-mineral solution could be suggested as the method to decrease the adverse effects from a transport stress.

디젤분진 및 미세분진이 천식마우스의 폐조직에서 ICAM-1과 VCAM-1의 발현에 미치는 효과

리천주,이수진,장양호,이정학,박세종,박준홍,장병준,이종환,최농훈,Li, Tian-Zhu,Lee, Soo-Jin,Jang, Yang-Ho,Lee, Jeong-Hak,Park, Se-Jong,Park, Jun-Hong,Chang, Byung-Joon,Lee, Jong-Hwan,Choe, Nong-Hoon 한국생명과학회 2007 생명과학회지 Vol.17 No.3

천식이 유발된 Balb/c마우스와 동일한 조건의 IL-10 KO 마우스에 천식의 원인으로 알려진 DEP와 지하철역내에서 채집한 PM (10 ${\mu}g/m^3$)을 inhalation chamber,에 넣고 하루 4시간씩 흡입시킨 후 시료들을 채취하여 ICAM-1, VCAM-1의발현을 살펴 천식증상의 악화에 DEP와 PM이 어데한 영향을 미치는지 확인하였다. 본 실험의 결과 천식이 유발된 일반 Balb/c 마우스에 있어서는 DEP와 PM의 노출에 의하여 ICAM-1 및 VCAM-1의 발현이 세기관지 주위 조직들에서 미약하게 증가하였다. 그러나 IL-10 KO 마우스의 경우 DEP와 PM을 노출시켰을 때 ICAM-1 및 VCAM-1의 발현이 아주 강하게 증가하였다. 따라서, 본 결과는 IL-10에 대한 항체요법이 천식증상의 완화에 쓰일 수 있는 가능성을 암시하며, 한편 자동차 배기가스와 지하철 미세분진의 발생을 예방할 경우 천식과 관련한 세기관지의 염증을 완화시킬 수 있음을 간접적으로 증명한 것이라 할 수 있다. This research investigated whether exposure of diesel exhaust particulate (DEP) and particulate metter (PM) effect on intercellular. adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression in asthma-induced Balb/c and IL-10 knock out (KO) mouse. Mouse was sensitized with intraperitoneal injection with ovalbumin, followed by challenges with intranasal ovalbumin. After induction of asthma mouse placed in the inhalation chamber and exposed to DEP and PM (10 $mg/m^3$). The evidences of pulmonary inflammation were assessed by immunohistochemical stain and westen blot against ICAM-1 and VCAM-1 in the lung tissue. In the immunohistochemical stain, positive reactions for ICAM-1 and VCAM-1 were much stronger in asthma-induced groups and asthma-induced group with DEP or PM than control groups. Although mild positive reactions were appeared in asthma-induced IL-10 KO mice groups, positive reactions were very strong in the asthma-induced group with DEP or PM. In Western blot, expression of VCAM-1 was increased in asthma-induced group with DEP or PM than asthma-induced groups. In the IL-10 KO mouse, ICAM-1 and VCAM-1 expression were increased in asthma-induced group with DEP or PM than asthma-induced groups. DEP and PM exposure have additive effects on the aggravation of inflammatory signs in the asthma-induced murine model. These results suggest that inhalation of DEP and PM in asthmatic patients may aggravate clinical symptoms.

디젤분진이 폐포대식세포에서 nitric oxide의 생성과 inducible nitric oxide synthase의 발현 및 nitrotyrosilated-protein의 형성에 미치는 효과

임영,최명옥,이권행,김경아,김길수,이명헌,리천주,이수진,최농훈,Lim Young,Choe Myung-Ok,Lee Kweon-Haeng,Kim Kyung-A,Kim Kil-Soo,Lee Myoung-Heon,Li Tian-Zhu,Lee Soo-Jin,Choe Nong-Hoon 한국생명과학회 2006 생명과학회지 Vol.16 No.2

본 연구에서는 DEP의 노출이 새로운 호흡기계 질환 유발의 가능성과 호흡기계의 염증성인자로 잘 알려진 lipopolysaccharide (LPS)의 역할에 어떠한 영향을 미치는 지를 확인하고자 폐에서 염증성 반응 시 생성이 증가하는 것으로 알려진 Nitric Oxide (NO)의 형성과 NO의 생성에 관련된 효소인 inducible nitric oxide synthase (iNOS) 및 NO에 의하여 형성되는 것으로 알려진 nitrotyrosilated-protein을 폐포 대식세포를 통해 분석하였다. 폐포대식세포에 DEP를 농도 별로 단독 처리하였을 때와 동일한 농도에서 배양시간을 달리하였을 때는 NO가 생성되지 않았으나 DEP와 함께 LPS를 처리하였을 때는 LPS를 단독으로 처리했을 때보다. 유의성이 있게 증가함을 확인할 수 있었다. 또한 NO의 생성에 관련된 효소인 iNOS 및 NO에 의하여 형성되는 것으로 알려진 nitrotyrosilated-protein 발현의 정도를 면역화학염색과 Western analysis로 확인할 수 있었다. DEP는 폐포대식세포에서 직접적으로 NO생성에 영향을 미치지 않았으며, NO를 생성하는 iNOS나 nitrotyrosilated-protein의 발현에도 영향을 주지 않았으나 세균성 염증인자의 한 종류인 LPS가 NO를 형성하는 데에는 통계학적인 상승효과가 있었다. 결론적으로 본 연구에서는 염증성질환의 환자에서 DEP의 흡입은 간접적으로 NO를 형성하는데 영향을 미쳐 질환을 악화시킬 것으로 판단한다. Epidemiological studies have demonstrated an association between exposure to diesel exhaust particles (DEP) and adverse cardiopulmonary effects. Despite the epidemiological proof, the pathogenesis of DEP-related pulmonary diseases remain poorly understood. So, comprehensive in vivo and in vitro researches are required to know the effects of DEP on diverse lung diseases. Alveolar macrophages (AM) and airway epithelial cells are known as important cellular targets in DEP-induced lung diseases. Other studies have shown that nitric oxide (NO) is involved in particle matter induced lung injury. The present study was undertaken to determine whether DEP has an synergistic effects on lipopolysaccharide (LPS)-induced NO formation and inducible nitric oxide synthase (iNOS) with nitrotyrosilated-protein formation in cultured primary alveolar macrophages. The formation of NO was determined through the Griess reaction in the cultured medium and iNOS with nitrotyrosilated-proteins are analyzed by immunohistochemical staining and Western analysis. The results indicate that DEP exposure does not induce NO formation by itself, however DEP showed significant synergistic effects on LPS-induced NO formation. So, our results suggest that DEP inhalation could aggravate inflammatory lung disease through NO formation.