토끼 십이지장구의 운동성에 미치는 dopamine의 영향

이윤렬,신원임,박형진,Lee, Yun-Lyul,Shin, Won-Im,Park, Hyoung-Jin 대한생리학회 1986 대한생리학회지 Vol.20 No.2

dopamine이 십이지장구를 포함한 소장의 운동성에 미치는 영향, cholecystokinin이 dopamine의 작용에 미치는 영향 그리고 이들 작용의 신경성 기전을 알아보고자 다음과 같은 실험을 실시하였다. ether로 마취한 토끼 54마리에서 십이지장구, 십이지장, 공장 그리고 회장을 적출하고 절편(길이 1cm)을 만들어 Krebs-Ringr 용액이 채워진 기록 용기에 넣고 자발적인 등장성 수축을 기록하였다. Krebs-Ringr 용액에는 5% $Co_2$를 함유하는 $O_2$를 계속 공급하였으며, 용액의 온도가 $37^{\circ}C$를 유지하도록 하였다. 자발적 수축이 시작하고 20분이 경과한 다음 dopamine($10^{-4}M$), CCK-8($10^{-8}M$), dopamine($10^{^6}M$)등을 투여하면서 수축성을 관찰하여 다음과 같은 결과를 얻었다. 1) dopamine은 소장의 모든 부위에서 자발적 수축성을 억제하였으며, 이러한 dopamine의 작용은 회장을 제외한 다른 부위에서 tetrodotoxin에 의하여 유의하게 감소하였다. 2) domperidone은 소장의 모든 부위에서 dopamine의 억제작용에 길항적으로 작용하였으며, tetrodotoxin을 전처치하면 회장을 제외한 다른 부위에서 domperidone의 길항작용은 완전히 소실되었다. 3) CCK-8는 소장의 모든 부위에서 dopamine의 작용을 감소시켰으며, tetrodotoxin을 전처치하면 CCK-8의 작용은 유의하게 감소되었다. 이상의 결과로 미루어 보아 dopamine은 십이지장구를 포함한 모든 소장의 수축성을 억제하며, CCK-8는 dopamine의 억제 작용을 감소시키는데, 이러한 작용들은 장관내 신경계를 거쳐서 간접적으로 일어나는 것으로 생각된다. The present study was undertaken to see an interaction of dopamine and cholecystokinin on spontaneous contractility of the small intestine including the duodenal bulb. A possible neural mechanism of the interaction was alto examined. The spontaneous isometric contractility of a segment of the duodenal bulb, duodenum, jejunum and ileum obtained from the rabbit anesthetized with ether was recorded in a chamber filled with Krebs-Ringer's solution. The solution was constantly kept at $37^{\circ}C$ and aerated with $O_2$ containing 5% $CO_2$. After 20 min from beginning of the contraction, dopamine $(10^{-4}M)$, CCK-8($10^{-8}M$), domperidone($10^{-5}M$) and tetrodotoxin ($10^{-6}M$) were administered into the chamber The following results were obtained by analyzing changes in the contractility of the intestinal segments. 1) Dopamine inhibited the spontaneous contractility of the duodenal bulb, duodenum, jejunum and ileum. The inhibitory action of dopamine on all parts of the small intestine except the ileum was reduced by tetrodotoxin. 2) Domperidone knwon to be a specific peripheral dopamine receptor antagonist blocked the inhibitory action of dopamine on all parts of the small intestine. The antagonistic action of domperidone on all parts of the small intestine except the ileum was completely abolished by tetrodotoxin. 3) CCK-8 reduced the inhibitory action of dopamine on all parts of the small intestine. The effect of CCK-8 on the dopamine action was diminished by tetrodotoxin. These results suggest that dopamine inhibits the spontaneous contractility of the small intestine including the duodenal bulb and CCK-8 reduces the inhibitory action of dopamine through the enteric nervous system.

송아지 설사에 영향을 미치는 사육밀도 및 축사 방위와의 상관관계에 관한 연구

이윤렬 ( Yun Lyul Lee ),이학림 ( Hak Rim Lee ),안재범 ( Jae Bum Ahn ),송지예 ( Ji Ye Song ),장재진 ( Jae Jin Jang ),이민재 ( Min Jae Lee ) 한국동물위생학회 2007 韓國家畜衛生學會誌 Vol.30 No.1

This study was conducted to determine the effective environmental control measures in preventing calf diarrheal disease in farms. It was found that the high numbers of calves in a cow house was directly related with the high incidence rate of diarrheal diseases. When the number of calves was increased in a cow house, it was difficult to reduce the diarrheal diseases of calves. It was revealed that the incidence rates of calf diarrheal disease were dramatically reduced when the delivery house which faced south. The periodical changes of bedding materials seemed beneficial good to control the possible resident infectious agents, including pathogenic viruses and bacteria. The control of the delivery time was important for reduction of calf diarrhea. It was revealed that September and October were the best time as the delivery period. In order to use old cow house as a delivery house, it should be sterilized by the disinfectant and the flame. When the diarrheal feces were removed as soon as possible, the transmission of diarrheal diseases among calves in the same house was reduced. When the diarrheal feces were not disposed, the incidence rate of diarrhea reached up to 50%. When the morbid calves were not quarantined, the incidence rate of diarrhea was 100%. In contrast, when the diseased calves were quarantined from other calves, the incidence rate of diarrhea was reduced, remarkably. The mean recovery time from the diarrhea of the treated calves in the southern cow house was much faster than that in the north cow house. The treatment of calves in a clean and well-ventilated cow house with dry bedding was more effective than that in a cold, wet, and dark house. It could be postulated that the most important environmental factor in the reduction of diarrheal diseases of calves is the blocking of the entry of the infectious agents. The complete sanitation and control of the delivery house in the using farm is more important than building a new cow house, for prevention of calf diarrheal diseases.

강원도 지역에 서식하는 무당개구리에서 생물학적 활성을 지닌 Bombesin 유사 Peptide 에 관한 연구

박형진,이윤렬,권혁일,김일,유일재,D . R . Marshak ( Hyoung Jin Park,Yun Lyul Lee,Hyeok Yil Kwon,Yil Kim,Il Je Yu,Daniel r . Marshak ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.3

The present study was performed to purify a BLI from the skin of frogs, B. orientalis inhabiting Kangwon-Do, Korea, and then determine its physiological activities and amino acid sequence. A substance that shows bombesin-like immunoreactivity (BLI) was partially purified from crude methanol extract of the frog skin by using a column of alkaline alumina and Sephadex G-10. BLI was further purified by using RP C18 preparative HPLC in which BLI was separated into 5 peaks. The major peak (BLI-K1) containing 65% of total BLI was subjected to purify as a single component by using sequential HPLC of SP-ion exchange and RP C18. Content of BLI in each fraction was monitored by RIA for which bombesin antiserum with a high titer and affinity to BBS was raised in a guinea pig. Eventually, 890 ㎍ of BLI-K1 was homogeneously purified from 420g of the skin of B. orientalis and it was not differentiated from synthetic BBS in RP C18, gel permeation and SP-ion exchange HPLC. In the physiological activities, BLI-K1 was identical to synthetic BBS in stimulation of gastrin release and exocrine pancreatic secretion including volume, protein, bicarbonate, amylase and chymotrypsin output in anesthetized rats. Furthermore, molecular weight and partial amino acid sequence of BLI-K1 was revealed to be identical to synthetic BBS. Therefore, it is concluded from the above results that the skin of B. orientalis contains the same BBS that has been isolated from the skin of European Bombina.

강원도 지역에 서식하는 무당개구리에서 생물학적 활성을 지닌 Bombesin 유사 Peptide에 관한 연구

박형진,이윤렬,권혁일,김일,유일재,Park, Hyoung-Jin,Lee, Yun-Lyul,Kwon, Hyeok-Yil,Kim, Yil,Yu, Il-Je 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.3

본 실험에서는 강원도 지역에 집단으로 서식하고 있는 한국산 무당개구리(Bombina orientalis) 피부에서 bombesin 유사 peptide를 순수정제하여 생리활성과 amino acid 서열을 측정하고자 하였다. 무당개구리의 피부를 methanol에 담가 bombesin 유사반응물질(BLI)을 추출한 다음 alkaline alumina에 이은 Sephadex G-10 겔 크로마토그래피와 RP C18 preparative HPLC로 부분정제하였다. 부분정제시 여러 종류의 BLI가 출현하므로 본 연구에서는 전체 BLI의 65%를 점유하는 peak(BLI-K1)를 RP C18 및 SP-이온교환 HPLC를 이용하여 순수정제하였다. 피부조직 420 g에서 $890\;{\mu}g$의 BLI-K1를 순수정제하였으며, 이것은 RP C18, 겔 투과 그리고 SP-이온교환 HPLC 모두에서 합성 bombesin(BBS)과 동일한 정체시간을 나타내었다. 생리활성에서도 BLI-K1은 마취한 흰쥐에서 혈장 gastrin 농도와 혜장 단백질, 중탄산이온, amylase 및 chymotrypsin 분비량을 모두 유의하게 증가시켜 합성 BBS와 동일한 활성을 나타내었다. 또한 BLI-K1의 분자량 및 C-terminal의 11개 amino acid 서열이 합성 BBS과 동일하였기 때문에 한국산 무당개구리의 피부에도 유럽산 무당개구리의 피부에 존재하는 것과 동일한 bombesin이 존재하는 것으로 생각된다. The present study was performed to purify a BLI from the skin of frogs, B. orientalis inhabiting Kangwon-Do, Korea, and then determine its physiological activities and amino acid sequence. A substance that shows bombesin-like immunoreactivity (BLI) was partially purified from crude methanol extract of the frog skin by using a column of alkaline alumina and Sephadex G-10. BLI was further purified by using RP C18 preparative HPLC in which BLI was separated into 5 peaks. The major peak (BLI-K1) containing 65% of total BLI was subjected to purify as a single component by using sequential HPLC of SP-ion exchange and RP C18. Content of BLI in each fraction was monitored by RIA for which bombesin antiserum with a high titer and affinity to BBS was raised in a guinea pig. Eventually, $890\;{\mu}g$ of BLI-K1 was homogeneously purified from 420g of the skin of B. orientalis and it was not differentiated from synthetic BBS in RP C18, gel permeation and SP-ion exchange HPLC. In the physiological activities, BLI-K1 was identical to synthetic BBS in stimulation of gastrin release and exocrine pancreatic secretion including volume, protein, bicarbonate, amylase and chymotrypsin output in anesthetized rats. Furthermore, molecular weight and partial amino acid sequence of BLI-K1 was revealed to be identical to synthetic BBS. Therefore, it is concluded from the above results that the skin of B. orientalis contains the same BBS that has been isolated from the skin of European Bombina.

적출한 고양이 위(胃)의 전기활동에 미치는 prostaglandin $E_2$ 및 indomethacin의 영향

김명석,이윤렬,Kim, Myung-Suk,Lee, Yoon-Lyur 대한생리학회 1983 대한생리학회지 Vol.17 No.1

This study was conducted to investigate the effect of prostaglandin $E_2(PGE_2)$ and indomethacin upon the electrical activity of the isolated cat stomach muscle strips$(1.5{\times}7.0\;cm)$. Fifty-seven muscle strips, obtained from 57 cat stomachs(including corpus and antrum) were studied in a muscle chamber filled with Krebs solution(pH 7.4, temperature $36{\pm}0.5^{\circ}C$) aerated with 5% $CO_2$ in $O_2$. The electrical activity was recorded by five capillary electrodes (Ag-AgCl), of which two were placed on the corpus and three on the antrum. After recording of the electrical activity in normal Krebs solution, $PGE_2$ in concentrations of 0.25(N=7), 0.5(=7), 1(N=7) and $2{\times}10^{-7}\;M(N=6)$ were administered to 27 muscle strips, while indomethacin was applied in concentretions of 0.25(N=9), 0.5(N=10), 1(N=6) and $2{\times}10^{-3}\;M(N=5)$ to the remaining 30 strips. The mean frequency were minutely measured from each electrogastrogram. 1) By adding $PGE_2$ in all doses, gastric slow wave frequency increased significantly compared with that in resting state. 2) Following $PGE_2$ administration, peak slow wave frequency increased dose-dependently. 3) After indomethacin addition in all doses, the slow wave frequency decreased significantly compared with that in resting state. 4) Following indomethacin administration, incidence of complete abolition of slow wave increased dose-dependently, and its latent period decreased also in a dose-dependent manner. It is inferred from the above results that prostaglandin $E_2$ has a facilitatory role in the development of gastric slow wave in cat.

Effect of Reserpine on Pancreatic Exocrine Secretion Induced by Mesencephalic Reticular Stimulation in Rats

박형진,이윤렬,Park, Hyoung-Jin,Lee, Yun-Lyul The Korean Physiological Society 1988 대한생리학회지 Vol.22 No.1

최근에 마취한 흰쥐에서 중뇌망상체를 전기적으로 자극하면 췌장의 외분비 기능이 증가하며 이러한 결과는 망상체의 자극으로 인하여 교감신경계의 활성도가 상승하기 때문이라는 보고가 있다. 한편 교감신경계의 활성도가 상승할 경우 교감신경계의 전달 물질인 catecholamine이 교감신경 종말 뿐만 아니라 부신수질에서도 유리된다고 알려져 있다. 그러므로 본 연구에서는 중뇌망상체의 자극으로 인하여 췌장의 외분비 기능이 증가함에 있어 교감신경계가 중요한 역할을 담당하는지를 확인하고, 이때 부신수질이 관여하는가를 알아보고자 하였다. 마취한 흰쥐에게 atropine (1mg/kg) 또는 reserpine (5mg/kg)을 투여하거나 또는 부신을 적출한 다음 중뇌망상체를 전기 자극하면서 췌장액을 채취하였다. 사용한 전기자극의 매개변수는 1.3V, 40Hz, 2msec이었다. atropine과 reserpine을 투여하면 마취한 흰쥐의 자발적 췌장액 분비량과 단백질 분비량은 모두 유의하게 감소하였으나 부신을 제거하면 췌장액 분비량에는 이렇다할 변동이 없는 반면에 단백질 분비량은 유의하게 감소하였다. 중뇌망상체를 전기자극하면 췌장액 분비량과 단백질 분비량 모두가 유의하게 증가하였다. 이러한 망상체의 자극효과는 atropine 전처치에 의하여 이렇다할 영향을 받지 않았으나 reserpine 전처치에 의하여 소실되었다. 그러나 부신을 적출하면 망상체 자극에 의한 췌장액 분비량의 증가는 유지되는 반면에 단백질 분비량의 증가는 소실되었다. 한편 미주신경을 절단한 흰쥐에서 중뇌망상체를 자극하는 동안에 경동맥의 수축기 및 이완기 혈압이 상승하였는데 이러한 망상체의 자극효과도 reserpine의 투여에 의하여 유의하게 감소되었다. 본 실험의 결과를 종합하여 보면 마취한 흰쥐에서 중뇌망상체의 자극은 교감신경계를 활성화시켜 췌장액 분비량과 단백질 분비량에 촉진적인 영향을 미치며, 이때 활성화된 교감신경계는 부분적으로 부신을 경유하게 췌장의 단백질 분비에 촉진적인 영향을 미치는 것으로 생각된다.

깨죽이 정상 성인의 혈장 Gastrin 농도에 미치는 영향

김명석,이윤렬,권경옥,조양혁,박형진,Kim, Myung-Suk,Lee, Yoon-Lyeur,Kwon, Kyoung-Ok,Jo, Yang-Hyeok,Park, Hyoung-Jin 대한생리학회 1982 대한생리학회지 Vol.16 No.2

This study was conducted to investigate the effect of ingestion of sesame (Sesamum indicum) gruel as a nourishing meal upon the plasma gastrin concentration in normal Korean. Sixteen normal persons with no history of gastrointestinal diseases, including male and female were studied. After an overnight(about 15 hrs) fast, eight persons(mean age: 26.6, range: $20{\sim}40$ years) of them ingested a 350 ml sesame gruel corresponding to 12 g protein, 13 g fat and 99 g carbohydrate, and the remaining 8 subjects(mean age: 21.3, range: $20{\sim}24$ years) ingested a 350 ml glutinous rice gruel(control meal) corresponding to 8 g protein, 1 g fat and 115 g carbohydrate. The venous blood samples were drawn before and after the ingestion of the test meal for the measurement of gastrin by means of radioimmunoassay. 1) Plasma gastrin concentration in response to the ingestion of sesame gruel or glutinous rice gruel increased significantly compared with the concentration in fasting state. 2) Mean increment or percent increment in postprandial plasma gastrin concentration after the ingestion of sesame gruel was not significantly different from that after the control meal, i.e. the glutinous rice gruel. It is inferred from the above results that the ingestion of sesame contained in sesame gruel may have no significant influence on gastrin release in normal human subjects.

방사면역측정을 위한 항 Gastrin 혈청의 생산 및 평가

박형진,권혁일,이윤렬,신원임,서상원,오양석,Park, Hyoung-Jin,Kwon, Hyeok-Yil,Lee, Yun-Lyul,Shin, Won-Im,Suh, Sang-Won,Oh, Yang-Suk 대한생리학회 1989 대한생리학회지 Vol.23 No.1

New Zealand white 계의 토끼에게 EDC를 사용하여 hemocyanin에 접합시킨 human G-17-I을 면역한 결과 gastrin의 방사면역측정에 적합한 항 gastrin 혈청을 얻는데 성공하였다. 본 실험에서 얻은 항 gastrin 혈청의 역가는 1:25,000이며 human G-17-I과 human big gastrin에 매우 친화력이 높은 반면 다른 위장관 peptide 들과는 교차반응성이 아주 낮았다. 본 항 gastrin 혈청의 effective affinity constant는 $1.77{\times}10^{11}/M$이었다. 또한 총 결합부위(total binding sites)는 255 nM, 이질성지수(heterogeneity index)는 0.84 그리고 average affinity constant는 $0.79{\times}10^{11}/M$이었다. 본 실험에서 얻은 항 gastrin 혈청을 사용하여 사람과 흰쥐의 혈장 gastrin 농도를 방사면역측정법으로 측정한 결과 이미 잘 알려진 결과와 동일한 성적을 얻을 수 있었다. 이러한 결과들을 종합하여 볼때 본 실험에서 얻은 항 gastrin 혈청은 생리적농도로 존재하는 gastrin의 방사면역측정에 적합한 것으로 생각된다. In order to produce antibody for use in radioimmunoassay of gastrin in physiological concentration, four rabbits of New Zealand white were immunized with synthetic human gastrin-17-I conjugated to hemocyanin with EDC. Among them, only one rabbit produced antibody that could bind 50% of $^{125}I-gastrin$ at a final dilution of 1:25,000. $^{125}I-gastrin$ was prepared with synthetic human gastrin-17-I and $NaI^{125}$ by lactoperoxidase technique. The product was then purified on a column of Sephadex Gl5/G5O (7:3, w/w) followed by a column of DEAE sephadex A-25. The specific radioactivity of the purified $^{125}I-gastrin$ was in the range of 347-1429 ${\mu}Ci/nmole$ when determined by the self-displacement method. The effective affinity constant $(K_{eff})$, total binding sites (N), heterogeneity index $({\alpha})$ and average affinity constant $(K_{0})$ of the anti-gastrin serum calculated from Scatchard plot as well as Sips plot were $1.77{\times}10^{11}/M$, 255 nM, 0.84 and $0.79{\times}10^{11}/M$, respectively. When radioimmunoassay was performed with the anti-gastrin serum, it was confirmed that the mean concentration of gastrin immunoreactivity in plasma was increased by feeding in humans and rats, and also increased by bombesin administration in rats. The results indicate that the anti-gastrin serum produced in the present investigation is suitable for radioimmunological determination of gastrin in physiological concentration.

적출한 고양이 위(胃)의 전기활동에 미치는 prostaglandin E₂ 및 indomethacin의 영향

김명석(Kim, Myung-Suk),이윤렬(Lee, Yoon-Lyur) 대한생리학회 1983 대한생리학회지 Vol.17 No.1

This study was conducted to investigate the effect of prostaglandin E₂(PGE₂) and indomethacin upon the electrical activity of the isolated cat stomach muscle strips(1.5 X 7.0 cm). Fifty-seven muscle strips, obtained from 57 cat stomachs(including corpus and antrum) were studied in a muscle chamber filled with Krebs solution(pH 7.4, temperature 36±0.5℃) aerated with 5% CO₂ in O₂. The electrical activity was recorded by five capillary electrodes (Ag-AgCl), of which two were placed on the corpus and three on the antrum. After recording of the electrical activity in normal Krebs solution, PGE₂ in concentrations of 0.25(N=7), 0.5(=7), 1(N=7) and 2 X 10^-7 M(N=6) were administered to 27 muscle strips, while indomethacin was applied in concentretions of 0.25(N=9), 0.5(N=10), 1(N=6) and 2 X 10^-3 M(N=5) to the remaining 30 strips. The mean frequency were minutely measured from each electrogastrogram. 1) By adding PGE₂ in all doses, gastric slow wave frequency increased significantly compared with that in resting state. 2) Following PGE₂ administration, peak slow wave frequency increased dose-dependently. 3) After indomethacin addition in all doses, the slow wave frequency decreased significantly compared with that in resting state. 4) Following indomethacin administration, incidence of complete abolition of slow wave increased dose-dependently, and its latent period decreased also in a dose-dependent manner. It is inferred from the above results that prostaglandin E₂ has a facilitatory role in the development of gastric slow wave in cat.

ρ-Chlorophenylalanine이 랫드 췌장의 외분비기능에 미치는 영향

박형진(Park Hyoung-Jin),이윤렬(Lee Yun-Lyul),서상원(Suh Sang-Won),김길수(Kim Kil-Soo),윤효섭(Yoon Hyo-Seop),허수정(Heo Su-Jeong),윤석주(Yoon Seok-Joo) 한국실험동물학회 1992 Laboratory Animal Research Vol.8 No.1