Alteration of PMN Leukocyte Function by the Change of Sulfhydryl Group and Metabolism of Membrane Components

신재훈,이정수,한은숙,신용규,이광수,Shin, Jeh-Hoon,Lee, Chung-Soo,Han, Eun-Sook,Shin, Yong-Kyoo,Lee, Kwang-Soo The Korean Society of Pharmacology 1989 대한약리학잡지 Vol.25 No.1

면역 보체가 결합되어 있는 zymosan에 의하여 활성화된 다형핵 백혈구에서 세포 투과성 물질인 N-ethylmaleiamide과 $Hg^{++}$은 superoxide 라디칼 생성, NADPH oxidase 활성도 및 lysosomal enzyme (lactic dehydrogenase, ${\beta}-glucuronidase$)의 유리를 억제하였다. 세포막 단백에 특이적인 p-chloromercuribenzoic acid와 p-chloromercuribenzenesulfonic acid는 superoxide 라디칼 생성에 영향을 주지 않았으나 NADPH oxidase 활성도와 lysosomal enzyme의 유리를 억제하였다. 식작용 중에 세포막과 세포내의 sulfhydryl기는 반응시간에 따라 점진적으로 감소하였다. N-ethylmaleiamide와 $Hg^{++}$은 세포막과 세포내의 sulfhydryl기를 모두 감소시켰다. P-Chloromercuribenzoic acid와 p-chloromercuribenzenesulfonic acid는 세포막의 sulfhydryl기를 유의하게 감소시켰으나 세포내 용해성 sulfhydryl기에는 영향을 주지않았다. Cysteine과 mercaptopropionylglycine는 superoxide 라디칼의 생성과 lysosomal enzyme의 유리를 억제하였다. Gluthathione은 superoxide생성에 영향을 주지 않았으나 뚜렷하게 lactic dehydrogenase의 유리를 억제하였다. N-ethylmaleiamide에 의한 superoxide 생성의 억제는 cysteine과 mercaptopropionyl-glycine에 의하여 반전되었으나 gluthathione의 영향은 없었다. N-ethylamleiamide에 의한 NADPH oxidase의 비활성화는 gluthathione, cysteine과 mercaptopropionylglycine에 의하여 저해되었다. Carbachol에 의하여 항진된 superoxide 라디칼 생성은 N-ethylamleiamide에 의하여 완전히 억제되었고, atropine에 의하여 길항되었다. 그러므로, 외부 자극에 대한 다형핵 백혈구 반응의 표현은 sulfhydryl기의 양의 변화와 연관이 있을 것으로 시사되었다. Lysosomal enzyme 유리는 세포막과 세포내의 sulfhydryl기에 의하여, 이에 반하여 superoxide생성은 세포내 sulfhydryl기에 의해서 영향받을 것으로 추정되었다. In opsonized zymosan activated PMN leukocytes, N-ethylamleiamide and $Hg^{++}$, penetrable sulfhydryl group inhibitors, inhibited superoxide generation, NADPH oxidase activity and lysosomal enzyme (lactic dehydrogenase and ${\beta}-glucuronidase$) secretion. P-Chloromercuribenzoic acid and p-chloromercuribenzenesulfonic acid, surface sulfhydryl group inhibitors did not affect superoxide generation but effectively inhibited both NADPH oxidase activity and lysosomal enzyme secretion. During phagocytosis, contents of surface and soluble sulfhydryl groups were gradually decreased with increasing incubation times. N-ethylmaleiamide and $Hg^{++}$ caused a loss of both surface and soluble sulfhydryl groups. P-Chloromercuribenzoic acid and p-chloromercuribenzenesulfonic acid significantly decreased the surface sulfhydryl content but did not after soluble sulfhydryl groups. Cysteine and mercaptopropionylglycine inhibited superoxide generation and lysosomal enzyme secretion. Glutathione had no effect on superoxide generation but remarkably inhibited lactic dehydrogenase release. Suppression of superoxide generation by N-ethylmaleiamide was reversed by cysteine and mercaptopropionyl-glycine but not by glutathione. Inactivation of NADPH oxidase by N-ethylmaleiamide was prevented by glutathione, cysteine or mercaptopropionylglycine. Stimulated superoxide generaion by carbachol was completely abolished by N-ethylrnaleiamide and antagonized by atropine. Thus, the expression of PMN leukocyte response to external stimuli may be associated with the change of sulfhydryl groups content. It is suggested that lysosomal enzyme secretion is influenced by both surface and soluble sulfhydryl groups, whereas superoxide generation by intracellular soluble sulfhydryl groups.

Sulfhydryl기와 세포막 구성성분의 대사 변화에 따른 다형핵 백혈구 기능의 변경

신재훈(Jeh-Hoon Shin),이정수(Chung-Soo Lee),한은숙(Eun-Sook Han),신용규(Yong-Kyoo Shin),이광수(Kwang-Soo Lee) 대한약리학회 1989 대한약리학잡지 Vol.25 No.1

면역 보체가 결합되어 있는 zymosan에 의하여 활성화된 다형핵 백혈구에서 세포 투과성 물질인 N-ethylmaleiamide과 Hg⁺⁺은 superoxide 라디칼 생성, NADPH oxidase 활성도 및 lysosomal enzyme (lactic dehydrogenase, β-glucuronidase)의 유리를 억제하였다. 세포막 단백에 특이적인 p-chloromercuribenzoic acid와 p-chloromercuribenzenesulfonic acid는 superoxide 라디칼 생성에 영향을 주지 않았으나 NADPH oxidase 활성도와 lysosomal enzyme의 유리를 억제하였다. 식작용 중에 세포막과 세포내의 sulfhydryl기는 반응시간에 따라 점진적으로 감소하였다. N-ethylmaleiamide와 Hg⁺⁺은 세포막과 세포내의 sulfhydryl기를 모두 감소시켰다. P-Chloromercuribenzoic acid와 p-chloromercuribenzenesulfonic acid는 세포막의 sulfhydryl기를 유의하게 감소시켰으나 세포내 용해성 sulfhydryl기에는 영향을 주지않았다. Cysteine과 mercaptopropionylglycine는 superoxide 라디칼의 생성과 lysosomal enzyme의 유리를 억제하였다. Gluthathione은 superoxide생성에 영향을 주지 않았으나 뚜렷하게 lactic dehydrogenase의 유리를 억제하였다. N-ethylmaleiamide에 의한 superoxide 생성의 억제는 cysteine과 mercaptopropionyl-glycine에 의하여 반전되었으나 gluthathione의 영향은 없었다. N-ethylamleiamide에 의한 NADPH oxidase의 비활성화는 gluthathione, cysteine과 mercaptopropionylglycine에 의하여 저해되었다. Carbachol에 의하여 항진된 superoxide 라디칼 생성은 N-ethylamleiamide에 의하여 완전히 억제되었고, atropine에 의하여 길항되었다. 그러므로, 외부 자극에 대한 다형핵 백혈구 반응의 표현은 sulfhydryl기의 양의 변화와 연관이 있을 것으로 시사되었다. Lysosomal enzyme 유리는 세포막과 세포내의 sulfhydryl기에 의하여, 이에 반하여 superoxide생성은 세포내 sulfhydryl기에 의해서 영향받을 것으로 추정되었다. In opsonized zymosan activated PMN leukocytes, N-ethylamleiamide and Hg⁺⁺, penetrable sulfhydryl group inhibitors, inhibited superoxide generation, NADPH oxidase activity and lysosomal enzyme (lactic dehydrogenase and β-glucuronidase) secretion. P-Chloromercuribenzoic acid and p-chloromercuribenzenesulfonic acid, surface sulfhydryl group inhibitors did not affect superoxide generation but effectively inhibited both NADPH oxidase activity and lysosomal enzyme secretion. During phagocytosis, contents of surface and soluble sulfhydryl groups were gradually decreased with increasing incubation times. N-ethylmaleiamide and Hg⁺⁺ caused a loss of both surface and soluble sulfhydryl groups. P-Chloromercuribenzoic acid and p-chloromercuribenzenesulfonic acid significantly decreased the surface sulfhydryl content but did not after soluble sulfhydryl groups. Cysteine and mercaptopropionylglycine inhibited superoxide generation and lysosomal enzyme secretion. Glutathione had no effect on superoxide generation but remarkably inhibited lactic dehydrogenase release. Suppression of superoxide generation by N-ethylmaleiamide was reversed by cysteine and mercaptopropionyl-glycine but not by glutathione. Inactivation of NADPH oxidase by N-ethylmaleiamide was prevented by glutathione, cysteine or mercaptopropionylglycine. Stimulated superoxide generaion by carbachol was completely abolished by N-ethylrnaleiamide and antagonized by atropine. Thus, the expression of PMN leukocyte response to external stimuli may be associated with the change of sulfhydryl groups content. It is suggested that lysosomal enzyme secretion is influenced by both surface and soluble sulfhydryl groups, whereas superoxide generation by intracellular soluble sulfhydryl groups.

산전MgSO₄투여와 미숙아 두개강내출혈 발생 빈도와의 관련성

황기동(Gi-Dong Hwang),신재훈(Jeh-Hoon Shin),설인준(In-Joon Seol) 대한소아신경학회 1995 대한소아신경학회지 Vol.2 No.2

연구배경: 미숙아의 두개강내 출혈빈도와 연관되는 요인들은 여러 가지가 밝혀져 있다. 임신중독증이 있는 산모에서 출생한 미숙아에서 두개강내 출혈의 발생빈도가 감소한다고 알려져 있다. 또 산전 MgSO₄투여가 미숙아 뇌출혈발생빈도를 감소시킨다는 보고가 있다. 이에 저자들은 임신중독증과 산전 MgSO₄투여에 따라 미숙아 두개강내 출혈빈도의 감소가 있는 지를 확인해보고자 본 연구를 시행하였다. 방 법: 저자들은 1991년 1월부터 1994년 12월까지 한양대학병원 미숙아실에 입원한 미숙아 174례를 대상으로 임상적 특징, 분만방법, 산모의 임신중독증 유무와 투약여부, 두 개강내출혈 유무 등을 조사하였다. 결 과: 1. 남녀비는 1:1이었고 평균 재태기간은 33.8±2.3주, 평균 출생제중은 1870 ±460gm이었다. 2. 두 개강내출혈의 발생빈도는 전체 26%이었고, 28주 미만에서는 50%, 28에서 32주의 미숙아에서는 43%, 33이상에서는 20%이었다. 3. 두 개강내출혈의 등급을 나누어 보면 GradeⅠ이 가장 많아서 33명(73%) Grade Ⅱ,Ⅲ,Ⅳ가 각각 4명 (9%), 5명(11%), 3명(7%)이었다. 4. 두 개강내출혈의 발생빈도는 복식분만에서 유의하게 낮았다.(p<0.05) 결 론: 임신중독증이 없는 산모에서 산전 MgSO₄투여에 따른 미숙아의 두개강내 출혈빈도의 감수는 없었으며, 임신중독증이 있는 산모에서 출생한 미숙아에서 두개강내 출혈빈도의 유의한 감소를 확인하였다. Background : Among the factors increasing for the intracranial hemorrhage(ICH) of premature babies. it has been reported that the presence of their maternal toxemia decreased the incidence of ICH. And also, some persons reported that the prenatal administration of MgSO₄ for their mothers decreases the incidence of ICH in premature babies. And so. we investigated retrospectively their relationship between their maternal toxemia, the prenatal administration of MgSO₄ and the incidence of ICH. Method : We investigated the 174 premature babies who were admitted to NICU of Hanyang University Hospital from Jan. 1991 to Dec. 1994. We evaluated their clinical characteristics, delivery type. medication and toxemia of their mothers, and the presence of intracranial hemorrhage(ICH). Results : 1. The mean intrauterine period was 33.8±2.3 weeks and the mean birth weight was 1870±460gm. 2. The incidence of ICH was 26% in total. 50% in below 28 weeks. 43% in 28 to 32 weeks and 20% in above 32 weeks of gestational age. 3. Grade I ICH by Papile's classification was most common. 4. When compared to vaginal delivery, there were significant decrease in ICH incidence in the cesarean section group(p<.05). 5. There was no relationship between their mother's MgSO₄ infusion and ICH incidence in groups without toxemia. 6. The incidence of ICH whose mother had toxemia was significantly lower than babies from the non-toxemic mother. Conclusion : Among the non-toxemic mothers, the incidence of ICH had no relation to whether or not the prenatal adiministration of MgSO₄. However, the incidence of ICH in the premature babies from the toxemic mothers were decreased significantly.

과체중 소아에서 천식 발생과 Adiponectin 및 Leptin의 연관성

백혜성 ( Hey Sung Baek ),김영대 ( Young Dae Kim ),박용순 ( Young Soon Park ),신재훈 ( Jeh Hoon Shin ),오재원 ( Jae Won Oh ),이하백 ( Ha Baik Lee ) 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2010 소아알레르기 및 호흡기학회지 Vol.20 No.1

목적: 전 세계적으로 증가하고 있는 비만과 천식의 연관성에 대하여 다양한 역학 조사보고들이 있지만, 확실한 결론에 이르지 못하고 있다. 본 연구에서는 과체중 소아에서 천식과 leptin과 adiponectin이 연관성이 있는지, 그리고 폐기능의 저하에 어떤 영향을 미치는지 알아보고자 하였다. 방법: 대상자는 6-18세 소아 총 61명 중 제 1군은 과체중을 동반한 안정된 천식 환아(n=14), 제 2군은 과체중을 동반하지 않은 안정된 천식 환아(n=16), 제 3군은 천식이 없는 과체중 소아(n=16), 그리고 건강한 대조군(n=15)으로 각각 분류하였다. 혈액에서 혈청 총 IgE와 알레르겐 특이 IgE치, 말초혈액 호산구 수와 혈청 호산구 음이온단백(eosinophil cationic protein: ECP)을 각각 측정하였으며, 혈액 내 leptin, adiponectin, 지질(콜레스테롤, 중성 지방, 유리지방산, 저 밀도 및 고 밀도 지질단백 콜레스테롤), insulin-like growth factor-1 (IGF-1), insulin-like growth factor-binding protein 3 (IGF-BP3)와 코티솔, 음식 섭취 설문조사 후 혈액에서 비타민 C와 E, 베타-카로틴, 셀레늄과 아연을 각각 측정하였다. 모든 환자에서 메타콜린 흡입과 자유 운동에 의한 기관지 유발시험을 각각 시행하였다. 결과: 과체중군에서 혈중 leptin치는 높은 경향이 있는 반면, adiponectin은 낮은 경향을 보였고 leptin과 adiponectin 사이에는 유의한 역 상관관계를 나타내었다. (R=-0.479; P<0.001)천식군에서 leptin은 BMI와 유의한 연관성을 보였다. 폐기능 검사로서 노력성 폐활량(forced vital capacity: FVC)에 대한 1초간 노력성 호기량(forced expiratory volume in one second: FEV1)비율, FEV1/FVC은 천식군과 비 천식군을 포함한 과체중군에서 낮았다. 하지만 IL-6와 TNF-α, 영양소 및 다른 지표들은 과체중 천식 소아에서 유의한 차이를 볼 수 없었다. 결론: 본 연구에서 측정 가능한 과체중과 관련된 비만관련 호르몬, 염증지표와 영양소들이 천식의 직접적인 유발인자로서의 역할에 대한 근거는 충분하지 않았으나 과체중이 FEV1/FVC감소 및 폐기능 저하와 연관성이 있는 것으로 사료된다. Purpose: The prevalence of asthma and obesity is increasing concomitantly, but the link between asthma and obesity is unclear. We sought to address possible roles of leptin and adiponectin in the development of asthma, and changes in pulmonary function in overweight children. Methods: Four study groups of 61 children aged 6 to 18 years (mean age, 9.69±2.16) were enrolled: (1) 14 mild-to-moderate asthmatics with overweight, (2) 16 mild-to-moderate asthmatics with normal weight, (3) 16 obese subjects without asthma, and (4) 15 healthy controls. We measured biomarkers in blood, including total and allergen-specific IgE, eosinophil, eosinophilc cationic protein (ECP), leptin, adiponectin, interleukin (IL)-6, tumor necrosis factor-α (TNF-α), lipid profiles, insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein 3 (IGF-BP3). Body mass index (BMI), antioxidants and micronutrients in a daily diet were evaluated by the questionnaire. We performed the bronchial challenge test by methacholine inhalation and free running, respectively. Results: The leptin levels was apparently high, and the adiponectin level was low in the overweight children, as depicting a significant inverse correlation between the 2 variables (R=-0.479; P<0.001). The FEV1/FVC ratio was low in the overweight children regardless of the presence of asthma. However, the effect of IL-6, TNF-α, nutrients, and other variables on asthma development in the overweight children with asthma was not verified. Conclusion: In this study, the levels of leptin, adiponectin or other obesity-related biomarkers were not independently associated with asthma. Therefore, it is concluded that obesity may not be an important factor in pulmonary function impairment. [Pediatr Allergy Respir Dis(Korea) 2010;20:48-58]

선천성 부신 과형성

신재훈 한국의학사 1989 診斷과治療 Vol.9 No.5

성장지연의 진단 및 치료

신재훈 한국의학사 1988 診斷과治療 Vol.8 No.10

선천성 갑상선종성 기능저하증

신재훈 한국의학사 1989 診斷과治療 Vol.9 No.8

열량제한 백서에서 1 형 인슐린양 성장인자의 효과 : 1 형 인슐린양 성장인자 투여시 혈중농도를 정상으로 회복하나 장기 저항에 관한 연구 1 injection to promote growth in energy restricted rats despite normalization of serum IGF - 1 concentration

신재훈 대한내분비학회 1994 Endocrinology and metabolism Vol.9 No.3

In addition to regulation mediated by CH, nutritional status is an important regulator of IGF-Ⅰ, circulating levels of these peptides modulated by poor nutritional status and poorly controlled insulin dependent diabetes mellitus(IDDM). Impaired growth is a well recognized complication of poorly controlled diabetes. This study was designed to examine the anabolic effect of recombinant human IGF-I in energy-restriction model. Experimental design; Sprague-Dawley rats(n=20) weighing 90-100g were used. Rats were fed a control diet two times a day(AM 8-11, PM 5-8) for four days after arrival and then assigned to one of three groups: control, energy-restricted, energy-restricted IGF-I treatment group. Energy restricted group was given with a decrese of 25% in the energy without changes in the protein by feeding 88% by weight to energy-restricted diet. During the 10days of energy restriction, the growth rate was reduced by 35%(2.70+-0.18g/day in energy restricted group vs. 4.13+-0.75g/day in the control group). At sacrifice, the tail lengh and weight of organs were not significantly decreased except the spleen and thymus(-17%: P$lt;0.05). Serum IGF-I was reduced by 19% at the end of 10days of energy restriction. The glycemia, measured each day by glucometer from blood collected at the tail, was not reduced by energy restriction(105.4+-7.7 in control group vs. 101.3+-4.1㎎/㎗). The abundance of serum IGF-BPs was unchanged by this restriction. Despite the 1.5 fold increase of IGF-I concentration in energy restricted IGF-I injection group at sacrifice(1994+-172ng/㎖ vs. 1221+-110 ng/㎖ energy restricted group), IGF-I treatment(300 ㎍/day in twice sc injection for 6day) did not significantly accelerate the growth rate(body weight)(2.87+-0.20 vs. 2.70+-0.18g/day in energy restricted group). The glycemia was slightly reduced by IGF-I treatment(91.7+-5.0 ㎎/㎗ vs. 101.3+-4.5 ㎎/㎗ in energy restricted group), but it was not significant. However, the spleen and thymus weight, decreased by energy restriction, was completely normalized by IGF-I treatment. In summary, lack of a significant anabolic response to injection of IGF-I during energy restriction in this study may be associated with the compensatory growth response(alterations in dietary protein utilization) which followed initial period of energy restriction(J Kor Soc Endocrinol 9: 213-218, 1994).

고압 액상 크로마토그래피를 이용한 혈중 성장홀몬-결합단백 측정에 대한 초기보고

신재훈 대한내분비학회 1993 Endocrinology and metabolism Vol.8 No.4

A technique high pressure liquid chromatography (HPLC) gel filtration was used to evaluate GH-binding proteins in human serum. Seurm (75 l) was well seperated from free Biotin-hGH(PeakIII). Elution profile was monitored in potical density multisaner system. In chromatography, the molecular weight of Peak I and II appeared about 200 K and 60 K dalton respectively. In control serum, the specific binding of biotin-HGH to Peak II-BP was 59.7+/-0.8% of the optical density. Specific binding of Biotin hGH to Peak Ii-BP was low during the infant period and progressively increase as age. Using gel filltration system, binding of biotin-hGH by normal adult serum was dependent on time (maximum equlibrium was reached in 24h at 4C), temperature( 4C>22C>37C)and seurm concentration. These data indicated that human sera contain a specific binding protein for hGH. The biological significance of this binding protein remains to be investigated. We sugest the measurement of GH-BP II activity by HPLC provides a clear seperation. Our nonradioactive optical detection method by multiscaner was also highly resolution (J Kor Soc Endocrinol 8: 432438, 1993)

임상영양지표 ( Nutritional Index )로서의 1형 인슐린양 성장인자 ( IGF-I )의 역할

신재훈 대한내분비학회 1994 Endocrinology and metabolism Vol.9 No.2