• Animal Bioscience
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• 제36권2호
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• pp.200-208
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• 2023

Objective: Muscle acetylcholine receptors have five alpha subunits (α, β, δ, ε, or γ), and cholinergic receptor nicotinic gamma subunit (CHRNG) is the γ subunit. It may also play an essential role in biological processes, including cell differentiation, growth, and survival, while the role of CHRNG has not been studied in the literature. Therefore, the purpose of this study is to clarify the effect of CHRNG on the proliferation and differentiation of bovine preadipocytes. Methods: We constructed a CHRNG overexpression adenovirus vector and successfully overexpressed it on bovine preadipocytes. The effects of CHRNG on bovine preadipocyte proliferation were detected by Edu assay, cell counting Kit-8 (CCK-8), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), Western blot and other techniques. We also performed oil red O, RT-qPCR, Western blot to explore its effect on the differentiation of preadipocytes. Results: The results of Edu proliferation experiments showed that the number of EDU-positive cells in the overexpression group was significantly less. CCK-8 experiments found that the optical density values of the cells in the overexpression group were lower than those of the control group, the mRNA levels of proliferating cell nuclear antigen (PCNA), cyclin A2 (CCNA2), cyclin B1 (CCNB1), cyclin D2 (CCND2) decreased significantly after CHRNG gene overexpression, the mRNA levels of cyclin dependent kinase inhibitor 1A (CDKN1A) increased significantly, and the protein levels of PCNA, CCNB1, CCND2 decreased significantly. Overexpression of CHRNG inhibited the differentiation of bovine preadipocytes. The results of oil red O and triglyceride determination showed that the size and speed of lipid droplets accumulation in the overexpression group were significantly lower. The mRNA and protein levels of peroxisome proliferator activated receptor gamma (PPAR class="checkNonKBPoint">γ), CCAAT enhancer binding protein alpha (CEBPα), fatty acid binding protein 4 (FABP4), fatty acid synthase (FASN) decreased significantly. Conclusion: Overexpression of CHRNG in bovine preadipocytes inhibits the proliferation and differentiation of bovine preadipocytes.

• 대한의생명과학회지
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• 제12권2호
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• pp.73-79
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• 2006

Nebulin is a giant ($600{\sim}900$ kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin filamints in vertebrate skeletal muscles. Recently, There is an evidence that the nebulin also expressed in non muscle tissue, brain and liver. We identified a new isoform of nebulin from adult brain library by PCR screening. It contains two simple-repeats exon 165, 166 and linker-repeats exon $154{\sim}161$ except exon 159. The nebulin modules M160 to M170 (exon 150 to exon 161) has been shown to bind desmin. In mature striated muscle, desmin intermediate filaments surround Z-discs and link individual myofibrils laterally at their Z-discs and to other intracellular structures, including the costameres and the intercalated discs of the sarcolemma, sarcoplasmic reticulum, mitochondria, T-tubules, and nuclei. Therefore, it is an interesting possibility that the differential splice pathways within the linker region of nebulin modify the affinity of nebulin's interaction with desmin. The specific interactions of nebulin and desmin were confirmed in vivo by yeast two hybrid experiments. To verify in the cellular level the interaction between nebulin isoform and desmin, we transfected COS-7 cell with EGFP-tagged nebulin and DsRed-tagged desmin. Based on evidence showing that despite exon 159 was deleted, the new isoform of nebulin was interact with desmin. This suggest that nebulin in brain may interact with another intermediate filament. The conservation of these ligand-binding capacity in brain and skeletal nebulins suggest that nebulins may have conserved roles in brain and skeletal muscle.

• 대한의생명과학회지
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• 제13권3호
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• pp.207-212
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• 2007

The purpose of this preliminary study is to improve the efficiency of gene transfer of nonviral plasmid DNA by in vivo electroporation in ischemic hindlimb muscle, tibialis anterior. Hindlimb ischemic model was aseptically made by excision of left femoral artery. Each $50\;{\mu}g$ of pEGFP-C1 and pGL3-control in $100\;{\mu}l$ 0.9% NaCl was injected in tibialis anterior muscle. In vivo electroporation was applied on the same site with 10 mm-distance 2 needle array electrodes and ECM830. In 3 groups of normal rat with different electric field strength 0, 200 and 800 V/cm, the expression of pEGFP-C1 was comparatively evaluated. In 8 groups of normal rats, the expression of pGL3-control was evaluated in 0, 40, 50, 80, 100, 150, 200 and 300 V/cm of electric field strength. In 5 groups of ischemic models, the expression of pGL3-control was analyzed on 0, 4, 7, 10 and 14 days elapsed after making ischemic models. In 9 groups of ischemic rats, the expression of pGL3-control was analyzed in the electric field strength 0, 60, 70, 80, 100, 150, 200, 250 and 300 V/cm. GFP expressions in normal tibialis anterior were high in the extent and degree in order of electric field strength of 200, 800 and 0 V/cm. Luciferase value was highest in $50{\sim}100\;V/cm$ electric field strength. In the case of ischemic models, luciferase expression was significantly increasing in the order elapsed time after making the model. The degree of luciferase expression was higher in cases of application of in vivo electroporation than in that of non-application and was highest in $100{\sim}150\;V/cm$. In conclusion, in vivo electroporation is effective in transfer and expression of plasmid DNA in normal and ischemic tibialis anterior of rat.

• Applied Biological Chemistry
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• 제30권3호
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• pp.210-218
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• 1987

전보에서 무화과 latex에서 ficin을 분리 정제한 바 있다. 본 연구에 있어서 조 ficin을 우육에 처리하여 그의 연육제로서의 단백질 분해효과를 살피기 위하여 ficin을 농도별로 우육에 작용했을 때 생기는 유리아미노산 및 질소화합물의 함량을 측정 하였으며 이들을 숙성 및 조리시의 함량과 비교검토하였으며 그 결과는 다음과 같다. 유리 아미노산의 함량은 $1^{\circ}C$ 및 $8^{\circ}C$에서 숙성시 처리온도 및 시간에 따라 증가하였고 생우육에 ficin 처리시는 더욱 증가하였으며 증가율 또한 현저하게 높았다. 조리후 ficin 처리의 경우는 "삶기", "찌기", "굽기"의 경우중 찌기가 유리아미노산이 가장 많이 증가하였으며 또한 아미노산 증가량은 숙성중($1^{\circ}C$, 3일) 생육에 ficin 처리 (0.1% 8시간) 조리후 ficin 처리(0.1% 2시간)을 비교하였을 때 각각 13%, 292%, 137%로서 ficn 처리시의 두 group이 숙성중인 것보다 월등히 증가하였다. 총질소, 비단백태질소 및 암모니아태질소의 함량 변화는 $1^{\circ}C$ 및 $8^{\circ}C$에서 숙성시 처리온도와 시간에 따라 증가하였으며 생우육에 ficin 처리시는 더욱 증가하였으며 그 증가율이 현저하게 높았다. 조리후 ficin의 처리 경우에는 "삶기", "찌기", "굽기"의 경우에서 "찌기"가 암모니아태질소를 제외하고는 그 수치가 가장 높았다. 또한 숙성중($1^{\circ}C$, 3일)인 생육과 ficin 처리시의 생육의 총질소에 대한 비단백태질소의 증가율을 비교하였을 때 ficin 처리한 생육이 숙성중인 생육보다 크게 증가하였다.

• 한국식품과학회지
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• 제49권2호
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• pp.192-202
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• 2017

황정 주정 추출물 ID1216의 고지방 식이 유도 비만 마우스에서의 체중 증가 억제 효과에 대한 분자생물학적 기전을 확인하고자 단백질과 mRNA 수준에서 지질 및 에너지 대사 관련 유전자들의 발현 변화를 관찰하였다. 본 연구에서 확인된 지표들 간의 상호 작용 및 ID1216의 조절 여부에 관해 Fig. 10에 나타내었다. 실험 결과 ID1216은 고지방 식이 유도 비만 마우스에서 체중 증가 억제를 나타내었으며, 비만 대사 관련 pathway의 상위 유전자로 사료되는 SIRT1과 AMPK의 발현을 조절하는 것으로 나타났다. 활성화된 SIRT1과 AMPK는 $PGC1{\alpha}$의 활성화에 관여하고, 이를 통해 열 발산 대사와 관련된 UCP 단백질과 핵 수용체 단백질인 $PPAR{\alpha}$의 발현이 백색지방, 갈색지방, 간 및 근육에서 증가되는 것이 확인되었다. 각 조직 별로 RT-PCR을 진행한 결과에서는 $PPAR{\alpha}$의 하위 유전자인 aP2, ACO, Acadl, Acadm, CPT1a, CPT1b의 mRNA 발현 수준을 향상시켜 주어 ID1216이 지방산 산화 대사인 ${\beta}$-oxidation의 활성화에 기여할 가능성을 보여주었다. 이와는 별개로 ID1216은 중성지질을 분해하는 것으로 알려진 ATGL의 mRNA 발현 또한 증가시키는 것으로 확인되었다. 본 연구를 통해 ID1216이 조직에 따라 지질 및 에너지 대사와 관련된 인자의 발현에 영향을 주는 체중 조절에 효과적인 소재임을 알 수 있었다. 또한 비만 치료제와의 기전적 차별성과 생약 특유의 섭취 안전성을 특징으로 하는 체중 또는 체지방 조절 기능성 소재로의 활용 가능성도 충분히 가지고 있음을 확인할 수 있었다.

• 생명과학회지
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• 제17권6호통권86호
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• pp.859-866
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• 2007

본 연구는 임신횐쥐에서 운동이 혈중 인슐린과 렙틴 농도, 그리고 골격근의 GLUT-4와 VAMP-2단백질 발현에 변화를 조사하기 위하여 수행되었다. 본 연구에서 비임신 횐쥐에 비해 임신 횐쥐에서 혈중 인슐린 농도가 유의하게 증가되었고, 렙틴 농도는 유의하게 감소하는 것으로 나타났다. 임신횐쥐에게 달리기 운동을 적용한 결과 임신으로 증가된 인슐린 농도를 유의하게 감소시키면서 렙틴 농도의 감소를 억제시키는 것으로 나타났다. 골격근에서 GLUT-4와 VAMP-2 단백질 발현량을 조사한 결과 비임신 횐쥐에 비해 임신 횐쥐에서 이러한 단백질 발현이 유의하게 감소되었지만, 임신 중 달리기운동을 경험한 횐쥐에서 GLUT-4와 VAMP-2 발현 모두 유의하게 증가되는 것으로 나타났다. 이와 잘이 임신으로 인한 혈중 인슐린 및 렙틴의 변화가 골격근에서 당을 근조직으로 흡수하는 신호 전달 경로와 수송체의 발현 손상과 연관되어있음을 보여주는 것이다. 하지만 임신 횐쥐에서 달리기 운동은 인슐린과 렙틴 농도의 변화를 완화시킴으로써 골격근의GLUT-4와 VAMP-2발현을 유의하게 증가시키는 것으로 나타났다.

• 대한의생명과학회지
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• 제18권3호
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• pp.227-236
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• 2012

Previous study showed that swimming improved obesity but was not through $PPAR{\alpha}$ activation in liver and skeletal muscle in high fat diet-fed female mice with functioning ovaries as an animal model of obese premenopausal women. Thus, this study was aimed at investigation of the effects of swimming on the promotion of health and its molecular mechanism in adipose tissue of high fat diet-fed female mice. Eight-week-old female C57BL/6J mice were randomly divided into two groups (a non-swim control group and a swim group, n=8/group). Mice in the swim group swam for 2 h daily for 6 weeks in water bath with temperature of $35{\pm}1^{\circ}C$. All the animals received high fat diet (45% kcal fat) for 6 weeks. Reverse transcription-polymerase chain reaction was used to elucidate the molecular mechanism. Female mice subjected to swimming had significantly decreased body weight gain and white adipose tissue mass compared with the female control mice. Histological studies illustrated that swimming decreases the hepatic lipid accumulation. As expected, swimming did not affect the expression of mRNA levels of peroxisome proliferator-activated receptor (PPAR) ${\alpha}$ and $PPAR{\alpha}$ target genes responsible for mitochondrial fatty acid ${\beta}$-oxidation, such as carnitine palmitoyltransgerase-1 and medium chain acyl-CoA dehydrogenase in the white adipose tissue. However, mice that underwent 6-weeks of swimming exercise had decreased the mRNA expression of lipogenic genes, such as sterol regulatory element-binding proteins-1C and fatty acid synthase in comparison to sedentary control mice, with decreased $PPAR{\gamma}$ target genes involved in adipocyte-specific marker genes, such as adipocyte fatty acid binding protein and leptin in the white adipose tissue. These results suggest that swimming can effectively prevent obesity induced by high fat diet-fed, in part through down-regulation of adipogenesis and lipogenesis in white adipose tissue of female obese mice. Moreover, these results suggest that swimming maybe contributing the promotion of health through regulation of adipogenesis and lipogenesis in overweight premenopausal women.

• 한국수산과학회지
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• 제42권1호
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• pp.26-33
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• 2009

This experiment was conducted to examine the utilization of added dietary selenium (Se) as an immune stimulant in juvenile olive flounder, Paralichthys olivaceus. Fish averaging $4.0{\pm}0.1\;g$ ($mean{\pm}SD$) were fed one of seven semi-purified diets containing 0.56, 1.07, 2.86, 4.56, 43.15, 90.71, or 161.74 mg of Se/kg ($Se_{0.56}$, $Se_{1.07}$, $Se_{2.86}$, $Se_{4.56}$, $Se_{43.2}$, $Se_{90.7}$ and $Se_{161.7}$, respectively) for 12 weeks, respectively. At the end of the feeding trial, the fish fed diets containing more than 43.2 mg of Se/kg showed above 90% mortality. There were no significant differences in weight gain, feed efficiency, specific growth rate, protein efficiency ratio, or hematological characteristics among the fish fed the $Se_{0.56}$, $Se_{l.07}$, $Se_{2.86}$, and $Se_{4.56}$ diets. Se concentrations of the gill, kidney, muscle and liver tissues occurred in dose-dependent manners. Alternative complement pathway activation and the chemiluminescene responses of the fish fed the $Se_{1.07}$ diet were significantly higher than those of the fish fed the other diets (P<0.05). These results indicate that the optimum dietary supplementation level of Selenium as selenoyeast could be 1.07 mg of Se/kg based on the non-specific immune responses of juvenile oilve flounder, Paralichthys olivaceus.

• 한국수산과학회지
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• 제42권5호
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• pp.411-416
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• 2009

Proximate compositions and fatty acid profiles of cultured and wild mackerel (Scomber japonicus) muscles were compared. Protein content ranged from approximately 16% to 18% and was higher in wild fish than in cultured ones. Lipid content was between two to four times higher in large and small cultured fish (20.1-20.5%) compared with same sized wild fish. The prominent non-polar lipid (NL) class in fish muscles was triglyceride, and additionally, free sterol was among the prominent NL classes in wild fish muscles. Prominent phospholipid (PL) classes in cultured and wild fish muscles were phosphatidylethanolamine and phosphatidylcholine, with the former being higher in cultured fish and the latter higher in wild fish. Prominent fatty acids of total lipid were 16:0, 18:1n-9, 22:6n-3 (docosahexaenoic acid, DHA), 20:5n-3 (eicosapentaenoic acid, EPA), 16:1n-7, 18:0 and 14:0, while 18:2n-6 was among the prominent fatty acids in cultured fish. The n-3 polyunsaturated fatty acids (PUFA, DHA+EPA) content (in mg/100 g of muscle tissue) was higher in cultured fish (2,711 mg in large fish and 2,572 mg in small fish) than in wild fish (2,431 mg in large fish and 1,398 mg in small fish). In conclusion, we have been able to demonstrate that cultured mackerel could also be a good sources of n-3 PUFA, such as DHA and EPA.

• Journal of Nutrition and Health
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• 제29권7호
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• pp.772-777
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• 1996

The purpose of this study was to investigate the effect of vitamin B2 deficiency on the utilization and recuperation of fuel in exercising rats. Thirthy six rats were fed either a vitamin B2 deficient diet (-B2) or a control diet (+B2) for 3 weeks and then subdivided into 3 groups : non-exercise group(NE), exercise group (ES), exercise and recuperation group (ER). ES group were exercised on treadmill (10$^{\circ}$, 0.5-0.8km/h) for 2 hours and ER group were recuperated three days with the respective diet after exercise. Glucose (GLU), glycogen(GLY), protein(PRO), triglyceride(TG) and free fatty acid(FFA) were compared in plasma(P), liver(L) and skeletal muscle(M) of rats. Compared to + B2 rats, in NE group, the level of P-GLU and L-GLU of -B2 rats was lower, L-PRO and M-GLY was higher and there were no differences in P-PRO, P-FFA, L-TG and M-TG. In ES group, the level of P-GLU was lower, P-FFA was higher and there were no differences in P-PRO, P-TG, L-GLU, L-TG, M-GLY, M-TG and M-PRO. In ER groups, the level of P-GLU and L-TG was lower, P-FFA was higher and there were no differences in P-PRO, P-TG, L-GLY, L-PRO, M-GLY, M-TG and M-PRO. These results suggest that a vitamin B2 deficiency may impair the utilization of stored fuel during exercise suggest that a vitamin B2 deficiency may impair the utilization of stored fuel during exercise and may lead a sluggish recuperation related to fuel stores after exercise.