• 제목/요약/키워드: HMG-1

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Development of Marker-free Transgenic Rice for Increasing Bread-making Quality using Wheat High Molecular Weight Glutenin Subunits (HMW-GS) Gene (밀 고분자 글루테닌 유전자를 이용하여 빵 가공적성 증진을 위한 마커 프리 형질전환 벼의 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1317-1324
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    • 2013
  • High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant $T_0$ plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice $T_1$ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the $T_1$ generation.

Development of Marker-free Transgenic Rice Expressing the Wheat Storage Protein, Glu-1Dy10, for Increasing Quality Processing of Bread and Noodles (빵과 면의 가공적성 증진을 위한 밀 저장단백질 Glu-1Dy10을 발현하는 마커프리 형질전환 벼 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Hur, Yeon-Jae;Kim, Tae-Heon;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Lee, Seung-Sik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.24 no.6
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    • pp.618-625
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    • 2014
  • Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.

GnRH Agonist Stimulation Test (GAST) for Prediction of Ovarian Response in Controlled Ovarian Stimulation (COH) (난소기능평가를 위한 Gonadotropin Releasing Hormone Agonist Stimulation Test (GAST)의 효용성에 관한 연구)

  • Kim, Mee-Ran;Song, In-Ok;Yeon, Hye-Jeong;Choi, Bum-Chae;Paik, Eun-Chan;Koong, Mi-Kyoung;Song, Il-Pyo;Lee, Jin-Woo;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.163-170
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    • 1999
  • Objectives: The aims of this study are 1) to determine if GAST is a better indicator in predicting ovarian response to COH compared with patient's age or basal FSH level and 2) to evaluate its role in detecting abnormal ovarian response. Design: Prospective study in 118 patients undergoing IVF-ET using GnRH-a short protocol during May-September 1995. Materials and Methods: After blood sampling for basal FSH and estradiol $(E_2)$ on cycle day two, 0.5ml (0.525mg) GnRH agonist ($Suprefact^{(r)}$, Hoechst) was injected subcutaneously. Serum $E_2$ was measured 24 hours later. Initial $E_2$ difference $({\Delta}E_2)$ was defined as the change in $E_2$ on day 3 over the baseline day 2 value. Sixteen patients with ovarian cyst or single ovary or incorrect blood collection time were excluded from the analysis. The patients were divided into three groups by ${\Delta}E_2$; group A (n=30):${\Delta}E_2$<40 pg/ml, group B (n=52): 40 pg/ml${\leq}{\Delta}E_2$<100 pg/ml, group C (n=20): ${\Delta}E_2{\leq}100$ pg/ml. COH was done by GnRH agonist/HMG/hCG and IVF-ET was followed. Ratio of $E_2$ on day of hCG injection over the number of ampules of gonadotropins used ($E_2hCGday$/Amp) was regarded as ovarian responsiveness. Poor ovarian response and overstimulation were defined as $E_2$ hCGday less than 600 pg/ml and greater than 5000 pg/ml, respectively. Results: Mean age $({\pm}SEM)$ in group A, B and C were $33.7{\pm}0.8^*,\;31.5{\pm}0.6\;and\;30.6{\pm}0.5^*$, respectively ($^*$: p<0.05). Mean basal FSH level of group $A(11.1{\pm}1.1mlU/ml)$ was significantly higher than those of $B(7.4{\pm}0.2mIU/ml)$ and C $(6.8{\pm}0.4mIU/ml)$ (p<0.001). Mean $E_2hCGday$ of group A was significantly lower than those of group B or C, i.e., $1402.1{\pm}187.7pg/ml,\;3153.2{\pm}240.0pg/ml,\;4078.8{\pm}306.4pg/ml$ respectively (p<0.0001). The number of ampules of gonadotropins used in group A was significantly greater than those in group B or C: $38.6{\pm}2.3,\;24.2{\pm}1.1\;and\;18.5{\pm}1.0$ (p<0.0001). The number of oocytes retrieved in group A was significantly smaller than those in group B or C: $6.4{\pm}1.1,\;15.5{\pm}1.1\;and\;18.6{\pm}1.6$, respectively (p<0.0001). By stepwise multiple regression, only ${\Delta}E_2$ showed a significant correlation (r=0.68, p<0.0001) with $E_2HCGday$/Amp, while age or basal FSH level were not significant. Likewise, only ${\Delta}E_2$ correlated significantly with the number of oocytes retrieved (r=0.57, p<0.001). All four patients whose COH was canceled due to poor ovarian response belonged to group A only (Fisher's exact test, p<0.01). Whereas none of 30 patients in group A (0%) had overstimulation, 14 patients among 72 patients (19.4%) in group B and C had overstimulation (Fisher's exact test, p<0.01). Conclusions: These data suggest that initial $E_2$ difference after GAST may be a better prognostic indicator of ovarian response to COH than age or basal FSH level. Since initial $E_2$ difference demonstrates significant association with abnormal ovarian response such as poor ovarian response necessitating cycle cancellation or overstimulation, GAST may be helpful in monitoring and consultation of patients during COH in IVF-ET cycle.

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Effects of Coenzyme Q10 on the Expression of Genes involved in Lipid Metabolism in Laying Hens (Coenzyme Q10 첨가 급여가 산란계의 지방대사 연관 유전자 발현에 미치는 영향)

  • Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.43 no.1
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    • pp.47-54
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    • 2016
  • The aim of this study was to investigate the expression patterns of key genes involved in lipid metabolism in response to dietary Coenzyme Q10 (CoQ10) in hens. A total of 36 forty week-old Lohmann Brown were randomly allocated into 3 groups consisting of 4 replicates of 3 birds. Laying hens were subjected to one of following treatments: Control (BD, basal diet), T1 (BD+ CoQ10 100 mg/kg diet) and T2 (BD+ micellar of CoQ10 100 mg/kg diet). Birds were fed ad libitum a basal diet or the basal diet supplemented with CoQ10 for 5 weeks. Total RNA was extracted from the liver for quantitative RT-PCR. The mRNA levels of HMG-CoA reductase(HMGCR) and sterol regulatory element-binding proteins(SREBP)2 were decreased more than 30~50% in the liver of birds fed a basal diet supplemented with CoQ10 (p<0.05). These findings suggest that dietary CoQ10 can reduce cholesterol levels by the suppression of the hepatic HMGCR and SREBP2 genes. The gene expressions of liver X receptor (LXR) and SREBP1 were down regulated due to the addition of CoQ10 to the feed (p<0.05). The homeostasis of cholesterol can be regulated by LXR and SREBP1 in cholesterol-low-conditions. The supplement of CoQ10 caused a decreased expression of lipid metabolism-related genes including $PPAR{\gamma}$, XBP1, FASN, and GLUTs in the liver of birds (p<0.05). These data suggest that CoQ10 might be used as a dietary supplement to reduce cholesterol levels and to regulate lipid homeostasis in laying hens.

Inherited metabolic diseases in the urine organic acid analysis of complex febrile seizure patients (복합 열성경련 환자의 소변 유기산 분석에서 나타난 유전대사질환)

  • Cheong, Hee Jeong;Kim, Hye Rim;Lee, Seong Soo;Bae, Eun Joo;Park, Won Il;Lee, Hong Jin;Choi, Hui Chul
    • Clinical and Experimental Pediatrics
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    • v.52 no.2
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    • pp.199-204
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    • 2009
  • Purpose : Seizure associated with fever may indicate the presence of underlying inherited metabolic diseases. The present study was performed to investigate the presence of underlying metabolic diseases in patients with complex febrile seizures, using analyses of urine organic acids. Method : We retrospectively analyzed and compared the results of urine organic acid analysis with routine laboratory findings in 278 patients referred for complex febrile seizure. Results : Of 278 patients, 132 had no abnormal laboratory findings, and 146 patients had at least one of the following abnormal laboratory findings: acidosis (n=58), hyperammonemia (n=55), hypoglycemia (n=21), ketosis (n=12). Twenty-six (19.7 %) of the 132 patients with no abnormal findings and 104 (71.2%) of the 146 patients with statistically significant abnormalities showed abnormalities on the organic acid analysis (P<0.05). Mitochondrial respiratory chain disorders (n=23) were the most common diseases found in the normal routine laboratory group, followed by PDH deficiency (n=2) and ketolytic defect (n=1). In the abnormal routine laboratory group, mitochondrial respiratory chain disorder (n=29) was the most common disease, followed by ketolytic defects (n=27), PDH deficiency (n=9), glutaric aciduria type II (n=9), 3-methylglutaconic aciduria type III (n=6), biotinidase deficiency (n=5), propionic acidemia (n=4), methylmalonic acidemia (n=2), 3-hydroxyisobutyric aciduria (n=2), orotic aciduria (n=2), fatty acid oxidation disorders (n=2), 2-methylbranched chain acyl CoA dehydrogenase deficiency (n=2), 3-methylglutaconic aciduria type I (n=1), maple syrup urine disease (n=1), isovaleric acidemia (n=1), HMG-CoA lyase deficiency (n=1), L-2-hydroxyglutaric aciduria (n=1), and pyruvate carboxylase deficiency (n=1). Conclusion : These findings suggest that urine organic acid analysis should be performed in all patients with complex febrile seizure and other risk factors for early detection of inherited metabolic diseases.

Effect of Mulberry (Morus alba L.) Extract on Blood Flow Improvement (오디 추출물(Morus alba L.)의 혈행개선 효과)

  • Park, Youn-Sil;Kang, Seong-Sun;Choi, Hyoung-Ja;Yang, Sung-Jun;Shon, Ho-Hyeong;Seo, Hyeong-Ho;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.498-506
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    • 2014
  • The objective of this study was to investigate the beneficial effects of mulberry extract (MBE) on blood flow improvement. The $SC_{50}$ value for the DPPH radical scavenging activity of MBE was $89.36{\pm}5.46{\mu}g/mL$. Analysis of the cellular toxicity of MBE on RAW 264.7 and HepG2 cells showed no toxicity under a concentration of 2,500 ${\mu}g/mL$. We found that MBE inhibited the enzyme activity of cyclooxygenase (COX)-2 as well as oxidation of human LDL. Western blotting analysis showed that MBE inhibited protein expression of COX-2 and 5-lipoxygenase in RAW 264.7 cells. In addition, MBE inhibited protein expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in human umbilical vein endothelial cells. Furthermore, MBE reduced the serum levels of total cholesterol and C-reactive protein in a concentration-dependent manner. These results both in vitro and in vivo suggest that MBE can be employed for the improvement of blood flow.

In Vitro Maturation of Tiger Oocytes : A Case Report (호랑이 난자의 체외성숙)

  • Lee, H.-S.;Yin, X.-J.;Lee, Y.-H.;Min, W.-K.;Kim, T.-S.;Choi, J.-W.;Yoon, B.-C.;Kim, J.-I.;Kong, I.-K.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.185-189
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    • 2004
  • The purpose of this study was to determine the possibility of in vitro maturation of tiger oocytes. Immature oocytes were recovered from a pair of ovaries. A total of 78 oocytes was collected, of which forty three were classified as good oocytes with compact cumulus cells and uniform cytoplasm. Forty three COCs were in vitro matured at $39^{\circ}C$, 5% CO2 in air atmosphere for 48 h in a IVM medium (TCM-199 supplement with 10% FBS, 0.6 mM cysteine, 0.2 mM pyruvic acid and 10 IU/mL HMG). Experiment I: the morphologic evaluation was conducted by measuring the diameter of oocytes with or without ZP, the thickness of ZP and the diameter of cytoplasm by microeyepiece at the same magnification (${\times}$100). Experiment II: the evaluation of meiotic development was conducted of the nuclear development stage of tiger oocytes. The results were summarized as follows: 1. The diameter of tiger oocytes $(176.5\pm6.1{\mu}m)$ with ZP was significantly (p<0.05) bigger than that of bovine oocytes $(150.7\pm4.9{\mu}m).$ The ZP thickness of tiger oocytes $(20.4\pm2.9{\mu}m)$ was significantly (p<0.05) bigger than that of bovine oocytes $(12.0\pm2.6{\mu}m;$ p<0.05). However, there was no significant difference in the diameter of cytoplasm (without ZP) between tiger $(122.1\pm9.7{\mu}m)$ and bovine oocytes $(118.7\pm7.5{\mu}m).$ 2. The rates of meiotic development of tiger oocytes were achieved GV (12.5 %) and MII (50.0%), respectively. These results indicated that tiger oocytes could be developed to MII in in vitro culture system.

Anticancer (in vitro) and Antiallergy Effects of Rice Bran Extracts (쌀겨 추출물의 항암효과(in vitro) 및 항알레르기 효과)

  • Choi, Hyun-Im;Ye, Eun-Ju;Kim, Soo-Jung;Bae, Man-Jong;Yee, Sung-Tae;Park, Eun-Jung;Park, Eun-Mi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1297-1303
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    • 2006
  • This study was conducted to investigate the anticancer (in vitro) and antiallergy effects of rice bran extracts. In an anticancer test using Hep3B cells and HeLa cells, water and 60% ethanol extracts of rice bran inhibited the growth of Hep3B and HeLa cell lines and morphological changes were also observed. In Hep3B cell lines, water extract of rice bran showed a higer antiproliferating effect than 60% ethanol extract. The growth-inhibitory effect against HeLa cells were 30.9% for $1,000{\mu}g/mL$, 88.8% for $3,000{\mu}g/mL$ rice bran water extract. The expressions of $Fc{\varepsilon}RI$ mRNA and c-kit in HMC-1 (human mast cell) were decreased by 60% ethanol treatment but tryptase mRNA was not changed. The extracts of rice bran inhibited histamine release from RPMC (rat peritoneal mast cell) activated by compound 48/80. Rice bran water extract showed inhibitory effect of 87% at $0.01{\mu}g/mL$ concentration and 60% ethanol extract inhibited the release of histamine by 86% at $100{\mu}g/mL$ concentration.

Prognostic Value of Day 3 Inhibin-B on Assisted Reproductive Technology Outcome (보조 생식술 결과에 있어서 기저혈중 Inhibin-B의 예후인자로서의 유용성)

  • Bai, Sang-Wook;Kim, Jin-Young;Lee, Kyung-Sool;Won, Jong-Gun;Lee, Yong-Joo;Yi, Ji-Won;Chang, Kyung-Hwan;Lee, Byung-Seok;Park, Ki-Hyun;Cho, Dong-Jae;Song, Chan-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.217-223
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    • 1997
  • This study was performed to determine if women with day 3 serum inhibin-B concentrations <45pg/ml (conversion factor to SI unit, 1.00) demonstrate a poor response to ovulation induction and assisted reproductive technology outcome to women with inhibin-B${\ge}45pg$/ml, independant of day 3 FSH, E2 and patient age. From Jan 1996 to Dec 1996, 16 volunteers patients who underwent 25 IVF cycles with luteal phase GnRH agonist suppression and HMG stimulation were allocated to the study group. We evaluated day 3 serum inhibin-B, FSH, E2, peak E2, cancellation rate per initiated cycle (%) and clinical pregnancy rate per initiated cycle (%) according to the above two groups and independent of patient age, day 3 FSH, day 3 E2 and all of above combined. Women with day 3 serum inhibin-B${\ge}45pg$/ml demonstrated higher average day 3 inhibin-B level, clinical pregnancy rate per initiated cycle ($20.3{\pm}2.5$ pg/ml vs $80.9{\pm}5.0$ pg/ml, p<0.05; 24.8% vs 8.5%, p<0.05) and lower day 3 FSH level, cancellation rate per initiated cycle ($6.9{\pm}0.3$ mIU/ml vs $8.5{\pm}0.5$ mIU/ml, p<0.05; 1.5% vs 9.0%, p<0.05). Women with day 3 serum inhibin${\ge}45pg$/ml and age<40 year demonstrated higher pregnancy rate per initiated cycle (28.2% vs 7.4%, p<0.05) and lows. day 3 FSH level, cancellation rate per initiated cycle ($6.9{\pm}0.5$ mIU/ml vs $8.2{\pm}0.7$ mIU/ml, p<0.05; 1.0% vs 9.0%, p<0.05). Women with day 3 serum inhibin${\ge}45pg$/ml and day 3 FSH<15mIU/ml demonstrated higher pregnancy rate per initiated cycle (33.5% vs 9.5%, p<0.05) and lower day 3 FSH level, cancellation rate per initiated cycle ($7.7{\pm}0.2$ mIU/ml vs $8.5{\pm}0.5$ mIU/ml, p<0.05; 1.5% vs 10.0%, p<0.05). Women with day 3 serum inhibin${\ge}45pg$/ml and day 3 E2<50pg/ml demonstrated higher pregnancy rate per initiated cycle (30.0% vs 9.5%, p<0.05) and lower cancellation rate per initiated cycle (1.5% vs 9.5%, p<0.05). Women with day 3 serum inhibin${\ge}45pg$/ml, age<40 year, day 3 FSH<15mIU/ml and day 3 E2<50pg/ml demonstrated higher pregnancy rate per initiated cycle (30.0% vs 10.8%, p<0.05) and lower day 3 FSH level, cancellation rate per initiated cycle ($6.8{\pm}0.6$ mIU/ml vs $8.4{\pm}0.9$ mIU/ml, p<0.05; 1.5% vs 7.8%, p<0.05). Therefore women with low day 3 serum inhibin-B concentrations demonstrate a poorer response to ovulation induction and are less likely to conceive a clinical pregnancy though ART relative to women with high day 3 inhibin-B and day 3 serum inhibin-B, in addition to a day 3 FSH, E2 and patient age, appears helpful in prediction in IVF-ET outcome.

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Comparison of the Effects of Cyclodextrin-Naringin Inclusion Complex with Naringin on Lipid Metabolism in Mice Fed a High-Fat Diet (고지방식이를 섭취한 마우스에서 나린진과 나린진-사이클로텍스트린 포접화합물의 지질대사에 대한 영향 비교)

  • Jeon, Seon-Min;Choi, Myung-Sook
    • Journal of the East Asian Society of Dietary Life
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    • v.20 no.1
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    • pp.20-29
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    • 2010
  • Naringin has antioxidant and antihyperlipidemic properties, however, phenolic compounds including naringin are unstable in the presence of light, heat and oxygen. Beta-cyclodextrin ($\beta$-CD) is a cyclic heptamer composed of seven glucose units that enhances the stability and solubility of molecules through the formation of inclusion complexes. This study was conducted out to compare the effects of CD-naringin (CD-N) inclusion complexes with naringin on lipid metabolism in high fat-fed animals. Male C57BL/6 mice were fed either CD-N (0.048%, w/w) or naringin (N, 0.02%, w/w) in a 20% high-fat (HFC, 15% lard, 5% corn oil, w/w) diet for 10 weeks. Orlistat (Xenical, 0.01%, w/w) was used as a positive control (PC). There were no differences in body weight, food intake, liver and heart weights, plasma triglyceride(TG), leptin, adiponectin, resistin, IL-$1{\beta}$ and IL-6 concentrations, and hepatic $\beta$-oxidation, carnitine palmitoyl transferase(CPT), glucose-6-phosphate dehydrogenase (G6PD) and malic enzyme activities between the HFC and CD-N groups or between the HFC and N groups. However, both CD-naringin and naringin supplementation les to a significant reduction in the epididymal and perirenal white adipose tissue weights, plasma free fatty acid, insulin and blood glucose concentrations, hepatic cholesterol and TG contents and hepatic fatty acid synthase (FAS), phosphatidate phosphohydrolase (PAP) and HMG-CoA reductase activities compared to the HFC group. The plasma HDL-cholesterol concentration was significantly higher in CD-N and N groups than in HF and PC groups. These results indicate that both CD-naringin and naringin supplementation effectively improved plasma and hepatic lipid metabolism without differences between CD-N and naringin groups.