• 제목/요약/키워드: citrate synthase

검색결과 66건 처리시간 0.023초

유산균을 활용한 미역발효추출물이 3T3-L1에서 지방세포 분화에 미치는 영향 (Effects of Undaria pinnatifida and Leuconostoc mesenteroides Fermented Extracts on Adipocyte Differentiation in 3T3-L1 Cells)

  • 문민우;라채훈
    • 한국미생물·생명공학회지
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    • 제52권3호
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    • pp.298-303
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    • 2024
  • 유산균 L. mesenteroides SGL152를 이용한 미역발효 추출물로부터 3T3-L1 지방세포 분화에 미치는 영향을 살펴보기 위해 MTT assay, Oil Red O staining, RT-qPCR 실험을 진행하였다. 유산균의 미역발효 추출물(UFM)의 농도별 세포 독성 실험 결과, 200 ㎍/ml 농도 이하에서는 비세포독성으로 판단된다. 또한 3T3-L1 지방세포 분화와 동시에 추출물을 농도별로 처리하여 분화를 진행한 결과, 대조군과 비교하여 유의적으로 지방구 형성 및 지방분화를 억제하였다. 이러한 연구결과는 유산균을 이용한 해조류 발효과정 및 복합추출물에 있어서 유용한 정보를 제공할 것으로 판단된다.

Nutritional Regulation of GLUT Expression, Glucose Metabolism, and Intramuscular Fat Content in Porcine Muscle

  • Katsumata, M.;Kaji, Y.;Takada, R.;Dauncey, M.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권8호
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    • pp.1297-1304
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    • 2007
  • We conducted a series of investigations in order to elucidate role of nutritional status in regulating GLUT expression and energy metabolism in porcine muscle. Firstly, the role of mild undernutrition in regulating muscle GLUT gene expression and function was studied in growing pigs (3 wk of age) on a high (H) or low (L) food intake (H = 2L) at $35^{\circ}C$ or $26^{\circ}C$. Low food intake selectively upregulates GLUT1 and GLUT4 gene expression; mRNA levels were elevated in longissimus dorsi (L. dorsi) and rhomboideus muscles but not in diaphragm or cardiac muscles. Our next step was to determine whether dietary lysine, a major primary limiting amino acid in diets for pigs, affects muscle GLUT4 expression. Pigs of 6 wk of age were pair-fed a control or low lysine (LL) diet. The control diet contained optimal amounts of all essential amino acids, including 1.15% lysine. The LL diet was similar but contained only 0.70% lysine. GLUT4 mRNA expression was upregulated by the LL diet in L. dorsi and rhomboideus muscles, whereas that in cardiac muscle was unaffected. GLUT4 protein abundance was also higher in rhomboideus muscle of animals on the LL diet. We conducted another investigation in order to elucidate effects of the LL diet on post-GLUT4 glucose metabolism. Activity of hexokinase was unaffected by dietary lysine levels while that of citrate synthase was higher both in L. dorsi and rhomboideus muscles of pigs fed on the LL diet. Glucose 6-phosphate content was higher in L. dorsi msucle in the LL group. Glycogen content was higher both in L. dorsi and rhomboideus muscles in the LL group. Further, we determined the effects of dietary lysine levels on accumulation of intramuscular fat (IMF) in L. dorsi muscle of finishing pigs. A low lysine diet (lysine content was 0.40%) meeting approximately 70% of the requirement of lysine was given to finishing pigs for two months. IMF contents in L. dorsi of the pigs given the low lysine diet were twice higher than those of the pigs fed on a control diet (lysine content was 0.65%). Finally, we proved that a well known effect of breadcrumbs feeding to enhance IMF of finishing pigs could be attributed to shortage of amino acids in diets including breadcrumbs.

홍경천(紅景天)추출물(KH101)이 강제유영 흰쥐의 피로회복에 미치는 영향 (Effects of Rhodiola rosea (KH101) on Anti-fatigue in Forced Swimming Rats)

  • 정혁상;김은영;심은섭;이현삼;문은정;김진화;김선여;손영주;손낙원
    • 대한한방내과학회지
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    • 제29권4호
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    • pp.922-938
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    • 2008
  • Objectives : Rhodiola rosea has been used in herbal medicine to treat various conditions, such as antimelancholia, antifatigue, improvement of work competence and prevention of altitude sickness. In this study, we investigated effects of Rhodiola rosea extract (KH101) on fatigue in forced swimming rats. Methods : Sprague-Dawley rats were induced with fatigue by forced swimming, then rats in each group were treated with KH101. We observed changes of glucose, LDH and cortisol in serum and LDH, glycogen, hexokinase, citrate synthase MDH, SDH and CK in muscle. Results : Obtained results were as follows: 1. Continuance times of exercise significantly increased in all groups at day 1, in the 50 mg/kg concentration group at day 2, in all groups at day 3 and in the 50 mg/kg conc. group at day 4. 2. In serum, glucose significantly decreased in all concentration groups. 3. In the soleus muscle, LDH significantly decreased in the 50 mg/kg concentration group. HK significantly decreased in the 100 mg/kg conc. group. SDH significantly increased in the 100 mg/kg conc. MDH were significantly decreased in all conc. groups. 4. In the gastrocnemius muscle, HK significantly decreased in all concentration groups, while MDH significantly increased all conc. groups. Conclusions : It is concluded that the KH101 has and anti-fatigue effect in rats. Additional studies are needed to find the mechanism of the association between each single herb.

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복합 운동 훈련이 인간 골격근의 형태학적 변화와 유산소 대사관련 효소 활성에 미치는 영향 (Effect of combined endurance and weight training on muscle morphology and oxidative enzyme activities in human skeletal muscle)

  • 이길우;이종삼;이장규
    • 한국산학기술학회논문지
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    • 제15권1호
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    • pp.367-376
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    • 2014
  • 이 연구의 목적은 건강한 남자 14명을 대상자로 단기간의 지구성 훈련과 지구성 운동에 웨이트 운동을 병행한 복합훈련이 골격근 섬유의 형태 및 골격근 내 산화적 효소의 활성도에 미치는 영향을 구명하고자 하였다. 지구성 트레이닝 집단은 최대심박수의 70~80% 에서 40분간 자유형으로 수영을 6주간 실시하였으며 복합 트레이닝 집단은 동일한 수영을 실시한 후 웨이트 트레이닝을 추가로 실시하였다. 근섬유 횡단면적에 있어 복합훈련 집단과 지구성 훈련 집단 간에 차이가 나타나지 않았으며 훈련기간에 따라 복합훈련 집단에서 지구성 훈련 집단보다 증가한 것으로 나타났다. 모세혈관 수에 있어서도 집단 간 차이는 나타나지 않았으며 훈련기간에 따라 복합훈련 집단에서 지구성 훈련 집단보다 증가한 것으로 나타났다. 유산소성 대사 효소 중 CS, ${\beta}$-HAD 그리고 CPT의 활성도는 훈련형태에 따른 차이는 통계적으로 나타나지 않았으나 복합훈련 집단에서 지구성 훈련 집단에서 보다 더 큰 증가의 폭을 나타냈다. 이와 같은 결과를 종합해 볼 때 유산소성 훈련만을 실시할 때 보다 유 무산소 운동이 복합적으로 구성되어 있는 훈련의 수행이 근섬유의 에너지 대사 관련 효소들의 활성도 증가와 근 세포 모세혈관의 발달에 보다 효과적이라는 결론을 도출할 수 있었다.

The Effects of A High-Fat Diet on Pro- and Macro-Glycogen Accumulation and Mobilization During Exercise in Different Muscle Fiber Types and Tissues in Rats

  • Lee Jong-Sam;Eo Su-Ju;Cho In-Ho;Pyo Jae-Hwan;Kim Hyo-Sik;Lee Jang-Kyu;Kwon Young-Woo;Kim Chang-Keun
    • Nutritional Sciences
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    • 제8권3호
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    • pp.181-188
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    • 2005
  • We investigated the effects of diet manipulation on pro- and macro-glycogen accumulation and mobilization during exercise in different kinds of muscle fiber and tissue. Thirty-two Sprague-Dawley rats were divided into groups representing one of two dietary conditions: high fat (HF, n=16) or standard chow (CHOW, n=16). Each dietary group was fm1her divided into control (REST, n=8) and exercise (EXE, n=8). After an eight-week dietary intervention period, the animals in EXE swam for 3 hours while the animals in REST remained at rest Skeletal muscle (soleus, red gastrocnemius and white gastrocnemius) and liver samples were then dissected out and used for analyses. 1here was no statistical difference in body weight between the animals in the HF and mow groups (p>.05). Three hours of exercise significantly increased plasma free fatty acid (FFA) concentration in the animals in the CHOW group but not in the animals in the HF group. Both citrate. synthase (CS) and $\beta$-hydroxyacyl dehydrogenase ($\beta$-HAD) activities in skeletal muscles were higher in the HF group than in the mow group. CS and $\beta$-HAD activities were also the highest in red gastrocnemius and the lowest in white gastrocnemius. At both time points (i.e., rest and immediately after exercise) intramuscular triglyceride (IMTG) and liver TG concentrations were significantly higher in the HF compared to the CHOW. IMTG and liver TG changed selectively in the CHOW. Except in white gastrocnemius muscle, there was no significant difference in total glycogen content between HF and mow at rest. Although exercise significantly lowered total glycogen content in all groups and tissues (p<.05), the degree of reduction was markedly greater in the mow than in the HF. Whereas changes in proglycogen concentration showed a trend similar to those of total glycogen, alterations in macroglycogen concentrations clearly differed from those of total glycogen. Specifically, the degree of reduction of macroglycogen following three hours of exercise was substantially greater in the CHOW than in the HF. These results suggest that metabolic alterations induced by a long-term high fat diet may be caused by macro-glycogen rather than pro-glycogen.

쓴메밀 새싹 추출물의 히스톤 아세틸화 효소 활성 저해에 의한 비알코올성 지방간 억제 효능 (Effect of Tartary Buckwheat Sprout on Non-Alcoholic Fatty Liver Disease through Anti-Histone Acetyltransferase Activity)

  • 황진택;남태규;정민유;박재호;최효경
    • 한국식품영양과학회지
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    • 제46권2호
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    • pp.169-176
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    • 2017
  • 본 연구에서는 쓴메밀 새싹 추출물(TBS)을 대상으로 histone acetyltransferase(HAT) 활성 저해능을 평가하고 oleic acid와 palmitic acid(OPA)를 이용하여 HepG2 세포에서 비알코올성 지방간을 유도하여 그 효과를 검토하였다. HeLa 세포의 nuclear extract(NE)를 HAT의 source로 하여 in vitro에서 TBS에 의한 HAT 활성 저해능을 평가한 결과 추출물의 처리에 의하여 HAT 활성이 억제됨을 관찰할 수 있었다. 또한, 대표적인 HAT 단백질인 p300과 CBP를 이용하여 동일한 방식으로 HAT 억제능을 평가한 결과 TBS 처리에 의하여 두 단백질 모두 활성이 감소하였으며, 특히 TBS는 p300의 활성을 특이적으로 저해함을 확인할 수 있었다. 그뿐만 아니라 HepG2 세포에 $400{\mu}M$의 oleic acid 및 $100{\mu}M$의 palmitic acid와 함께 $200{\mu}g/mL$, $500{\mu}g/mL$의 TBS를 처리한 후 NE를 이용하여 세포 내 HAT 활성을 측정한 결과 역시 추출물 처리에 의하여 세포 내 HAT 활성이 저해되어 있음이 관찰되었다. TBS에 의한 HAT 활성의 억제는 세포 내 다양한 단백질들의 아세틸화 저해와 지질축적에 의하여 아세틸화 변형을 일으키는 것으로 알려진 histone H3K9, H4K8의 아세틸화 및 H3K36의 아세틸화를 감소시켰으며, 세포 내 지질합성과 관련된 대표적 유전자인 SREBP1c, ACLY, FAS의 전사 활성 역시 저해함을 관찰하였다. 이와 같은 변화를 통하여 OPA에 의하여 HepG2 세포 내에 축적되었던 지질은 TBS의 처리에 의하여 효과적으로 감소하였으며 이때 처리된 OPA와 소재에 의한 세포 내 독성은 관찰되지 않았다. 그러므로 이러한 결과는 TBS에 의한 HAT 활성의 저해가 히스톤 단백질의 아세틸활 변형을 억제하고 이를 통하여 지방 합성 관련 유전자들의 전사 활성을 감소시켜 결과적으로 세포 내 지질축적을 방지하는 것으로 생각되며, TBS은 비알코올성 지방간질환의 예방에 좋은 천연물 소재로 활용될 수 있을 것이라 여겨진다.

태충·삼음교의 침 자극이 Streptozotocin으로 유발된 당뇨쥐의 신장 손상에 미치는 영향 (The Effects of LR3 and SP6 Acupuncture on Renal Damage in Streptozotocin-induced Diabetic Mice)

  • 이초인;이현종;이윤규;임성철;김재수
    • Journal of Acupuncture Research
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    • 제32권3호
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    • pp.41-51
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    • 2015
  • Objectives : This study was performed to investigate the effects of $LR_3$ and $SP_6$ acupuncture on renal damage in streptozotocin(STZ)-induced diabetic mice. Methods : ICR male mice were stabilized for a week and divided into four groups: a normal mice group(N), no-acupuncture diabetic mice group(Control), $LR_3$ acupuncture diabetic mice group($LR_3$), and $SP_6$ acupuncture diabetic mice group($SP_6$). Diabetes was experimentally induced by intraperitoneal injection of STZ(150 mg/kg) in citrate buffer(pH 4.5). For two weeks, $LR_3$ and $SP_6$ acupunctures were administered bilaterally at each point once a day. After two weeks, the animals' weight was measured and they underwent a laparotomy. Serum glucose and blood urea nitrogen(BUN) were measured from the blood taken from the heart. We measured glucose, reactive oxygen species(ROS), peroxynitrite($ONOO^-$) and thiobarbituric acid reactive substances(TBARS) in the kidney and compared expression levels of superoxide dismutases(SOD), glutathione peroxidase(GPx), nuclear factor-kappa B(NF-${\kappa}B$), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase(iNOS) and Interleukin-1 beta(IL-$1{\beta}$). Results : BUN significantly decreased in $LR_3$, $SP_6$ compared to the control group. $LR_3$ showed significantly decreased glucose compared to the control group. $LR_3$, $SP_6$ significantly decreased in ROS and $ONOO^-$ compared to the control group. $LR_3$ significantly decreased in TBARS compared to the control group. $SP_6$ significantly increased in expressions of SOD-1, catalase, and GPx compared to the control group. $LR_3$, $SP_6$ significantly decreased in COX-2 compared to the control group. $SP_6$ significantly decreased in IL-$1{\beta}$ compared to the control group. Conclusions : This study suggests that $LR_3$ acupuncture may be effective in controlling glucose and lipid peroxidation and that $SP_6$ acupuncture may have anti-oxidative and anti-inflammatory effects on renal damage in STZ-induced diabetic mice.

Functional Characterization of the ${\alpha}$- and ${\beta}$-Subunits of a Group II Chaperonin from Aeropyrum pernix K1

  • Lee, Jin-Woo;Kim, Se Won;Kim, Jeong-Hwan;Jeon, Sung-Jong;Kwon, Hyun-Ju;Kim, Byung-Woo;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • 제23권6호
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    • pp.818-825
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    • 2013
  • We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.

Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권4호
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Comprehensive investigations of key mitochondrial metabolic changes in senescent human fibroblasts

  • Ghneim, Hazem K.;Alfhili, Mohammad A.;Alharbi, Sami O.;Alhusayni, Shady M.;Abudawood, Manal;Aljaser, Feda S.;Al-Sheikh, Yazeed A.
    • The Korean Journal of Physiology and Pharmacology
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    • 제26권4호
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    • pp.263-275
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    • 2022
  • There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H2O2), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H2O2, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H2O2, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD+/NADH and NADP+/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.