왕겨에 의한 신령버섯균사체 액체배양액의 생쥐 항복수암성 증가

Young S. Kim(김영숙),Wook J. Jang(장욱진),Md. A. Rakib(라키브),Jung M. Kwon(권정민),Chae R. Ahn(안채린),So Y. Kim(김소영),Yong U. Cho(조용운),Young K. Ha(하영권),Jeong O. Kim(김정옥),Yeong L. Ha(하영래) 한국생명과학회 2010 생명과학회지 Vol.20 No.9

왕겨가 Agaricus blazei Murill (AB: 신령버섯)균사체 액체배양 추출물의 항암성을 증가시키는지에 관한 연구를 수행하였다. AB균사체를 대두박을 기본으로 한 액체배지에 다양한 조건으로 배양하여, β-glucan 함량, Brix, 균사체를 측정하여 적정 생육조건을 선정하고, 이들의 열수추출물의 S-180 cell로 유도한 mouse 복수암에 대한 항암성을 조사하였다. AB균사체는 25℃에서 5일간 배양하였을 때 최적 생육을 나타내었고, 이 배양물이 다른 조건에서 배양한 배양물보다 우수한 항암성을 나타내었다. AB균사체의 생육 및 항암성은 변온배양에 따른 효과는 없었다. 따라서 이 최적배양조건(25℃, 5일 배양)에서 AB균사체를 1% 왕겨분말이 함유된 액체배지에 배양하고, 이의 열수 추출물의 항암성을 검증하였다. 1% 왕겨가 함유된 액체배지에서 배양한 열수추출물의 항암성은 왕겨가 함유되지 않은 배지의 열수추출물보다 항암성이 유의성 있게 증가되었다(p<0.05). 왕겨의 첨가는 AB균사체의 생육을 오히려 촉진시켰다. 이 결과는 왕겨가 AB균사체 뿐 만 아니라 다른 버섯균사체 액체배양물의 항암성 증진을 위한 원료로 활용될 수 있을 것임을 의미한다. The effects of rice hull (RH) powder on the anticarcinogenic activity of submerged-liquid cultures of Agaricus blazei Murill (AB) were assessed for mouse ascites cancers induced by mouse Sarcoma S-180 (S-180) cancer cells. Optimal growth of AB mycelia in the basal liquid culture medium, containing soybean meal, was achieved by culturing at 25℃ for 5 days, when evaluated by β-glucan content, Brix, and mycelial weight, relative to other culture conditions. Hot-water extract (HWE) of the submergedliquid culture of AB mycelia grown at 25℃ for 5 days exhibited a stronger anticarcinogenic activity, relative to HWE from other culture conditions. No such effects were obtained from AB mycelial cultures by alternative temperature-controlling cultures. Both cytotoxicity for S-180 cells and anticarcinogenic potentials for mouse ascites cancer of the HWE from AB mycelia grown in the basal medium containing 1% RH powder for 5 days at 25℃ were significantly (p<0.05) enhanced, relative to HWE from the AB mycelia culture of the basal medium without RH powder. These results indicate that HWE of submerged-liquid culture of AB mycelia, incubated in media containing 1% RH powder at 25℃ for 5 days, enhanced anticarcinogenic activity against S-180 cell-induced mouse ascites cancer, and suggest that RH powder is an excellent ingredient for the improvement of the anticarcinogenic potentials of the submerged-liquid culture of mushroom mycelia.

Optimal Level for the Protection of Carbon Tetrachloride-induced Sprague-Dawley Rat Liver Damage by Mycelial Cultures of Lentinus edodes

Wook J. Jang(장욱진),Young S. Kim(김영숙),Yeong L. Ha(하영래),Cherl W. Park(박철우),Young K, Ha(하영권),Jeong O. Kim(김정옥) 한국생명과학회 2010 생명과학회지 Vol.20 No.5

표고버섯균사체 고체배양물 분말과 액체배양물 분말을 2:1 (w/w)로 혼합한 시료 (LED로 명명)가 CCl₄로 유발된 숫컷 쥐의 간독성을 보호하는 효과에 대해 연구하였다. Vehicle 처리군(0.2 ㎖ 증류수), Control 처리군(0.2 ㎖ 증류수), LED 처리군(LED 100, 200, 300, 400 ㎎/㎏ BW을 0.2 ㎖ 증류수에 각각 혼합), 그리고 Silymarin 처리군(200 ㎎/㎏을 0.2 ㎖ 증류수에 용해)의 각 처리군(군당 여섯 마리)에 매일 2주간 투여한 다음, Vehicle 처리군을 제외한 모든 군에 CCl₄ (CCl₄:corn oil, 1:1 v/v; 0.5 ㎖/㎏ BW)를 복부에 주사하였다. 하루가 지난 후에 생화학적 지표성분을 분석하기 위해 혈액과 간 시료를 채취하였다. 모든 LED 처리에 의해 간 SOD, catalase, 그리고 GSH peroxidase의 활성이 증가하였으며, TBARS, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) 그리고 interleukin-6 (IL-6)가 감소하였다. 그 결과, 혈청중의 GOT, GPT, 그리고 LDH의 활성이 감소하였다. LED의 간 기능보호효과가 가장 효과적으로 나타난 처리농도는 200 ㎎/㎏ BW였다. 이러한 결과로 보아 LED는 CCl₄로 유발된 SD 쥐의 간 손상을 보호하며, 이는 LED의 항산화능과 cytokine의 감소에 의한 것이다. 따라서, LED는 사람의 간독성보호를 위해 효과적으로 사용할 수 있을 것으로 생각된다. The protective effects of a powder mixed with solid-cultured and liquid-cultured Lentinus edodes mycelia (2 : 1, w/w) (designate LED) with different doses of carbon tetrachloride (CCl₄) on induced hepatotoxicity in male Sprague-Dawley (SD) rats was investigated. The rats were divided into seven groups (6 rats/group) and the following substances were administered orally to each group: Vehicle (0.2 ㎖ distilled water), Control (0.2 ㎖ distilled water), LED (LED 100, 200, 300 and 400 ㎎/㎏ BW in 0.2 ㎖ distilled water), and Silymarin (200 ㎎/Kg BW in 0.2 ㎖ distilled water). After two weeks of daily administration, all groups except for the Vehiclegroup were subjected to abdominal injection with CCl₄ (CCl₄ : corn oil, 1 : 1 v/v; 0.5 ㎖/㎏ BW). One day later, blood and liver samples were collected to analyze biomarkers. All LED treatments elevated hepatic superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH peroxidase) activities, and reduced thiobarbituric reactive substances (TBARS), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6), resulting in the reduction of glutamate-oxalate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and lactic acid dehydrogenase (LDH) activities in plasma. These results indicate that LED effectively protected SD rat hepatotoxicity induced by CCl₄ through its antioxidative activity and reduction of some cytokines. The highest efficacy was found in LED 200 ㎎/㎏ BW, showing potential as a useful material for protection from hepatotoxicity in humans.

Mycelial Culture of Lentinus edodes Alleviates Rat Liver Toxicity Induced by Carbon Tetrachloride and Ethanol

Yeong L. Ha(하영래),Young S. Kim(김영숙),Chae R. Ahn(안채린),Jung M. Kweon(권정민),Cherl W. Park(박철우),Young K. Ha(하영권),Jeong O. Kim(김정옥) 한국생명과학회 2010 생명과학회지 Vol.20 No.1

LED의 간 보호 기능을 연구하기 위하여 CCl₄ 및 ethanol로 SD rat에 간독성을 유발한 다음, LED를 처리하였다. LED의 간 기능 보호효과는 간장치료제인 Silymarin과 비교하였다. CCl₄로 간 독성을 유발한 경우, LED는간의 항산화효소인 SOD, catalase, GSH peroxidase 효소활성의 항진을 유도하였고, 산화물인 TBARS의 함량을 감소시켰다. 또한 간 손상의 지표인 혈장의 GOT, GPT 및 LDH의 활성을 감소시켰다. Ethanol로 간 독성을 유발한 경우 LED는 간의 SOD, catalase, GSH preoxidase 효소활성 및 GSH 함량을 항진시켰고, 총 cholesterol, triglyceride 및 TBARS의 함량을 감소시켰다. 또한 ethanol 대사에 관여하는 ADH 효소 활성을 증진시켰고, ROS 생성에 관여하는 CYP2E1 효소의 발현을 감소시킴으로써, 혈장의 GOT, GPT 및 LDH 효소활성이 감소되었다. 또한 LED는 DPPH 및 mouse liver mitochondrial system에서 항산화효과를 보였다. 이러한 결과로 미루어 볼 때 LED는 in vitro와 in vivo에서 항산화효과에 의한 간 기능 보호효과를 갖는 것으로 추정된다. The protective effect of a mixed powder from solid-cultured and liquid-cultured Lentinus edodes mycelia (2:1, w/w) (designate LED) on the carbon tetrachloride (CCl₄)- and ethanol-induced hepatotoxicity of male Sprague-Dawley (SD) rat was investigated. In the CCl₄-induced rat hepatotoxicity experiment, rats of 4 groups (6 rats/group) were administere with Normal (0.2 ml distilled water), Control (0.2 ml distilled water), LED (LED 200 ㎎/㎏ BW + 0.2 ml distilled water), and Silymarin (200 ㎎/㎏ BW + 0.2 ml distilled water), p.o., daily for 2 weeks. Afterwards, all groups except for the Normal group were subjected to abdominal injection with CCl₄ (CCl₄: corn oil, 1:1 v/v; 0.5 ml/㎏ BW). For the ethanol- induced rat hepatotoxicity experiment, rats were divided into 5 groups (5 rats/group): Normal; Pair-fed control (PFC); Control (ethanol); LED (ethanol + LED 200 ㎎/㎏ BW); and Silymarin (ethanol + silymarin 200 ㎎/㎏ BW). Rats of the Normal and PFC groups were fed a basal liquid diet, and rats of the Control, LED, and Silymarin groups were fed a liquid ethanol diet containing LED or Silymarin. Eight weeks later, blood and liver samples were collected to analyze biomarkers. In CCl₄-induced SD rats, LED elevated hepatic superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH peroxidase) activities and thiobarbituric reactive substances (TBARS) were reduced, resulting in the reduction of glutamate-oxalate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and lactic dehydrogenase (LDH) activities in plasma. Similar results of these enzymes and biochemical markers in both liver tissues and plasma were seen in ethanol-induced hepatotoxicity of SD rats. In addition, elevated alcohol dehydrogenase (ADH) activity and reduced expression of cytochrome p450 mixed monooxygenase enzyme (CYP2E1) were seen in liver tissues from ethanol-treated rats by LED treatment. These effects of LED were similar to those of Silymarin. In in vitro experiments, LED showed antioxidant activity in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) system and mouse liver mitochondria system induced by NADPH/Fe<SUP>2+</SUP> and cumine hydroperoxide (CuOOH). These results indicate that LED protected SD rat hepatotoxicity, induced by CCl₄ and ethanol, through its antioxidative activity and might be useful as a material for protection from hepatoxicity in humans.

CLA의 생물학적 기능

하영래(Yeong L. Ha),김정옥(Jeong O. Kim),김영숙(Young S. Kim) 한국생명과학회 2017 생명과학회지 Vol.27 No.8

CLA는 탄소가 18개(C18)인 linoleic acid의 이중결합이 C9,C11; C10,C12 위치로 이동됨으로 생성되는 입체이성(c,c; c,t; t,c; t,t)체를 총괄적으로 일컷는 말이다. 이론적으로 가능한 이성체 중에서 c9,t11-CLA가 rumen bacteria, lactic bacteria, 버섯균 등의 linoleate isomerase에 의해 linoleic acid로부터 생합성된다. 그러나 linoleic acid를 알카리 이성화로 합성한 CLA에는 이론적으로 가능한 모든 CLA 이성체가 존재한다. 그 중 c9,t11-CLA와 t10,c12-CLA가 약 45%씩 동일량 존재한다. CLA가 1939 년 linoleic acid의 elaidinization 반응에서 처음으로 소개되었으나 그 이후에는 과학적인 이용가치가 없어 과학자들의 관심의 대상이 되지 않았다. 그러나 1987년에 CLA가 7,12-dimethylbenz[a]anthracene (DMBA)으로 유도한 mouse skin carcinogenesis에서 항암성이 있다는 보고 이후 CLA에 관한 연구는 급속도로 증가하여 현재까지 약 6,100 연구논문이 발표되었다. CLA와 c9,t11-CLA 및 t10,c12-CLA 이성체는 in vitro와 in vivo에서 항암성, 항당뇨성, 항혈압성, 항동맥경화성, 면역증강성, 항산화성, 체지방감소성, testosterone 생산성 등의 생물학적 기능이 갖는다고 밝혀졌다. 이들 생리활성에 c9,t11-CLA와 t10,c12-CLA가 in vitro와 in vivo에서 다른 활성을 보이고 있다. 특히, 합성 CLA에 소량 혼합되어 있는 t,t-CLA가 carcinogen으로 유도한 동물모델이나 암세포에서 다른 이성체 보다 강한 항암성을 보이는 결과는 앞으로 더 많은 연구의 대상이 될 것이다. 본 총설에서는 CLA 관련 연구가 시작된 1939년부터 현재까지의 CLA 연구 트렌드를 살펴보고 밝혀진 주요 기능성을 보고한다. Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid with conjugated double bonds at C9,C11 and C10,C12 positions. Of possible CLA isomers, a naturally occurring CLA isomer is c9,t11-CLA which is produced from linoleic acid by linoleate isomerase from various rumen and lactic bacteria, and mushroom mycelia. Meanwhile, synthetically prepared CLA contained an equal amount of c9,t11-CLA and t10,c12-CLA isomers, and other isomers as minor constituents. CLA was firstly mentioned in 1939 during the elaidinization reaction of linoleic acid. Thereafter, CLA was not an attractant to scientists because it was not scientifically interested any more. However, since the anticarcinogenic action was driven from 7,12-dimethylbenz[a]anthracene (DMBA)-induced mouse skin carcinogenesis in 1987, CLA-related researches were drastically elevated, resulting in approximately 6,100 research papers in literature, so far. CLA exhibited the significant biological activities: anticarcinogenic, antidiabetic, antihypertensive, antiatherosclerotic, body-fat reducing, antioxidative, antiinflammatory, testosterone producing and other activities. Interestingly, two major CLA isomers, c9,t11-CLA and t10,c12-CLA, exhibited different biological activities. Meanwhile, t,t-CLA isomers which is minor constituent of chemically synthesized CLA from linoleic acid exhibited more potent anticarcinogenic activity in carcinogen-induced animal models and cancer cell lines than other CLA isomrs. In the present review, the significant biological activities of CLA were discussed along with historical studies of CLA since 1939.

Ketamine 이 해명(Guinea pig)의 자궁운동에 미치는 영향

김영숙,김종래,안영수,오흥근,최령,홍사석,Robb, Ian S .,김원준 대한마취과학회 1976 Korean Journal of Anesthesiology Vol.9 No.2

Postpartum bleeding of the uterus is an important cause of maternal death. It is thought that the inhibitory effect of the majority of potent inhalation anesthetics on uterine contraction is often responsible (Munson, 1970; Lim et al., 1971; Kim, 1972). There have been numerous reports that ketamine stimulates uterine contraction clinically Chodoff and Stella, 1966; Little et al, 1972; Galbert and Gardner, 1973; Corssen, 1974). Jawa- lekar and associates(1972) reported that ketamine increased resting tension, contractile amplitude nd frequency of the uterine strips of pregnant mice. And Kim(1975) reported research on the offects of ketamine on the isolated uterus of rabbits. According to Kim(1975), ketamine exerted .a stimulatory action on the uterus under the influence of progesterone. This progesterone-depenent uterine stimulatory action of ketamine is not concerned with adrenergic and cholinergic mechanisms but appears to have a direct effect on the uterine muscle. The present study was undertaken to determine whether or not ketamine also exerted a stimlatory action on the uterus of the guinea pig and to follow up whether or not stimulatory action ef ketamine also depends upon progesterone in the guinea pig. Adult female non-pregnant guinea pigs and rabbits weighing approximately 0, 5 kg and 2. 0 kg respectively were employed in this experiment. At the end of 10 to 14 days following bilateral oophorectomy, non-pregnant guinea pigs and rabbits were injected intramusculary with estradiol benzoate (2,000 I.U./kg) or progesterone (Smg/kg) once a day for four consecutive days, A uterine strip, about 1.5~2,0cm in length, was carefully isolated from the experimental animals and suspended in a muscle chamber containing 50 ml of Lockes solution, maintained at constant temperature of 38C. It was aerated with 100% oxygen bubbling through the bathing fluid by means of a sintered glass plate at the bottom of the muscle chamber. One end of the uterine strip was attached to the bottom of the muscle chamber and the other end to a lever. Motility and tonus were recorded on kymograph paper. After being washed several times with fresh Locke's solution, the uterine strip attained a constant motility and tonus. Ketamine then was added in various concentrations to the chamber. The results are as follows: 1. The uterine mctility of guinea pig shows a higher amplitude and lower frequency compared with that of rabbit. 2. Effects of ketamine on isolated uterine strips of guinea pig pre-treated with estrogen showed no change using 1~20 Υ/ml of ketamine, but with 50 Υ/ml or more, the amplitude of uterine contractions was depressed. 3. On isolated uterine strips of guinea pig pre-treated with progesterone, there was no effect with 1100 Υ/ml of ketamine, depressed amplitude of uterine contractions with 150 Υ/ml and markedly depressed amplitude of uterine contraction with 200Υ/ml. 4. On isolated uterine strips of rabbit pre-treated with estrogen, there was no effect with 20Υ/ml of ketamine, but depressed amplitude of uterine contraction u0Υ/ml. 5. On isolated uterine strips of rabbit pre-treated with progesterone, there was an increase in amplitude and frequency of uterine contractions with up to 150Υ/ml of ketamine, but depressed amplitude with 200Υ/ml or more. From the above results it may be concluded that ketamine exerted a stimulatory action on the uterus of rabbit under the influence of progesterone but not on the uterus of guinea pig. The large amount of ketamine depressed the amplitude of uterine contraction in guinea pig and rabbit and this depressing dose is higher in the animals (guinea pig and rabbit) pre-treated with pro- gesterone than with estrogen.

중환자실(I.C.U)환자의 임상통계

김영숙,김종래,박광원,오흥근,정화성,Ian S . Robb,변달섭 대한마취과학회 1976 Korean Journal of Anesthesiology Vol.9 No.2

The concept of intensive care has developed from experience in recovery rooms, in anesthetic work, and in early specialized units in different fields of medicine. Between the First and the Second World Wars special units were created in some German hospitals. Intensive care often means a combination of recovery-room service and intensive therapy. Intensive care, especially respiratory care is an important part of the responsibility of the anesthesiologist. So, we have analyzed statistically 3, 072 I.C.U. patients who,were admitted during a period of 7 years beginning October 18, 1968 when the I.C.U. at Severance Hospital was opened, The results are as follows: I. Cases admitted to the I.C.U. (March,1970-August, 1975 ) 1) For 5 and a half years, the total number of patients was 2, 479. These patients were 2.7% of 91,400 patients who were admitted into Severance Hospital. 2) The mortality rate was 20.0% . Mortality rate was highest in 1970 (23. 7%) and lowest in 1975 (13. 5%). It has gradually decreased every year. 3) Total occupied bed days were 9,840 days (average 4,0 days). The occupied bed days of the majority of the patients (83.6%) were below 5 days comprising 88.3% of the total expired patients. 4) 50% of the patients were in the 21 to 50 year age group. The highest mortality rate occurred in the 51 to 60 age group (24.3%). 5) The number of patients admitted in the department of internal medicine was 702 cases (28.3%) which was the highest among all departments. The mortality rates of the departments of pediatric and neurosurgery were 40. 3% and 34. 7% respectively and those were relatively high values. 6) All I.C.U. patients have been divided into two categories, operative and non-operative. The operative cases were 1, 264 (51. 0%) and non-operative 1, 215 (49. 0%,). The mortality ,The Journal of The Korean Society of Anesthesiologists VoI. 9, No. 2, 1976 rate of the non-operative cases (22. 5%) was higher than operative (17. 6%). 7) Cardio-vascular disease was the commonest cause of death of I.C.U, patients (107 cases) admitted from March, 1974 to August, 1975 (30 cases, 28. 0%). II. Respiratory support cases (Oetober 18, 1968-August 31, 1975) ; 1) During the 7 year period, of all I.C.U. admission cases (3,072) 423 cases (13.8%) recei- ved respiratory support. These patients gradually increased every year. 2) The mortality rate was 44. 0% which was more than twice the over-all mortality rate of. I.C.U. admission patients. 3) The mortality rate was highest in the 21 30 age group (18. 7%). 4) The department of internal medicine admitted the largest number of cases (153 cases, 36. 2%). 5) Of all conditions, cardiovascular disease was most frequent (126 cases, 29. 8%). Here we have dicussed the definition of I.C.U., the responsibility and important role of the anesthesiologist in the I.C.U. by illustrating present situations and presenting problems at the I.C.U. in Severance Hospital.