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Developmental competence and Effects of Coculture after Crypreservation of Blastomere-Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis (착상 전 유전진단 기술 개발의 동물실험 모델로서 할구 생검된 생쥐 배아에서 동결보존 융해 후 배아 발생 양상과 공배양 효과에 관한 연구)

  • Kim, Seok-Hyun;Kim, Hee-Sun;Ryu, Buom-Yong;Choi, Sung-Mi;Pang, Myung-Geol;Oh, Sun-Kyung;Jee, Byung-Chul;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Chae, Hee-Dong;Kim, Chung-Hoon
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.47-57
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    • 2000
  • Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

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Effect of Pinus densiflora Extract on Blood Glucose Level, OGTT and Biochemical Parameters in Streptozotocin Induced Diabetic Rats (솔잎증류액의 투여가 Streptozotocin으로 유도한 당뇨쥐에서 혈당, 구강내당능검사, 혈액 성분에 미치는 영향)

  • Kim, Shin-Hee;Hwang, Seock-Yeon;Park, Oh-Sung;Kim, Moo-Kang;Chung, Young-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.973-979
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    • 2005
  • To investigate the effect of Pinus densiflora on biochemical parameters in type I diabetic rats, we evaluated the changes of body weight, fasting blood glucose level, oral glucose tolerance test (OGTT) and biochemical parameters after the intraperitoneal injection of distilled solution of Pinus densiflora in streptozotocin (STZ)­induced rats. Thirty-seven male Sprague Dawley rats $(180\pm10g)$ were divided into four groups; diabetic mellitus (DM) group received STZ (50 mg/kg BW, i.v.); low level of pine extract (LP) group received Pinus densiflora (5 mg/kg BW, i.p.), high level of pine extract (HP) group received Pinus densiflora (10 mg/kg BW, i.p.) after the single injection of STZ (50 mg/kg BW, i.v.), respectively. Normal control (NC) group received saline. The change of fasting blood glucose level and OGTT were measured using glucocard II, and the change of biochemical parameter were measured by Automatic Chemistry Analyzer (Hitach-747, Japan). Mean body weight change of DM group was retarded greatly by STZ-exposure. While, body weights of LP and HP groups were progressively increased with some fluctuation, although the increase rates were slower than that of NC group. Fasting blood glucose levels of LP and HP groups were reduced by Pinus densiflora injection, although the fasting blood glucose levels were higher than that of NC group. The results of OGTT was significantly improved in both of LP and HP group compared to DM group. Increases of blood glucose, alanine aminotransferase (ALT), alkaline phosphatase (ALP) and blood urea nitrogen (BUN) levels by STZ-exposure were attenuated by the Pinus densiflora treatment (p<0.05). From the results, it was suggested that Pinus densiflora has a tendency to decrease STZ-induced toxicity in terms of monitoring fasting blood glucose, OGTT and some biochemical parameters of rat.

Isocyanate-Induced Occupational Asthma: Immunologic and Challenge Studies (Isocyanate에 의한 직업성 천식환자에서 임상양상과 Isocyanate 특이 IgE 항체에 관한 연구)

  • Suh, Jung-Eun;Park, Hae-Sim;Kim, Seong-Jin;Rhu, Nam-Soo;Cho, Dong-Ill;Kim, Jae-Won;Kyung, Nan-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.490-501
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    • 1992
  • Background: Isocyanate is the most significant cause of occupational asthma in this country. The mechanism of isocyanate induced bronchoconstriction is unclear. Subjects and Method: To observe its immunologic and clinical findings, we performed methacholine bronchial challenge test (MBCT), toluene diisocyanate (TDI)-bronchoprovocation test (BPT) and RAST to TDI-, diphenylmethane diisocyanate (MDI)-, and hexamethylene diisocyanate (HDI)-human serum albumin (HSA) conjugate in 22 isocyanate-sensitive asthmatic workers. Results: BPT revealed early (11), dual (5), and late only (6) asthmatic responses. Their latent period ranged from 3 to 120 months (mean:45.9 months). Three cases (13.6%)showed a negative response on initial MBCT, but following MBeT performed 24 hours after TDI-BPT revealed the development of airway hyperresponsivenss. Twelve (54.5%) workers had increased specific IgE to TDI-HSA, seven (31.8%) had to MDI-HSA, and nine (40.9%) had to HDI-HSA conjugate. The prevalence of specific IgE was not associated with latent period, type of asthmatic responses, smoking, and atopic status. After 3 months' avoidance from workplace, airway hyperresponsiveness was improved in 10 (38.3%), among 12 followed cases. Conclusion: It is suggested that isocyanate can induce IgE-mediated bronchoconstriction in 59.1% of isocyanate-sensitive asthmatic workers. Isocyanate-induced asthma can occur even though MBeT showed a negative result, and measurement of the changes of airway hyperresponsivenss after isocyanate-BPT could be helpful to diagnose isocyanate-sensitive asthma.

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IN VITRO COMPARISON OF VARIOUS DIAGNOSTIC METHODS OF OCCLUSAL CAR10US LESIONS (교합면 우식병소의 다양한 진단법에 관한 비교연구)

  • Kim, Jae-Gon;Kim, Young-Jin;Kim, Young-Sin;Baik, Byeong-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.4
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    • pp.613-619
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    • 2001
  • The aims of this study were to compare the accuracy, sensitivity and specificity of cnventional visual examination, radiography and a new laser fluorescence method, KaVo Diagnodent, for the detection of occlusal caries lesions. One hundred sound human premolars and molars which had no restorations or interproximal cavities were tested by three methods. Tooth lesions depth was assessed at histologic examination using Caries detector dye The following results were obtained. 1. Diagnodent show 7.8 in sound tooth, 25.4 in initial caries, 30.5 in enamel caries, and 53.8 in dentin caries with average score 2. Spearman and Pearson relation coefficient was high between tooth-specimen test with dye and Diagnodent(0.736, 0.619), visual examination(0.664, 0.666), and was low between tooth-specimen test with dye and radiographic examination(P<0.01, total) 3. Accuracy of occlusal caries was highest on Diagnodent(65%) and lowest on radiographic examination(35%) 4. In initial caries, the sensitivity and specificity of Diagnodent method was the highest. In enamel caries, the sensitivity of visual examination was the highest and specificity of Diagnodent method was the highest. In dentinal caries, the sensitivity and specificity of Diagnodent method was the highest and sensitivity of visual examination was the lowest.

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Phospholipase C-γ Activation by Direct Interaction with β-Tubulin Isotypes (베타 튜불린에 의한 포스포리파제 C-감마1의 활성화)

  • Lee, In-Bum;Kim, Sung-Kuk;Choi, Jang-Hyun;Suh, Pann-Ghill;Chang, Jong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.612-617
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    • 2006
  • Phosphoinositide-specific phospholipase $C-{\gamma}\;1\; (PLC-{\gamma}\;1)$ has pivotal roles in cellular signaling by producing second messengers, inositol 1,4,5-trisphosphate $(IP_3)$ and diacylglycerol (DG). Tubulin is a main component of microtubules and mitotic spindle fibers, which are composed of ${\alpha}-$ and ${\beta}-tubulin$ heterodimers in all eukaryotic cells. In humans, six ${\beta}-tubulin$ isotypes have been identified which display a distinct pattern of tissue expression. Previously we found that $PLC-{\gamma}\;1$ and one of four ${\beta}-tubulin$ isotypes including ${\beta}1$, ${\beta}2$, ${\beta}3$ and ${\beta}6$, colocalized in COS-7 cells and cotranslocated to the plasma membrane to activate $PLC-{\gamma}\;1$ upon agonist stimulation. In the present study, we demonstrate that the remaining two, tubulin ${\beta}4$ and ${\beta}5$, also showed a potential to activate $PLC-{\gamma}\;1$. The phosphatidylinositol 4,5-bisphosphate $(PIP_2)$ hydrolyzing activity of $PLC-{\gamma}\;1$ was substantially increased in the presence of purified ${\beta}4$ and ${\beta}5$ tubulin in vitro, whereas the activity was not promoted by bovine serum albumin, suggesting that tubulin ${\beta}4$ and ${\beta}5$ also activate $PLC-{\gamma}\;1$. Taken together, our results suggest that all the ${\beta}-tubulin$ isotype activates $PLC-{\gamma}\;1$ activity to regulate cellular signaling.

Effect of Liquid Cultures of Cordyceps militaris on Lipid Metabolism and Enzyme activities in hyperlipidemic Female Rats (동충하초 균사체 액체배양액이 고지혈증 흰쥐의 지질대사 및 효소활성에 미치는 영향)

  • 고진복
    • Journal of Life Science
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    • v.13 no.3
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    • pp.265-272
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    • 2003
  • The effect of liquid cultures of Cordyceps militaris (LCM) on weight gain, food intakes, food efficiency ratios, serum and hepatic lipids, fecal lipids excretion, serum protein and enzyme activities, were investigated in adult female rats (30 weeks old). Sprague-Dawley rats were assigned to one normal and four hyperlipidemic diet groups, Hyperlipidemic diet groups (20% fat, 1% cholesterol) were divided into high fat diet (LCM free water), 10%, 20% or 30% LCM diet groups (10%, 20% or 30% LCM in water) according to the levels of LCM supplementation. After 35 days of experimental diet consumption, the body weight gains, hepatic weights, and food efficiency ratios of the rats fed hyperlipidemic diets were significantly increased compared with those of the rats fed normal diet. The concentrations of serum and hepatic triglycerides, hepatic total lipid, and atherogenic index of the rats fed 20% or 30% LCM diets were significantly lower than those of the rats fed the high fat diet. The concentration of serum HDL-cholesterol of the rats fed all LCM diets was significantly higher than those of the rats fed the high fat diet. The fecal excretion of triglyceride in the rats fed 20% or 30% LCM diets was significantly higher than those of the rats fed high fat diet. The concentrations of serum and hepatic total cholesterol, serum LDL-cholesterol, and HDL-cholesterol/total cholesterol ratio, fecal excretion of cholesterol, and the activities of serum glutamic pyruvic transaminase and alkaline phosphatase of the rats fed all LCM diets were similar to those of the rats fed high fat diet. No differences were noted in the weights of kidney and femur, the serum concentration of glucose, total protein and albumin, and the activities of glutamic oxaloacetic transaminase and ${\gamma}$ -glutamyltranspeptidase, among the rats on all the experimental diets. These results showed that the 20% or 30% LCM diets feeding decreased the serum and hepatic triglycerides, and the atherogenic index, and increased the serum HDL-cholesterol of the adult female rats.

Effect of Black Garlic and Herb Formulas on Lipid Profiles and Antioxidant Status in Rats by Interval Running Training (간헐적 운동부하에서 흑마늘과 생약재의 복합처방이 흰쥐의 체내 지질 성분 및 항산화에 미치는 영향)

  • Lee, Soo Jung;Kim, In Sung;Lee, Hye Jin;Oh, Soo Jeong;Shin, Jung Hye;Kim, Jeong Gyun;Sung, Nak Ju
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1436-1444
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    • 2013
  • To improve the functionality of black garlic drinks, black garlic extract (5%) and five herb extracts (1%) were mixed in 70:30 (v/v) ratios as BHF1, and BHF2 was prepared using a 3X concentration of BHF1. After the black garlic and herb formulas (BHFs) were administered over the course of five weeks in rats by interval running training, the lipid profiles and the antioxidant enzyme activities were tested. The total phenolic content of the BHFs were significantly higher in BHF2 than they were in BHF1, and their antioxidant activities were dependent upon the total phenolic content. No significant difference was found in the total serum protein levels among the rats in the Ex-con group by interval running training and the rats in the BHFs-fed groups. However, the albumin level was significantly higher in the Ex-BHF2 to Ex-con group. AST and ALT activities significantly decreased in the BHFs-fed groups compared to the Ex-con group. In terms of changes in the serum lipid profiles, no significant difference was found between the specimens that underwent interval running training and those that did not undergo interval running training. Triglyceride levels, total cholesterol, LDL-C, and HTR levels in the serum were significantly decreased in the Ex-BHF2 to Ex-con group. No significant difference was found in the total lipid levels in the livers of the BHFs-fed groups and the Ex-con group. The triglyceride levels and total cholesterol levels in the Ex-BHF2 group were significantly lower compared to another group. Hepatic catalase activity was significantly increased in the Ex-BHF2 group, but SOD and GSH-px activities were significantly increased as the concentration of the BHF. The antioxidant enzyme activities by supplementation of BHFs increased; thus, three intakes of BHF each day could improve antioxidant status against different types of oxidative stress.

Factors Predicting the Development of Radiation Pneumonitis in the Patients Receiving Radiation Therapy for Lung Cancer (방사선 치료를 시행 받은 폐암 환자에서 방사선 폐렴의 발생에 관한 예측 인자)

  • An, Jin Yong;Lee, Yun Sun;Kwon, Sun Jung;Park, Hee Sun;Jung, Sung Soo;Kim, Jin whan;Kim, Ju Ock;Jo, Moon Jun;Kim, Sun Young
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.1
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    • pp.40-50
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    • 2004
  • Background : Radiation pneumonitis(RP) is the major serious complication of thoracic irradiation treatment. In this study, we attempted to retrospectively evaluate the long-term prognosis of patients who experienced acute RP and to identify factor that might allow prediction of RP. Methods : Of the 114 lung cancer patients who underwent thoracic radiotherapy between December 2000 and December 2002, We performed analysis using a database of 90 patients who were capable of being evaluated. Results : Of the 44 patients(48.9%) who experienced clinical RP in this study, the RP was mild in 33(36.6%) and severe in 11(12.3%). All of severe RP were treated with corticosteroids. The median starting corticosteroids dose was 34 mg(30~40) and median treatment duration was 68 days(8~97). The median survival time of the 11 patients who experienced severe RP was significantly poorer than the mild RP group. (p=0.046) The higher total radiation dose(${\geq}60Gy$) was significantly associated with developing in RP.(p=0.001) The incidence of RP did not correlate with any of the ECOG performance, pulmonary function test, age, cell type, history of smoking, radiotherapy combined with chemotherapy, once-daily radiotherapy dose fraction. Also, serum albumin level, uric acid level at onset of RP did not influence the risk of severe RP in our study. Conclusion : Only the higher total radiation dose(${\geq}60Gy$) was a significant risk factor predictive of RP. Also severe RP was an adverse prognostic factor.

Guidelines of Bioequivalence Studies of Medical Products in Europe (유럽의 의약품 생물학적 동등성 시험 가이드라인)

  • Yoo, Tae-Moo;Yi, Sun-Woo;Park, In-Sook;Suh, Soo-Kung;Ahn, Mee-Ryung;Choi, Hong-Suk;Jin, Sook;Sohn, Soo-Jung;Yang, Ji-Sun
    • Journal of Pharmaceutical Investigation
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    • v.30 no.4
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    • pp.299-307
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    • 2000
  • 생물학적 동등성 시험은 동일 성분을 동량 함유한 제제가 유사조건에서 투여되었을 경우 그 유효성분이 전신순환혈이나 작용부위에서 유용하게 되는 속도 및 양에 큰 차이가 없음을 입증하는 시험이다. 즉 이미 제조허가 되어 시판중인 의약품과 동일한 품목의 제조허가를 받기 위한 경우이거나 생체이용률이 동일함을 증명하기 위해 실시하는 시험이다. 우리 나라에서는 1989년 1월 1일 이후 허가된 전문의약품으로서 신약과 동일한 의약품, 패취제제, 제제개선을 통해 흡수율을 높이는 경우에 생물학적 동등성 시험을 하도록 의무화하고 있다. 미국 FDA는 1975년 6월 생체이용률, 생물학적 동등성에 관한 규제안을 발표하였고, 우리 나라에서도 1988년 10월에 생물학적 동등성 시험 기준을 제정하였다. 유럽에서는 1991년 12월 CPMP(Committee for Proprietary Medicinal Products)에 의해 최초로 "생체이용률 및 생물학적 동등성에 대한 가이드라인"이 채택되었다. 그후 전문가 회의를 거쳐 규정 전반에 관한 것과 일반적 항목에 대한 검토가 이루어져 최종 개정된 가이드라인이 만들어졌다. 현재 in vivo 생체 이용률을 비교하는 생물학적 동등성 시험의 중요성이 증대되고 있고, 국제적으로도 각 국가별 생물학적 동등성 시험의 평가방법이나 기준설정에 대한 연구가 다양하게 이루어지고 있다. 또한 ICH에서도 생물학적 동등성 시험의 국제적 조화를 위한 시도가 이루어지고 있다. 우리 나라의 현행 생물학적 동등성 기준은 1998년 개정된 것을 사용하고 있으나 국제적 발전 추세에 맞추어 기준의 제고를 모색하고 있다. 그 일환으로 유럽의 "의약품의 생체이용률 및 생물학적 동등성 시험 가이드라인"을 소개하고자 한다., globunlin II의 두 component로 분리(分離)되었고 oryzenin은 시료(試料) 육우(陸羽) 132호(號), 등판(藤坂) 5호(號), 관산(關山) 재건(再建)에서는 oryzenin I, oryzenin II, oryzenin III의 3 component를 분리(分離)하였고 팔달(八達), 진흥(振興), 서광(瑞光), 은방주(銀坊主), 다마금(多摩錦), Pin Galw56의 6시료(試料)에서는 oryzenin II, oryzenin III의 2 component만 분리(分離)하였다. 3. 각(各) fraction을 여지전기영동(濾紙電氣泳動)하여 각(各) component의 농도곡선(濃度曲線)에 의해 함량(含量)을 구(求)한 결과(結果) albumin 0.26%, globulin I, 0.35%, globulin II, 0.32%, prolamin, 0.41%, oryzenin I, 0.30%, oryzenin II, 2.23%, oryzenin III, 2.66%이었다.미국의 경우 3일 이내 전화나 FAX로 보고하고, 10일이내 문서로써 보고하도록 되어 있다. 이러한 임상적 안전성 자료 관리에 대한 신속 보고 정의 및 기준에 대해서는 ICH에서도 논의가 되어 Step 4까지 다다르고 있으므로 앞으로 각 국에서의 적용만 남아있는 상태이고 보면 신약의 개발과 더불어 임상시험에서의 부작용등의 안전성 자료 관리에 대한 중요성은 더욱 강조될 것이다.공하는 음식의 섭취정도에도 영향을 주고 있으므로 학생들에게 학부모와 전담교사 및 학교영양사는 학생들에게 이상적인 아침식사에 대한 교육은 물론이고 아침식사를 실천할 수 있도록 다양한 방안에 대해 함께 연구해야 하겠다. 정부차원에서 학교급식에 아침식사 프로그램을 도입할

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The Effect of Garlic and Medicinal Plants Extracts on the Liver Function and Lipid Metabolism of Rats Administered with Alcohol (마늘과 한약재 추출물의 혼합급이가 알코올 투여에 의한 간기능 및 지질대사에 미치는 영향)

  • Lee, Soo-Jung;Kang, Min-Jung;Shin, Jung-Hye;Kim, Jeong-Gyun;Kang, Shin-Kwon;Sung, Nak-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.561-568
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    • 2009
  • To investigate the effects of garlic and medicinal plants extracts (GP) on liver function and lipid metabolism of rat administered with ethanol chronically, Sprague-Dawly male rats were fed with a basial diet (Normal), a basial diet plus ethanol (Control, 10 mL of 20% ethanol/kg b.w/day), a control diet plus 0.5% garlic and 1.0% medicinal plants extracts (GP-I), and a control diet plus 1.0% garlic and medicinal plants extracts (GP-II) for 4 weeks. Blood glucose in GP group was significantly decreased, but not significantly different between GP-I and GP-II group. Albumin content of serum was significantly increased in GP groups, while total lipid, cholesterol and triglyceride of serum were significantly decreased in GP group. Total cholesterol and triglyceride were not significantly different between GP-I and GP-II group. LDL-cholesterol in blood was decreased to 58% in GP-I group and 73% in GP-II group compared to the control group, it's contents were the lowest amounts among the normal, control and experimented groups. Lipid levels in liver of rat administered with alcohol were decreased in GP group and significantly different in GP-II group. GOT and r-GTP activities were significantly higher in control than normal group, while GPT and ALP activities were not significant in groups administered with alcohol. Activities of GOT, GPT and r-GTP were significantly lower in GP group than control group, while ALP activity was not significant in all groups. TBARS contents were not significant in serum, but it's contents in liver were significantly decreased in GP groups than control group. DPPH radical scavenging ability in serum and liver was significantly increased in GP groups. These results indicate that garlic and medicinal plants extracts were effective in improving and protecting liver disorder induced from long-term alcohol consumption.