• 제목/요약/키워드: cholesterol assay kit

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Culture Tube 방법에 의한 난황중의 Cholesterol 정량 (Culture Tube Method for the Determination of Total Cholesterol in Egg Yolk Lipid)

  • Ha, Yeong-Lae;Kim, Jeong-Ok
    • 한국식품영양과학회지
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    • 제23권6호
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    • pp.1032-1037
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    • 1994
  • A simple, reproducible , and accurate enzymatic method using a cholesterol assay kit was developed to quantify total cholesterol content in egg yolk. Total egg yolk lipid was extracted with hexane : isopropanol(3 : 2, v/v) mixture. Samples containing various amount of the total lipid(0-3mg) in optically identifical culture tubes were reacted for 10 min in a water bath (37$^{\circ}C$) with the enzyme solution (5ml) from the cholesterol assay kit. Cholesterol content of the reaction mixturesin culture tubes was spectrophotometrically determined by two different ways : (1) using the culture tube as a curvette(designate culture tube method ; CTM) and (2) the quartz cvette containing the reaction mixture transferred from the culture tube (designate standard cvette method, SCM). CTM revealed lower cholesterol content in 0.1-1.0mg lipid sample range that SCM did, but not significant. For more than 2.0mg lipid sample, CTM gave significantly (p<0.01) lower cholesterol content relative to that by SCM, suggesting that SCM give a false positive result from the sample containing more than 2 mg lipid due to the interference of absorbance by lipid dispersed in the reaction solution . Cholesterol content of less than 1.0mg lipid sample by CTM was proportional to the amount of lipid used, but its linear relationship was not seen in more than 2mg lipid sample. Thus, to determine the appropriate lipid amounts (mg) analyzed . A constant level (41$\mu\textrm{g}$/mg) of cholesterol concentration was observed from the sample containing 0.1-1mg lipid. after which the cholesterol level was dropped to less than 41$\mu\textrm{g}$ /mg. Cholesterol concentration in egg yolk samples quantified by CTM was in accordance with that by GC method. These results suggest that CTM is an useful method for the quantification of cholesterol in egg yolk lipid and other lipids as well.

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고콜레스테롤 식이 섭취 쥐에서 quercetin의 간 AMPK 및 microRNA-21 조절을 통한 지질대사 개선 효과 (Effects of quercetin on the improvement of lipid metabolism through regulating hepatic AMPK and microRNA-21 in high cholesterol diet-fed mice)

  • 이막순;김양하
    • Journal of Nutrition and Health
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    • 제55권1호
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    • pp.36-46
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    • 2022
  • Quercetin의 지질대사 개선 효과에 대한 작용기전을 확인하기 위해 C57BL/6J mouse를 사용하여 실험을 수행하였다. 고콜레스테롤혈증을 유도하기 위해 6주간 1% 콜레스테롤과 0.5% cholic acid를 함유하는 고콜레스테롤 식이를 급여하였으며, quercetin은 0.05%와 0.1%의 수준으로 고콜레스테롤 식이에 추가하여 같은 기간 동안 제공하였다. Quercetin은 혈청과 간의 중성지방 및 콜레스테롤 수준을 용량 의존적으로 감소하는 것으로 나타났다. 고콜레스테롤 식이를 섭취한 쥐의 간에서 지방 합성을 촉진하는 SREBP-1c, ACC1 및 FAS 유전자 발현이 quercetin 섭취에 의해 억제되는 것을 확인하였다. Quercetin은 간세포 내에서 에너지 대사를 조절하는 AMPK 활성을 증가시켰다. 이에 반해 암세포 증식을 촉진하고 지방간에서 높게 발현되는 miR-21 발현은 quercetin 섭취에 의해 감소되었다. 본 연구의 결과는 quercetin이 고콜레스테롤 식이 섭취 쥐에서 혈청과 간의 지질 수준을 낮추는 지질대사 개선 효과가 있으며, 이러한 효과의 일부는 간 내 지방합성 유전자 (SREBP-1c, ACC1 및 FAS) 발현, AMPK 활성 및 miR-21 조절을 통해 매개된다는 것을 시사한다.

지방간 유도 세포모델에서 황금(黃芩), 목향(木香), 시호(柴胡) 복합 추출물의 이상지질혈증에 대한 효과 (Anti-hyperlipidemic Effects of Scutellariae Radix, Aucklandiae Radix and Bupleuri Radix (SAB) extract in FL83B cells)

  • 도현주;김경호;오태우
    • 대한본초학회지
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    • 제35권5호
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    • pp.23-31
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    • 2020
  • Objectives : This study was conducted to evaluate the anti-hyperlipidemia effect of Scutellariae Radix, Aucklandiae Radix and Bupleuri Radix(SAB). Methods : FL83B cells were mouse liver hepatocytes, and we used this cell line. FL83B cells were treated with 0.5 mM oleic acid(OA) for 24 h, SAB extract was treated. After OA treatment, intracellular triglyceride (TG) and free fatty acid contents were measured with AdiopoRed™ assay and Free Fatty Acid Quantitation assay kit, respectively. Further, we evaluated several lipogenesis and metabolic markers such as sterol regulatory element-binding transcription factor-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), 3-hydroxy3-methyl-glutaryl CoA reductase (HMGCR), hormone-sensitive lipase (HSL), carnitine palmitoyltransferase (CPT-1), peroxisome proliferator activated receptor alpha (PPARα), and cluster of differentiation (CD36) using RT-PCR and Western-blot analysis. Results : OA markedly increased intracellular TG and free fatty acid, which plays a key role in reducing hepatic lipid accumulation, in FL83B cells. These increases were alleviated by SAB extract. The mRNA and protein expression of Fatty acid(FA) oxidation factors (CPT-1, PPARα), lipolysis factor(HSL), FA transporter(CD36), cholesterol synthesis factors (HMGCoA) and Lipodenesis (SREBP-1c, FAS, and ACC-1) were significantly increased by treatment of SAB extract in the OA-induced fatty liver cell model. Conclusions : In summary, the treat of SAB extract showed a significant reduction of the influx of fatty acids into hepatocytes, promoted the oxidation of fatty acids, and regulated fat synthesis-related factors, thereby regulating the accumulation of TG and free fatty acids.

Treadmill 운동과 DNA 및 칡 Catechin 섭취가 흰쥐 생체내 지방조성과 항산화 활성에 미치는 영향 (Effect of the Treadmill Exercise and the Intake of DNA and Crude Catechin (from Puerariae thunbergiana Roots) on the Body Fat Composition and the Antioxidant Activity in Rats)

  • 이치호;조진국;이은정;손영희;남혜영;최일신
    • 한국축산식품학회지
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    • 제23권2호
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    • pp.180-185
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    • 2003
  • 본 연구는 운동부하와 DNA 및 칡 crude catechin의 급여에 의한 생체내 성분 변화를 알아보기 위하여 흰쥐를 대조군, 운동부하군, 운동부하+DNA 및 운동부하+DNA+crude catechin 첨가군으로 나누어 사육하였다. 각각 0.4% DNA와 0.1% 칡crude catechins을 해당 횐쥐에 급식시켰으며, 운동부하군은 Treadmill에서 20 m/min의 속도로 6주 동안 운동시켰다. 사육실험종료 후 체중 증체량과 체지방무게를 먼저 칭량한 후, 항산화활성과 체지방조성을 kit assay를 이용하여 측정하였다. 6주후 각 군간의 최종 체중에는 유의적인 차이가 없었으나 간장, 부고환주위지방 및 신장주위지방은 운동부하군, 운동부하+DNA 첨가군 및 운동부하+DNA+crude catechin 첨가군이 대조군에 비해 유의하게 낮은 값(p<0.05)을 나타냈다. 또, 혈중 phospholipid, cholesterol 및 triglyceride농도는 운동부하군, 운동부하+DNA 첨가군 및 운동부하+DNA+crude catechin 첨가군이 대조군에 비해 유의하게 낮았으나(p<0.05), HDL-cholesterol은 유의하게 높은 값을 나타냈다. GPT 활성은 각 군간에 유의차가 없었으나, GOT활성은 운동부하를 처리한 군에서 적은 수치를 나타냈으며, TBA가는 각 군간에 유의 차가 없었다. 또, 혈중 NEFA 수준은 운동부하+DNA+crude catechin 첨가군이 대조군에 비해 유의하게 높은 값을 나타냈다.(p<0.05). 이상의 실험에서 운동과 더불어 crude catechin 섭취가 유의한 차이를 나타내지는 않았지만 혈청지질의 성분의 향상에 보조역할을 할 것으로 사료된다.

Effects of natural raw meal (NRM) on high-fat diet and dextran sulfate sodium (DSS)-induced ulcerative colitis in C57BL/6J mice

  • Shin, Sung-Ho;Song, Jia-Le;Park, Myoung-Gyu;Park, Mi-Hyun;Hwang, Sung-Joo;Park, Kun-Young
    • Nutrition Research and Practice
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    • 제9권6호
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    • pp.619-627
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    • 2015
  • BACKGROUND/OBJECTIVES: Colitis is a serious health problem, and chronic obesity is associated with the progression of colitis. The aim of this study was to determine the effects of natural raw meal (NRM) on high-fat diet (HFD, 45%) and dextran sulfate sodium (DSS, 2% w/v)-induced colitis in C57BL/6J mice. MATERIALS/METHODS: Body weight, colon length, and colon weight-to-length ratio, were measured directly. Serum levels of obesity-related biomarkers, triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), insulin, leptin, and adiponectin were determined using commercial kits. Serum levels of pro-inflammatory cytokines including tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, and IL-6 were detected using a commercial ELISA kit. Histological study was performed using a hematoxylin and eosin (H&E) staining assay. Colonic mRNA expressions of TNF-${\alpha}$, IL-$1{\beta}$, IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were determined by RT-PCR assay. RESULTS: Body weight and obesity-related biomarkers (TG, TC, LDL, HDL, insulin, leptin, and adiponectin) were regulated and obesity was prevented in NRM treated mice. NRM significantly suppressed colon shortening and reduced colon weight-to-length ratio in HFD+DSS induced colitis in C57BL/6J mice (P < 0.05). Histological observations suggested that NRM reduced edema, mucosal damage, and the loss of crypts induced by HFD and DSS. In addition, NRM decreased the serum levels of pro-inflammatory cytokines, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 and inhibited the mRNA expressions of these cytokines, and iNOS and COX-2 in colon mucosa (P < 0.05). CONCLUSION: The results suggest that NRM has an anti-inflammatory effect against HFD and DSS-induced colitis in mice, and that these effects are due to the amelioration of HFD and/or DSS-induced inflammatory reactions.