• 미생물학회지
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• 제47권4호
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• pp.281-288
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• 2011

스트레스 반응에 관여하는 Saccharomyces cerevisiae 전사인자인 Msn2/4의 $C_2H_2$ zinc finger 부위와 아미노산 서열 유사성을 보이는 Aspergillus nidulans MsnA의 하위 유전자 획득을 위하여 msnA 결손 돌연변이체 또는 과발현 균주에서 야생주와 비교하여 차별적으로 발현되는 유전자(Differentially Expressed Gene, DEG)들을 분리하였다. 선별된 DEG들은 염기서열 결정을 통해 해당 유전자들을 동정하였고 이들 중 DEG6는 단당류 수송자(monosaccharide transporter)로 예측된 mstB 유전자로 밝혀졌다. mstB의 발현은 MsnA 과발현에 의하여 증가되었으며 MsnA는 in vitro에서 mstB 프로모터 부위에 직접적으로 결합하였다. MstB는 12개의 막결합 부위를 가지며 A. niger의 고친화성 단당류 수송자(high-affinity monosaccharide transporter)인 MstA와 80%의 높은 아미노산 서열 동일성을 보였다. mstB 결손 돌연변이체의 표현형은 야생주와 유사하였으나 MstB가 과발현된 균주는 낮은 당 농도인 0.1% glucose 배지에서 유성생식 기관인 cleistothecia의 형성이 증가하였다. 이러한 결과는 단당류 수송자인 MstB가 유성분화 과정에서 요구되는 당의 수송에 관여하고 있음을 시사한다.

• Journal of Applied Biological Chemistry
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• 제62권4호
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• pp.417-424
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• 2019

이 연구의 목적은 당 대사에 대한 삼색싸리(Lespedeza maximowiczii var. tricolor Nakai; LMTN)의 효과를 조사하는 것이다. LMTN 추출물은 대조군과 비교하여 3T3-L1 지방 세포에서 당 섭취능 및 지질축적을 유의하게 향상시켰다. 또한, 3T3-L1 지방 세포에서 LMTN 추출물은 퍼옥시좀 증식제 활성화 수용체(PPAR)γ, 인슐린수용체기질-1 (IRS-1) 및 포도당수송체(GLUT)4의 단백질 발현을 유의하게 증가시켰다. LMTM 추출물의 당 섭취능 또는 인슐린 신호 전달계의 조절 효과는 양성 대조물질인 트로글리타존 또는 피니톨보다 낮았지만 PPARγ 단백 활성화는 증가하였다. 또한, LMTM 추출물은 인슐린 유사효과를 나타냈다. db/db 마우스에서, LMTN 추출물(250 mg/kg BW)은 물과 식이 섭취량, 혈당, 중성지방과 총 콜레스테롤 함량을 유의적으로 감소시켰다. 더불어 지방과 근육조직에서의 PPARγ 및 GLUT4 mRNA의 발현도 LMTN 추출물 투여군에서 유의적으로 증가되었다. 따라서, 본 연구의 결과는 LMTN 추출물이 3T3-L1 지방세포 및 db/db 마우스에서 PPARγ 및 인슐린 유사효과를 통해 당 대사를 조절하는 것으로 밝혀졌다.

• The Korean Journal of Physiology and Pharmacology
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• 제20권2호
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• pp.153-160
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• 2016

The objective was to investigate the hypoglycemic action of catalpol in spontaneous diabetes db/db mice. 40 db/db mice were randomly divided into five groups: model control gourp; db/db plus catalpol 40, 80, 120 mg/kg body wt. groups and db/db plus metformin 250 mg/kg group. Age-matched db/m mice were selected as normal control group. The mice were administered with corresponding drugs or solvent by gavage for 4 weeks. The oral glucose tolerance test was carried out at the end of $3^{rd}$ week. After 4 weeks of treatment, the concentrations of fasting blood glucose (FBG), glycated serum protein (GSP), insulin (INS), triglyceride (TG), total cholesterol (TC) and adiponection (APN) in serum were detected. The protein expressions of phosphorylation-$AMPK{\alpha}$1/2 in liver, phosphorylation-$AMPK{\alpha}$1/2 and glucose transporter-4 (GLUT-4) in skeletal muscle and adipose tissues were detected by western blot. Real time RT-PCR was used to detect the mRNA expressions of acetyl-CoA carboxylase (ACC) and Hydroxymethyl glutaric acid acyl CoA reductase (HMGCR) in liver. Our results showed that catalpol could significantly improve the insulin resistance, decrease the serum concentrations of INS, GSP, TG, and TC. The concentrations of APN in serum, the protein expression of phosphorylation-$AMPK{\alpha}$1/2 in liver, phosphorylation-$AMPK{\alpha}$1/2 and GLUT-4 in peripheral tissue were increased. Catalpol could also down regulate the mRNA expressions of ACC and HMGCR in liver. In conclusion, catalpol ameliorates diabetes in db/db mice. It has benefit effects against lipid/glucose metabolism disorder and insulin resistance. The mechanism may be related to up-regulating the expression of phosphorylation-$AMPK{\alpha}$1/2.

• Archives of Pharmacal Research
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• 제25권6호
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• pp.923-931
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• 2002

Sopungsungi-won (SP) is a known for\mula for senile constipation and diabetes mellitus, based on traditional Korean medicine. The preventive effect of SP on the development of overt diabetes in Zucker diabetic fatty (ZDF) rats was evaluated. When administered orally through a diet for 8 weeks, diabetic conditions such as hyperglycemia, polydipsia and hypertriglyceridemia were all ameliorated in SP-treated rats. In parallel with the onset and progression of hyperglycemia in the ZDF control rats; there was a marked decline in plasma insulin concentrations from 26.1 $\mu$U/ml, at age 7 weeks, to 14.8 $\mu$U/ml at age 15 weeks. In the SP-treated rats, however, the plasma insulin concentrations did not decline, and SP at a dose of 5 g/kg significantly increased the insulin levels to 31.9 $\mu$U/ml. Early normalization of plasma insulin and a retained ability to subsequently increase plasma insulin were indicative of a pancreatic $\beta$ cell protective action by the SP for\mula. In addition, expressions of an insulin-responsive gene and corresponding protein, glucose transporter 4 (GLUT4), in skeletal \muscle, were also determined in SP- and rosiglitazone-treated ZDF rats. mRNA and protein levels of GLUT4 in SP-treated rats were upregulated in a dose dependent manner. Furthermore, when ZDF rats were treated with 2 g/kg of the SP for\mula, the activity of glucose-6-phosphatase was decreased by 49%, whereas the activity of glucokinase was increased by 196%, compared to the ZDF control rats. Taken together, these data provide evidence that the SP for\mula markedly lowered the plasma glucose levels, probably through an effect not only on improvement of insulin action, but through a combined sti\mulation of glycolysis and an inhibition of gluconeogenesis in the liver, and also suggest the validity of SP's clinical use in the treatment of type 2 diabetes mellitus following further toxicological investigation.

• 생명과학회지
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• 제19권12호
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• pp.1802-1807
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• 2009

Platycodi radix의 주요 성분으로 항염증, 항고지혈 및 항종양 등 다양한 약리적 기능을 가지는 platycodin D는 최근 비만 및 지방세포형성(adipogenesis)을 억제하는 효과를 가진다고 보고되고 있다. 본 연구에서는 adipogenesis의 상위단계에 위치한 다양한 pro-adipogenic regulators와 anti-adipogenic regulators의 발현이 platycodin D에 의해 어떻게 변화되는지 분석하였다. Real-time PCR을 이용한 mRNA 발현의 정량적 분석에서 adipogenesis의 marker라 불리는 ADIPOQ와 GLUT4의 mRNA 발현은 platycodin D의 처리에 의해 유의적으로 감소되었다. 또한 terminal marker의 발현을 조절하는 PPAR$\gamma$와 C/EPB$\alpha$의 mRNA 발현 역시 platycodin D에 의해 유의하게 억제되었다. Platycodin D의 지방세포 억제 효과에 대한 상세한 분자적 메커니즘을 규명하기 위해, PPAR$\gamma$와 C/EPB$\alpha$의 상위 조절자들의 mRNA 발현 변화를 분석하였다. Pro-adipogenic regulators에 대한 platycodin D의 효과를 분석한 결과, C/EBP$\beta$와 C/EPB$\delta$의 mRNA 발현은 platycodin D에 의해 변화가 없었던 반면, KROX20과 KLF15의 mRNA 발현은 각각 초기 분화(2일)와 후기 분화(4일)에서 platycodin D에 의해 유의한 감소를 보였다. 또한, 대표적인 anti-adipogenic regulators인 CHOP의 mRNA 발현은 초기분화에서 platycodin D에 의해 유의하게 증가한 반면, 또 다른 anti-adipogenic regulators인 C/EBP$\gamma$의 mRNA 발현은 platycodin D에 의해 영향을 받지 않았다. 따라서 adipogenesis 과정에서 platycodin D는 pro-adipogenic regulators인 KROX20, KLF15와 anti-adipogenic regulator인 CHOP의 mRNA 발현에 영향을 주어 PPAR$\gamma$와 C/EPB$\alpha$를 조절하는 것으로 보여진다. 결론적으로, platycodin D에 의한 adipogenesis 억제 효과는 KROX20, KLF15 등의pro-adipogenic regulator와 CHOP 등의 anti-adipogenic regulator의 상호작용을 통해 나타나는 결과라 사료된다.

• Journal of Ginseng Research
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• 제41권3호
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• pp.257-267
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• 2017

Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

• Genomics & Informatics
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• 제12권4호
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• pp.240-246
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• 2014

Mutation in HNF1B, the hepatocyte nuclear factor-$1{\beta}$ (HNF-$1{\beta}$) gene, results in maturity-onset diabetes of the young (MODY) 5, which is characterized by gradual impairment of insulin secretion. However, the functional role of HNF-$1{\beta}$ in insulin secretion and glucose metabolism is not fully understood. We identified a family with early-onset diabetes that fulfilled the criteria of MODY. Sanger sequencing revealed that a heterozygous P159L (CCT to CTT in codon 159 in the DNA-binding domain) mutation in HNF1B was segregated according to the affected status. To investigate the functional consequences of this HNF1B mutation, we generated a P159L HNF1B construct. The wild-type and mutant HNF1B constructs were transfected into COS-7 cells in the presence of the promoter sequence of human glucose transporter type 2 (GLUT2). The luciferase reporter assay revealed that P159L HNF1B had decreased transcriptional activity compared to wild-type (p < 0.05). Electrophoretic mobility shift assay showed reduced DNA binding activity of P159L HNF1B. In the MIN6 pancreatic ${\beta}$-cell line, overexpression of the P159L mutant was significantly associated with decreased mRNA levels of GLUT2 compared to wild-type (p < 0.05). However, INS expression was not different between the wild-type and mutant HNF1B constructs. These findings suggests that the impaired insulin secretion in this family with the P159L HNF1B mutation may be related to altered GLUT2 expression in ${\beta}$-cells rather than decreased insulin gene expression. In conclusion, we have identified a Korean family with an HNF1B mutation and characterized its effect on the pathogenesis of diabetes.

• Journal of Dairy Science and Biotechnology
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• 제39권2호
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• pp.78-85
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• 2021

본 연구는 고지방식이 마우스에서 체중 감소, 혈당 감소 및 인슐린 저항성 개선 효과를 가지는 LNS1 균주의 간 내 글리코겐 함량에 미치는 영향을 조사하여 고지방식이에 의한 비정상적인 글리코겐 대사 개선을 위한 활용 가능성을 검토하고자 실시하였다. LNS1을 12주간 경구 투여한 고지방식이 마우스의 간에서 포도당 운반체 단백질인 GLUT2와 글리코겐 합성의 주요 효소인 GCK, GYS2의 유전자 발현 변화를 확인한 결과, LNS1의 경구 투여는 고지방식이 마우스에 비해 GLUT2와 GYS2의 유전자 발현을 각각 약 2배, 1.8배 증가시켰으며, GCK의 발현에는 영향을 주지 않는 것으로 확인되었다. 또한, GCK의 regulatory unit으로 작용하여 GCK의 활성을 억제하는 GCKR와 글리코겐 분해 과정의 주요 효소인 G6PC의 발현은 LNS1 투여에 의해 HFD마우스에 비해 각각 약 53%, 32% 감소함을 보였다. 간 조직에서의 결과와 마찬가지로 HepG2 세포에 LNS1-CM의 처리는 GLUT2와 GYS2의 유전자 발현을 약 1.9배, 2배 증가시켰으며, GCK의 발현 변화에는 영향을 주지 않는 것으로 확인되었다. GCKR과 G6PC의 유전자 발현 또한 LNS1-CM 처리에 의해 각각 77%, 47% 감소함을 보였다. 또한, 간 조직 내 글리코겐 함량은 고지방식이와 LNS1 투여를 병행한 마우스에서 고지방식이 마우스에 비해 약 1.5배 증가한 것으로 조사되었다. 위의 결과들을 종합해 볼 때, LNS1은 GLUT2, GYS2, GCKR와 G6PC의 발현 조절을 통해 간 조직내 글리코겐 함량을 증가시켜 고지방식이에 의한 글리코겐 대사 이상을 개선시키는 효과를 가지는 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권4호
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• pp.651-661
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• 2020

Objective: We hypothesized that Cr source can alter adipogenic-related transcriptional regulations and cell signaling. Therefore, the objective of the study was to evaluate the biological effects of chromium acetate (CrAc) on bovine intramuscular (IM) and subcutaneous (SC) adipose cells. Methods: Bovine preadipocytes isolated from two different adipose tissue depots; IM and SC were used to evaluate the effect of CrAc treatment during differentiation on adipogenic gene expression. Adipocytes were incubated with various doses of CrAc: 0 (differentiation media only, control), 0.1, 1, and 10 μM. Cells were harvested and then analyzed by real-time quantitative polymerase chain reaction in order to measure the quantity of adenosine monophosphate-activated protein kinase-α (AMPK-α), CCAAT enhancer binding protein-β (C/EBPβ), G protein-coupled receptor 41 (GPR41), GPR43, peroxisome proliferator-activated receptor-γ (PPARγ), and stearoyl CoA desaturase (SCD) mRNA relative to ribosomal protein subunit 9 (RPS9). The ratio of phosphorylated-AMPK (pAMPK) to AMPK was determined using a western blot technique in order to determine changing concentration. Results: The high dose (10 μM) of CrAc increased C/EBPβ, in both IM (p = 0.02) and SC (p = 0.02). Expression of PPARγ was upregulated by 10 μM of CrAc in IM but not in SC. Expression of SCD was also increased in both IM and SC with 10 μM of CrAc treatment. Addition of CrAc did not alter gene expression of glucose transporter 4, GPR41, or GPR43 in both IM and SC adipocytes. Addition of CrAc, resulted in a decreased pAMPKα to AMPKα ration (p<0.01) in IM. Conclusion: These data may indicate that Cr source may influence lipid filling in IM adipocytes via inhibitory action of AMPK phosphorylation and upregulating expression of adipogenic genes.

• 동의생리병리학회지
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• 제17권2호
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.