Rat의 방사선 조사성 구내염에 대한 Recombinant Human Epidermal Growth Factor (rhEGF)의 효과

정권일(Kwonil Jung),김선희(Sunhee Kim),문수영(Soo Young Moon),김연화(Yeon Wha Kim),홍준표(Joon Pio Hong),김현숙(Hyun Sook Kim),이상욱(Sang-wook Lee) 대한방사선종양학회 2006 Radiation Oncology Journal Vol.24 No.1

목 적: 구내염은 두경부종양이 있는 암환자에게 방사선 및 항암제 치료 시술 시 매우 흔하게 발생하는 합병증이다. 본 연구는 Rat의 방사선성 구내염 모델에서 recombinant human epidermal growth factor (rhEGF)의 효과를 평가하 고자 하였다. 대상 및 방법: 25 Gy의 방사선량으로 두부에 단회 조사한 Rat를 무작위로 7마리씩 무처치군, 부형제 처치군, rhEGF 15 또는 30μg/day 구강 내 처치군으로 나누었으며, 방사선을 조사하지 않은 7마리의 Rat를 정상시험군으로 나누었다. rhEGF 시료 및 부형제는 1일 3회 Rat의 구강점막에 매일 도포하였다. Rat의 생존율, 체중변화 및 사료섭 취량을 18일 동안 관찰하였으며, 방사선 조사 후 7일 및 18일째에 Rat의 구강점막을 조직학적으로 평가하였다. 결 과: 실험종료 시점에서 rhEGF 15 또는 30μg/day 구강 내 처치군이 모두 33%의 생존율을 보인 것에 비하여,무처치군 및 부형제 처치군은 모두 0%의 생존율을 보였다. 체중변화에서도 rhEGF 처치군은 방사선 조사 후 2일부터 7일까지 부형제 처치군에 비하여 Rat의 평균체중이 통계적으로 더욱 무거웠다. 사료섭취율은 모든 시험군에서 방사선 조사 후 4일까지 감소하는 양상을 보이다가, rhEGF 15 또는 30μg/day 구강 내 처치군에서 14일째에 뚜렷한 사료섭취율의 증가가 관찰되었다. 방사선 조사 후 7일째의 조직학적 분석 결과, rhEGF 15 또는 30μg/day 구강내 처치군의 Rat에서는 점막 표피층의 각질세포의 종창 및 변성만이 관찰되었던 것에 비하여, 무처치군 및 부형제 처치군에서는 심한 위막성 또는 궤양성 구내염이 관찰되었다. 결 론: rhEGF (15 또는 30μg/day 구강 내 처치군) 처치에서 방사선 조사로 유발시킨 Rat의 구내염 모델에서 유의성 있는 치유 효과를 확인하였으며, 본 시험결과로 rhEGF가 방사선에 의해 유발된 구내염을 치료할 수 있는 임상제제로써의 가능성을 볼 수 있었다. Purpose: Oral mucositis is a common toxicity of radiation or chemotherapy, which is used a treatment for head and neck cancer. We investigated effects of recombinant human epidermal growth factor (rhEGF) on radiation-induced oral mucositis in rat model. Materials and Methods: Spraque-Dawley rats (7 per group) exposed to a single dose of 25 Gy (day 0) on their head, except for one group, were randomly divided into un-treated, vehicle-treated, and two rhEGFtreated groups. Rats were topically applied with rhEGF (15 or 30μg/oral cavity/day) or vehicle to their oral mucosa. Survival rate of rats, weight changes, and food intakes were examined from day 0 to 18 after radiation. Histology study was performed from oral mucosa of rats at day 7 and 18 after radiation. Results: rhEGF-treated groups (15 or 30μg/day) showed all survival rate 33%, whereas un-treated and vehicle-treated groups showed all survival rate 0% at the end of experiment. rhEGF-treated groups statistically had less weight loss compared to vehicle-treated group from day 2 to 7 after radiation. Food intake of rats with rhEGF treatment turned to increase at day 14 after radiation. At 7 day after radiation, un-treated and vehicle-treated groups showed severe pseudomembraneous or ulcerative oral mucositis. On the other hand, rhEGF-treated groups had no more than cellular swelling and degeneration of epidermal cells in oral mucosa of rats. Conclusion: These results suggest that rhEGF has significantly positive effects on radiation-induced oral mucositis in rats. rhEGF display a therapeutic potential on a clinical level.

SD rats의 성별에 따른 형개 열수추출물의 방사선 방호효과

이지은(Ji-Eun Lee),김장오(Jang-Oh Kim),이윤지(Yoon-Ji Lee),전찬희(Chan-hee Jeon),허성회(Sung-Hoe Heo),민병인(Byung-In Min) 한국방사선학회 2021 한국방사선학회 논문지 Vol.15 No.3

본 연구는 형개 열수추출물이 흰 쥐의 성별에 따라 미치는 방사선 방호효과를 평가하는 것이다. 형개는 활성산소를 제거하는 대표적인 물질인 polyphenol과 flavonoid를 함유하고 있다. 이에 따라 방사선 방호효과를 알아보기 위해 형개 열수추출물을 SD rat 수컷과 암컷에 2 주 동안 경구투여 한 후 혈액학적 분석, 소장 융모 길이 변화 분석, SOD 활성 평가를 실시하였다. 수컷 SD rat ST+IR군은 IR군에 비해 혈소판의 회복능력이 조금 더 증가한 것으로 나타났다. 암컷 또한 ST+IR군이 IR군에 비해 혈소판 회복능력이 더 높게 나타났다. 림프구 수치도 혈소판 수치와 마찬가지로 수컷과 암컷 SD rat ST+IR군이 IR군보다 회복능력이 더 높게 나타났다. 방사선 조사 후 수컷 SD rat과 암컷 SD rat의 소장 융모 길이가 IR군보다 ST+IR군이 덜 감소한 것으로 보아 모든 성별의 SD rat 소장 융모가 손상이 적은 것을 확인하였다. 이 결과를 통해 형개 열수추출물이 방사선 방호효과가 있음을 확인 할 수 있었다. This study evaluates the radiation protection effects of Schizonepeta tenuifolia water extract on white rat by gender. Schizonepeta tenuifolia contains polyphenol and flavonoid, which are typical substances that remove free radical. Thus, to determine the effectiveness of radiation protection, the Schizonepeta tenuifolia water extract was administered to Sparaqu-Dawely (SD) rat males and females for two weeks, followed by hematological analysis, analysis of changes in the length of the small intestine villi length, and SOD activity evaluation. The male rat in ST+IR group showed a slightly greater recovery of platelets than the IR group. The ST+IR group also showed a higher platelet recovery capability than the IR group. lymphocyte showed that male and female rat ST+IR groups have higher resilience than IR groups likewise platelet. After irradiation, the villi length of the male rat and female rat decreased less in ST+IR group than in the IR group, indicating that the villi length of all genders was less damaged. This result confirmed that the Schizonepeta tenuifolia water extract had a radiation protection effect.

Human 성장호르몬을 도입한 Transgenic Rats의 작출과 번식표현형에 관한 연구 II. 형질전환된 Rats의 hGH수준이 번식표현형에 미치는 영향

장규태,김성현,성환후,주학진,박미령,윤창현 한국동물번식학회 1998 Reproductive & developmental biology Vol.22 No.2

The effects of continuous GH(hGH) secretion on the female reproduction was studies in adults female transgenic rats expressing the hGH gene with a mouse whey acid protein (mWAP) promotor. Two line of transgenic female rats carrying the mWAP/hGH gene were established and used in the study. One was characterized by relatively high levels of serum hGH (high line), and the other had relatively low levels (low line). 1. High line female rats had recurring, Pseudopregancy-like estrous cycles accompanied by increased serum progesterone level for 2 weeks after ovulation, and they were fertile. 2. In the rats, luteinization occurred spontaneously without cervical stimulation, probably due to high levels of serum hGH, which has prolactin (PRL)-like activity in the rat. 3. Low line female rats had recurring, regular 4-days estrous PRL surge following cervical stimulation were not, detected and PRL secretion was not induced by a dopamine antagonist. 4. The ovarian tissue in this line had a much higher number of corpora lutea and grew much heavier than in normal littermates, suggesting impairment of PRL induced structural luteolized. Su, pp.ession of PRL secretion in the low line rats was, at least in part, due to a marked decrease in the number of lactotrophs in the pituitary. The present study shows that the serum hGH level plays a crucial role in regulating luteal function in female transgenic rats expressing the hGH gene.

WHW®의 랫드에서의 반복경구투여 독성에 관한 연구

오태우,배효상,윤철호,박용기 대한본초학회 2010 大韓本草學會誌 Vol.25 No.3

Objectives : We investigated the repeated-dose toxicity of Wenpitang-Hab-Wulingsan(WHW), a Korean traditional medicine prescribed with twelve herbs, which has been used for the treatment of renal disease. Methods : WHW extract prepared by GLP company. WHW was supplemented by gavage at 0, 100, 500 and 1000 ㎎/㎏/day for 13-week consecutive days. We recorded the clinical signs of toxicity, body weight, organ weights, hematology, gross and histological changes in target organs rats and clinical chemistry analysis for all rats. Results : WHW extract at all doses was shown no mortality or abnormal clinical signs in rats during at the observation period. Furthermore, there was no difference in body weight and food-take consumption, organ weight, gross pathological findings, and urine analysis among the groups of rats treated with different doses of WHW extract. The hematological analysis and clinical blood chemistry data were revealed no toxic effects from WHW-treated rats. Conclusions : The results suggest that WHW extract in rats is a wide margin of safety on a acute toxicity. Objectives : We investigated the repeated-dose toxicity of Wenpitang-Hab-Wulingsan(WHW), a Korean traditional medicine prescribed with twelve herbs, which has been used for the treatment of renal disease. Methods : WHW extract prepared by GLP company. WHW was supplemented by gavage at 0, 100, 500 and 1000 ㎎/㎏/day for 13-week consecutive days. We recorded the clinical signs of toxicity, body weight, organ weights, hematology, gross and histological changes in target organs rats and clinical chemistry analysis for all rats. Results : WHW extract at all doses was shown no mortality or abnormal clinical signs in rats during at the observation period. Furthermore, there was no difference in body weight and food-take consumption, organ weight, gross pathological findings, and urine analysis among the groups of rats treated with different doses of WHW extract. The hematological analysis and clinical blood chemistry data were revealed no toxic effects from WHW-treated rats. Conclusions : The results suggest that WHW extract in rats is a wide margin of safety on a acute toxicity.

90% pancreatectomized rats 에서 w-6 다중불포화지방과 비타민 E 섭취가 인슐린 저항성에 미치는 영향

최수봉(Soo Bong Choi),박선민(Sun Min Park) 대한내과학회 1999 대한내과학회지 Vol.56 No.4

N/A High intake of -6 polyunsaturated fat (PUFA) can increase oxidative stress, which may in turn increase insulin resistance and be the cause of metabolic syndrome X (diabetes mellitus). One of the ways to reduce oxidative stress is through the consumption of antioxidants, such as vitamin E. However, it is controversial whether the consumption of vitamin E alleviates insulin resistance. The purpose of this study was to determine whether the excessive intake of -6 polyunsaturated fat and vitamin E influences the whole body glucose disposal rate (GDR), glycogen deposits in the liver and muscles, and the triglyceride content of muscles in 90% pancreatectomized rats. Methods : Mildly diabetic rats were produced by removing 90% of the pancreas from Sprague Dawley rats aged 8 weeks. One week after surgery, the blood glucose levels of rats were more than 9.4 mmol/L, according to which the rats are considered to be diabetic. Two factors were examined in the 90% pancreatectomized rats: polyunsaturated fat levels with 40% and 10% of total energy intakes, and vitamin E contents with 300 IU and 30 IU per kg of diet mixture. Four different diets were given for 8 weeks. After 7 weeks of diet consumption indwelling catheters were inserted in the carotid artery and jugular vein of all rats so that the GDR could be measured while the rats were awake and in a relaxed state. Results : Daily calorie and PUFA fat intake levels were higher in the high PUFA groups of diabetic rats. Daily vitamin E intake of the high vitamin E groups was ten times higher than that of the low vitamin E groups (p<0.0001). The GDR was lower in high PUFA groups, and the high intake of vitamin E tended to decrease the GDR in diabetic rats. The GDR of DHPHE was significantly lower than that of DLPLE (p<0.05). Sham operated rats (the control group) had a higher GDR by 44% than diabetic rats. Vitamin E deposits in the liver in DHPHE were higher in DLPLE (p<0.05). Glycogen deposits in the liver of diabetic rats were significantly higher in DLPLE than DHPLE. Muscle glycogen content showed a similar tendency to liver glycogen content in different diet groups of diabetic rats. Triglyceride deposits in muscles did not differ according to dietary fat and vitamin E contents in diabetic rats. Conclusions : High intake of -6 PUFA increased insulin resistance, and high vitamin E intake did not alleviate insulin resistance. Thus, increased oxidative stress may be an independent factor for increased insulin resistance in high intake of -6 PUFA. It is necessary to consume a minimum amount of -6 PUFA as well as the Recommended Daily Allowance of vitamin E.

고혈압쥐 노화 적혈구에서의 Na, K-ATPase에 관한 연구

박창길(Chang-Kil Park),허강민(Gang-Min Hur),석정호(Jung-Ho Seok),이재흔(Jae-Heun Lee) 대한약리학회 1991 대한약리학잡지 Vol.27 No.1

고혈압백서(1-kidney, 1-clip-hypertensive rat)의 적혈구에서 노화 과정에 따른 Na, K-ATPase의 변동을 관찰하고자 노화적혈구를 분리한 다음 세포막에서의 Na-pump 활성도 및 ouabain의 결합실험과 Rb의 세포내 유입실험을 시행하여 다음과 같은 결과를 얻었다. 1. 본 실험에 사용한 고혈압 백서의 혈압은 수축기 및 이완기 혈압이 165.5/119.0 mmHg로 유의하게 증가 하였다. 노화 적혈구의 평균용적(MCV)과 세포막 단백질 함량은 감소되고 혈색소치는 증가되었다. 2. 110 mM NaCl 및 10 mM KCI 존재하에서의 적혈구 세포막 Na, K-ATPase활성도는 대조군에 비해 고혈압군에서 억제 되었으며 양군 모두에서 노화에 의해 그 활성도가 감소되었다. 3. 4 mM RbCl 존재 하에서 Ouabain에 의해 억제되는 Rb의 유입은 정상 및 고혈압군의 노화적혈구에서 약간 감소되었으며 고혈압군의 young erythrocyte에서는 오히려 약간 증가 되었다. 4. 16 mM RbCl 존재 하에서 Ouabain에 의해 억제되는 Rb의 유입은 양군의 노화 적혈구에 서는 각군의 young erythrocyte에 비해 약 30-50% 감소되었으며, 고혈압군에서는 특히 young erythrocyte에서 정상군의 young erythrocyte에 비해 유의하게 감소되었다. 5. 0.13 × 10^-6M과 1 × 10^-6M에서의 ouabain binding은 정상군의 노화적혈구에서는 young erythrocyte에 비해 약간 감소되었으나 고혈압군의 노화적혈구에서는 유의하게 감소되었다. 6. 6 × 10^-6M과 64 × 10^-6M 에서의 ouabain binding은 양군의 노화 적혈구에서는 약간 감소되었지만 유의성은 없었으며 고혈압군의 young erythrocyte 및 노화적혈구에서는 정상군의 young erythrocyte및 노화 적혈구에 비해 유의하게 감소되었다. 이상의 결과로부터 (1) 고혈압쥐의 young erythrocyte에서는 low affinity의 Na-pump수의 감소 및 molecular activity의 증가, (2) 정상쥐의 노화 적혈구에서는 molecular activity의 저하, (3) 고혈압쥐의 노화적혈구에서는 molecular activity의 저하 및 high affinity와 low affinity의 Na-pump수의 저하등에 의하여 Na-pump의 기능이 변동될 수 있을 것으로 추측된다. To study the age dependent change of Na, K-ATPase in the erythrocyte of hypertensive rat, 1-kidey 1-clip hypertensive rat was made by the removal of right kidney and partial ligation of left renal artery. After 4 weeks, aged erythrocyte fraction was separated by density gradient centrifugation, and Na, K-ATPase activity and ³H-ouabain binding with ghost cell membrane and ouabain sensitive Rb-uptake with whole cell were measured. 1) In the hypertensive rats, blood pressure was significantly increased to 165.5/119.0 mmHg (systolic/diastolic). Mean corpuscular volume and membrane protein(mg) per 10⁹RBC were decreased and hemoglobin content was increased in the aged erythrocyte. 2) Na, K-ATPase activity in the solution containing 110 mM NaCl and 10 mM KCI, was decreased in hypertensive rat, and decreased in aged erythrocyte of both group. 3) Ouabain sensitive Rb-uptake by low RbCl concentration(4 mM) was slightly decreased in aged erythrocyte compared to that in young erythrocyte of each group, but slightly increased in young erythrocyte in hypertensive rat compared to that in normotensive rat. 4) Ouabain sensitive Rb-uptake by high RbCl concentration(16 mM) was decreased about 30% to 50 % in aged erythrocyte in both group. And in hypertensive rat, especially in young erythrocyte it was significantly decreased compared to that in normotensive rats. 5) ³H-ouabain binding at 0.13 or 1 × 10^-6M ouabain concentration was slightly decreased in aged erythrocyte of normotensive rat, and significantly decreased in aged erythrocyte of hypertensive rats. 6) ³H-ouabain binding at 6 or 64 × 10^-6M ouabain concentration is slightly decreased in aged erythrocyte of both group, but significantly decreased in young and aged erythrocyte of hypertensive rats compared to that of normotensive rats. The present results suggest that (1) in the young erythrocyte of hypertensive rat, the alterations of Na-pump activity that slightly increased in weak stimulation and inhibited in strong stimulation, may be related to increased molecular activity and the decrease in the number of low affinity site without change in high affinity site, (2) in the aged erythrocyte of normotensive rat, inhibited Na-pump may be related to the change in molecular activity of pump. (3) And in the aged erythrocyte of hypertensive rat, it may be related to the decrease in the number of high and low affinity site as well as the change in molecular activity.

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.

Comprehensive Evaluation of Models for Studying Obesity

Jun-Won Yun,Byoung-Seok Lee,Jin-Suk Hyun,Chae-Wook Kim,Dong-Wook Shin,Tae-Ryong Lee,Bae-Hwan Kim 한국실험동물학회 2005 Laboratory Animal Research Vol.21 No.3

Obesity is a serious risk factor for so-called lifestyle-related diseases such as diabetes, cardiovascular disease and hypertension. Although obesity is caused by various environmental and genetic factors, the consumption of high-fat diet (HFD) is thought to be one of the main factors. This study was designed to evaluate two rat models, Zucker model and HFD-fed Sprague-Dawley (SD) model, for studying obesity. The order of epididymal fat pad weights were as follows: Zucker obese rats (0.025±0.003) > HFD-fed SD rats (0.016±0.004) > Normal diet (ND)-fed SD rats (0.012±0.001) > Zucker lean rats (0.011±0.000). Zucker obese group and HFD-fed SD group showed increasing tendency in blood glucose concentrations. On the other hand, serum triglyceride levels, free fatty acid (FFA) levels and cholesterol levels were significantly higher in only Zucker obese rats compared with other groups. The histological examination of the liver tissue from Zucker obese rats revealed an altered liver structure with a huge macrovacuolar steatosis disseminated from the portal space to the centrolobular vein. In HFD-fed SD rats, enlarged hepatocytes with microvacuoles were irregularly disseminated. There was also significant increment in serum aspartate transaminase (AST) levels in HFD-fed SD rats, associated with accumulation of lipid in liver. While mineralization of renal tubule and enlargement of pancreatic islet were prominent features of Zucker obese rats, these changes were not seen in HFD-fed SD rats and in Zucker lean rats. Subcutaneous triglyceride levels of Zucker obese rats were statistically higher than those of other groups. These results supported that Zucker obese rat is a useful model of obesity research, as reported previously. On the other hand, triglyceride levels in subcutaneous adipose tissue of HFD-fed SD rats were not significantly different from those of other groups. These results suggested that HFD-fed SD rat model is not suited for investigation of lipolytic effect in subcutaneous adipose tissue. However, HFD-fed SD rats were induced slight obesity compared with Zucker obese rats. Although there were no significant effects from several analytic factors, HFD-fed SD rat model may be a useful tool for studying the mechanisms by which dietary fat influences the regulation of energy balance. Recently, numerous studies on obesity using rodents feeding of a various type of HFD have been preceded. Hence, further studies, considering various feeding duration, different diet (ND and HFD) types and fat content, may be necessary to establish useful animal model for HFD-induced obesity and diabetes.

칼로리량 급여 제한이 OLETF 흰쥐의 콜레스테롤량에 미치는 영향

박재윤,김태형,이병래,차종희 조선대학교 부설 의학연구소 2002 The Medical Journal of Chosun University Vol.27 No.1

본 실험은 식이 제한 급여에 따른 비만형 당뇨병 환자의 체중 및 혈중 지질량의 변화에 대한 연구의 일환으로 사람의 비만형 당뇨병과 유사한 병리소견을 나타내는 OLETF 흰쥐를 이용하여 칼로리 급여량 비 제한군, 칼로리 급여량 20% 제한군과 40% 제한군으로 나누어 12주간 사육한 후 체중과 혈당, 혈액 cholesterol 및 중성지질량을 측정하여 칼로리 급여량제한에 따른 혈액 지질량의 변화를 관찰하였다. 체중은 칼로리 급여량 20% 제한군에서는 체중증가 억제 비율이 LETO와 OLETF흰쥐가 비슷하였고, 칼로리 급여량 40% 제한군에서는 OLETF 흰쥐에서 체중 증가 억제가 더 크게 나타났다. 칼로리 급여량 제한에 의한 혈당량의 저하는 비만형 당뇨쥐인 OLETF 증 흰쥐가 LETO 흰쥐에 비하여 더 크게 나타났다. 칼로리 급여량 제한으로 OLETF 종 흰쥐에서 혈액 중성지질량, 총 콜레스테롤량 및 LDL- 콜레스테롤량이 감소되었고, HDL-콜레스테롤량은 증가되므로, 비만형 당뇨병환자에서 칼로리 제한 식사는 OLETF 종 흰쥐에서와 유사한 혈액 콜레스테롤량의 변화를 유발시켜서 혈관 합병증을 억제할 수 있을 것으로 예상된다. 이상의 실험결과로서 칼로리 급여량 제한에 의한 체중, 혈당, 혈액 총 콜레스테롤량, LDL-콜레스테롤 및 HDL-콜레스테롤량의 변화가 비당뇨 흰쥐보다 당뇨흰쥐에서 더 큰 변화가 있음이 확인되어 칼로리 제한 급여는 당뇨병에서 체중, 혈당, 콜레스테롤량의 조절에 유의한 것으로 생각되고, 칼로리 급여 20% 제한군과 40% 제한군 사이에서 체중, 혈당, 콜레스테롤량의 변화는 칼로리 제한량의 변화에 비하여 적기 때문에 적절한 체중, 혈당, 혈중 지질량의 유지를 위해서는 칼로리 급여량을 적절히 유지하는 것이 중요할 것으로 생각된다. 또한 칼로리 급여 제한에 의한 체중, 혈당, 혈중 중성지질과 콜레스테롤량의 감소는 혈관 합병증 발생이 예방 할 수 있을 것으로 추측되는데, 이에 대해서는 계속적인 연구가 있어야 할 것으로 생각된다. Objective: To determine the effects of caloric restriction on blood cholesterol levels of obese type 2 diabetes, we measured blood glucose and cholesterol level in Otsuka Long Evans Tokushima Fatty(OLETF) rats after caloric restriction. Methods: OLETF rats(obese diabetic rats) and Long Evans Tokushima Otsuka(LETO) rats(nondiabetic control rats) were grouped into 3 groups, control group, 20% caloric restriction(20% CR) group and 40% caloric restriction(40% CR) group, respectively. The blood glucose and cholesterol level in OLETF and LETO rats were determined after 12 weeks caloric restriction. The levels of triglyceride(TG) and cholesterol(total cholesterol, LDL-cholesterol, HDL-cholesterol) in blood of rats were determined by automatic biochemical analyser(Hitachi 747 Japan). Results: Blood glucose level of LETO rats were decreased by 12% on 40% CR compared to control group but the differences of blood glucose levels between control group and 20% CR group was not observed. OLETF rats blood glucose levels were decreased by 20% in 40% CR group and by 15% in 20% CR group. The levels of blood triglyceride in 20% CR and 40% CR groups were decreased by 20%, 15% in LETO rats and by 37%, 32% in OLETF rats respectively. Total cholesterol level was not changed by dietary restriction in LETO rats, but in OLETF rats, total cholesterol levels was decreased significantly by caloric restriction. The HDL-cholesterol levels were increased while LDL-cholesterol levels were decreased by dietary restriction in both LETO and OLETF rats. Conclusion: These results suggested that OLETF rats may be more susceptive to changes of body weight and blood glucose, triglyceride and cholesterol levels than LETO rats by caloric restriction. Reduced LDL-cholesterol and triglyceride levels in diabetes by caloric restriction may have some roles on prevention of diabetic vascular complication.

Na, K-ATPase Activity in the Aged Erythrocytes of Hypertensive Rats

박창길,허강민,석정호,이재흔,Park, Chang-Kil,Hur, Gang-Min,Seok, Jung-Ho,Lee, Jae-Heun The Korean Society of Pharmacology 1991 대한약리학잡지 Vol.27 No.1

고혈압백서(1-kidney, 1-clip-hypertensive rat)의 적혈구에서 노화 과정에 따른 Na, K-ATPase의 변동을 관찰하고저 노화적혈구를 분리한다음 세포막에서의 Na-pump 활성도 및 ouabain의 결합실험과 Rb의 세포내 유입실험을 시행하여 다음과 같은 결과를 얻었다. 1. 본 실험에 사용한 고혈압 백서의 혈압은 수축기 및 이완기 혈압이 165.5/119.0 mmHg로 유의하게 증가 하였다. 노화 적혈구의 평균용적(MCV)과 세포막 단백질 함량은 감소되고 혈색소치는 증가되었다. 2. 110 mM NaCl 및 10 mM KCI 존재하에서의 적혈구 세포막 Na, K-ATPase활성도는 대조군에 비해 고혈압군에서 억제 되었으며 양군 모두에서 노화에 의해 그활성도가 감소되었다. 3. 4 mM RbCl존재하에서 Ouabain에 의해 억제되는 Rb의 유입은 정상 및 고혈압군의 노화적혈구에서 약간 감소되었으며 고혈압군의 young erythrocyte에서는 오히려 약간 증가 되었다. 4. 16 mM RbCl 존재하에서 Ouabain에 의해 억제되는 Rb의 유입은 양군의 노화 적혈구에 서는 각군의 young erythrocyte에 비해 약 30-50% 감소되었으며, 고혈압군에서는 특히 young erythrocyte에서 정상군의 young erythrocyte에 비해 유의하게 감소되었다. 5. $0.13{\times}10^{-6}M$과 $1{\times}10^{-6}M$에서의 ouabain binding은 정상군의 노화적혈구에서는 young erythrocyte에 비해 약간 감소되었으나 고혈압군의 노화적혈구에서는 유의하게 감소되었다. 6. $6{\times}10^-6}M$과 $64{\times}10^-6M$ 에서의 ouabain binding은 양군의 노화 적혈구에서는 약간 감소되었지만 유의성은 없었으며 고혈압군의 young erythrocyte 및 노화적혈구에서는 정상군의 young erythrocyte및 노화 적혈구에 비해 유의하게 감소되었다. 이상의 결과로부터 (1) 고혈압쥐의 young erythrocyte에서는 low affinity의 Na-pump수의 감소및 molecular activity의 증가, (2) 정상쥐의 노화 적혈구에서는 molecular activity의 저하, (3) 고혈압쥐의 노화적혈구에서는 molecular activity의 저하 및 high affinity와 low affinity의 Na-pump수의 저하등에 의하여 Na-pump의 기능이 변동될 수 있을 것으로 추측된다. To study the age dependent change of Na, K-ATPase in the erythrocyte of hypertensive rat, 1-kidey 1-clip hypertensive rat was made by the removal of right kidney and partial ligation of left renal artery. After 4 weeks, aged erythrocyte fraction was separated by density gradient centrifugation, and Na, K-ATPase activity and $^3H-ouabain$ binding with ghost cell membrane and ouabain sensitive Rb-uptake with whole cell were measured. 1) In the hypertensive rats, blood pressure was significantly increased to 165.5/119.0 mmHg (systolic/diastolic). Mean corpuscular volume and membrane protein(mg) per $10^9RBC$ were decreased and hemoglobin content was increased in the aged erythrocyte. 2) Na, K-ATPase activity in the solution containing 110 mM NaCl and 10 mM KCI, was decreased in hypertensive rat, and decreased in aged erythrocyte of both group. 3) Ouabain sensitive Rb-uptake by low RbCl concentration(4 mM) was slightly decreased in aged erythrocyte compared to that in young erythrocyte of each group, but slightly increased in young erythrocyte in hypertensive rat compared to that in normotensive rat. 4) Ouabain sensitive Rb-uptake by high RbCl concentration(16 mM) was decreased about 30% to 50 % in aged erythrocyte in both group. And in hypertensive rat, especially in young erythrocyte it was significantly decreased compared to that in normotensive rats. 5) $^3H-ouabain$ binding at 0.13 or $1{\times}10^-6M$ ouabain concentration was slightly decreased in aged erythrocyte of normotensive rat, and significantly decreased in aged erythrocyte of hypertensive rats. 6) $^3H-ouabain$ binding at 6 or $64{\times}10^-6M$ ouabain concentration is slightly decreased in aged erythrocyte of both group, but significantly decreased in young and aged erythrocyte of hypertensive rats compared to that of normotensive rats. The present results suggest that (1) in the young erythrocyte of hypertensive rat, the alterations of Na-pump activity that slightly increased in weak stimulation and inhibited in strong stimulation, may be related to increased molecular activity and the decrease in the number of low affinity site without change in high affinity site, (2) in the aged erythrocyte of normotensive rat, inhibited Na-pump may be related to the change in molecular activity of pump. (3) And in the aged erythrocyte of hypertensive rat, it may be related to the decrease in the number of high and low affinity site as well as the change in molecular activity