• 한국컴퓨터정보학회논문지
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• 제29권1호
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• pp.187-194
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• 2024

본 연구에서는 산딸기 잎의 항비만 기능성 소재로서의 가능성을 확인하기 위하여 3T3-L1 지방전구세포에 산딸기 잎 추출물을 처리한 후 중성지방 함량을 측정하고 지방생성과 관련된 여러 단백질의 발현 양상을 측정하였다. 산딸기 잎은 열수로 추출·농축하여 시료로 사용하였으며 추출물의 수율은 4.76%였다. 산딸기 잎 추출물에 의한 3T3-L1 세포 생존율을 측정한 결과, 1,000 ㎍/mL 농도에서 13%의 성장 저해를 확인할 수 있었다. 이에 본 연구에서 시료 처리 농도를 500 ㎍/mL까지 처리하여 실험을 진행하였다. 세포 내 중성지방의 생성은 농도의존적으로 감소하는 경향을 나타내었으며, 감소율은 대조군에 비하여 농도 100, 300, 500 ㎍/mL에서 각각 28.7, 40.8, 51.6%를 나타내었다. 또한 PPARγ, C/EBPα 및 SREBP-1c, FAS 및 ACC의 발현 억제, p-AMPK 발현 증가로 지방합성이 억제되는 것을 확인하였다. 이상의 결과, 산딸기 잎 추출물이 지방세포 분화 및 지방생성 관련 인자들을 조절함으로서 비만 개선 소재로서의 활용이 가능할 것으로 판단된다.

• 한방비만학회지
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• 제24권1호
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• pp.1-12
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• 2024

Objectives: Ephedrae herba (EH) and Coicis semen (CS) has been frequently prescribed for the treatment of obesity. However, effects of combinational extracts of these two herbs on non-alcoholic fatty liver disease are unknown. The aim of the present study was to investigate the effects of EH and CS on lipid accumulation and glucose absorption in free fatty acids (FFAs) or palmitic acid (PA)-treated HepG2 cells. Methods: Five samples of EH and CS were extracted by combination ratios (S1=0:100, S2=25:75, S3=50:50, S4=75:25, S5=100:0). Oil Red O staining was used to measure lipid accumulation in FFAs-induced steatosis cells. Intracellular triglycerides and total cholesterol levels were measured in FFAs-induced steatotic HepG2 cells. In PA-treated cells, intracellular 2-NBDG was detected using a fluorescence microplate reader and flow cytometry. Phosphorylation of key metabolism-related factors of AMP-activated protein kinase and acetyl-CoA carboxylase, expression of key lipid synthesis-related factors carnitine palmitoyltransferase 1 alpha (CPT1α), sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT enhancer binding protein alpha (C/EBPα) were confirmed by western blot. Results: Treatment of EH-CS combination in the FFAs-induced steatotic HepG2 cells significantly reduced lipid accumulation. As the relative ratio of Ephedrae herba increased, the lipid-lowering effects of the combination were increased. However, S1 and S5 of Ephedrae herba and Coicis semen did not significantly reduce triglycerides and total cholesterol induced by FFAs. However, the combination of Ephedrae herba and Coicis semen restored glucose absorption in PA-induced HepG2 cells. Major makers of SREBP1, PPARγ, C/EBPα, and CPT1α expression tended to decrease with EH ratio. Conclusions: The EH-CS combination has advantages over sole EH and CS extracts in improving lipid and glucose metabolism in liver steatosis models.

• 한국자원식물학회지
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• 제37권4호
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• pp.307-313
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• 2024

본 연구는 추출용매에 따른 세잎쥐손이풀 추출물의 중성지방 조절에 대한 논문으로 3T3-L1 지방전구세포를 이용해 지방세포로 분화유도한 뒤 lipid accumulation, triglyceride contents, PPARγ, C/EBPα, FAS, SCD-1 및 AMPK 단백질 발현을 확인하였다. 실험결과 다섯가지 조건의 세잎쥐손이풀 추출물 중 NG-GT-T3L lipid accumulation과 triglyceride contents의 가장 높은 억제 효능을 확인하였다. NG-GT-T3L 추출물의 세포내 지방 축적 억제는 지방세포 내 adipogenesis와 lipogenesis를 조절하는 효소 조절을 통해 나타난 결과로 판단된다. 따라서 세잎쥐손이풀 추출물은 PPARγ, C/EBPα, FAS, SCD-1를 억제시키고, AMPK를 증가시켜 세포내 중성지방을 조절함으로서 체지방 감소 후보 소재로의 사용이 가능할 것으로 사료된다.

• 생명과학회지
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• 제25권10호
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• pp.1081-1090
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• 2015

본 연구는 우리나라에서 재배한 삼채 추출물의 주요생리활성 물질을 2형 당뇨 쥐에 투여하였을 때 인슐린 저항성 개선효과와 췌장에서 β-cell의 형태 및 간 조직의 2형 당뇨 관련 단백질의 발현을 확인 함으로써 그 메카니즘을 확인코자 하였다. 특히 TZDS당뇨약 계열로 현재 시판중인 actoz와 활성을 비교 함으로써 그 유효성 정도를 살펴보고자 본 실험을 디자인하였다. 먼저 식이섭취량, 물 섭취량의 경우, Control군은 2형 당뇨 특징인 다식, 다뇨, 다갈의 형태로 증가하였으나, 삼채 추출물 투여로 식이 및 물 섭취량이 감소하는 경향을 보였다. 식이 투여 2시간 후 혈당변화와 OGTT 검사결과, 2형 당뇨인 Control군에서 혈당의 농도가 가장 높았으나 삼채 추출물 8주 경구투여로 인해 안정화되는 것을 확인할 수 있었으며, serum의 glucose농도 또한 삼채 추출물 투여로 2형당뇨 control군에 비해 혈당이 감소한 것을 확인할 수 있었다. HbA1c (당화혈색소)는 2형 당뇨군(control)에서 9% 수치 범위로 고혈당 범위였으나, 삼채 추출물 섭취로 인하여 Positive control군(actoz 투여군) 7% 수준으로 감소하는 경향을 확인할 수 있었으며, insulin농도는 2형 당뇨인 Control군에서 insulin 농도가 1 ng/ml로 가장 낮은 농도였으며, 삼채 추출물 및 SAC 1.5 ng/ml수준으로 insulin수치가 상승하는 것을 확인할 수 있었다. 혈중 triglyceride, total cholesterol 및 LDL-cholesterol 수치 모두 시료 투여군에서 혈중 지질관련 인자의 농도가 감소하는 것을 확인할 수 있었으며, 특히 AH 400군의 경우 triglyceride, total cholesterol 및 LDL-cholesterol 농도가 각각 482.86±13.40 mg/dl, 163.56±13.24 mg/dl, 142.04±3.24 mg/dl로 대조군에 비해 약 50% 감소한 결과를 나타내었다. 간장 조직 중 지질 과산화의 함량의 경우, 2형 당뇨인 Control군에서 467.74±1.12 MDA nmole/g of tissue로 삼채 추출물 투여(AH 400)로 319.61±26.98 MDA nmole/g of tissue 로 Normal군과 비슷한 수준으로 혈중 lipid peroxide의 함량이 감소하는 것을 확인할 수 있었다. 간장조직을 H&E염색한 결과, Normal군에 비하여 2형 당뇨인 Control군의 간 조직에서 많은 지방적이 나타나 지방간이 확인되었으나, Positive control 및 삼채 추출물 투여에 의해 지방적의 크기와 수가 많이 줄어드는 결과를 얻어 조직학적 검사상 지방간의 소견이 현저히 호전되었으며, 췌장에서 insulin 면역화학 염색 결과 2형 당뇨로 췌도 세포가 거의 손상된 것과는 대조적으로, 삼채 추출물을 투여한 결과 정상군과 유사한 형태의 췌도 세포가 확인되었으며 췌도 세포의 형태가 다양한 정도의 과 형성을 보이는 것이 확인되었다. 또한 간장 조직의 PPARγ, PDK4, SREBP-1, FOX0 1의 발현을 확인한 결과, 발현이 2형 당뇨인 Control군에서 모두 발현이 증가하였으나, 삼채 추출물 투여로 인해 발현 정도가 낮게 나타나 앞서 확인하였던 glucose 및 HbA1c 및 insulin농도의 결과와 상통함을 확인하였다. 이들의 결과로 미루어 볼 때, 삼채 메탄올 추출물의 8주간 경구 투여로 인하여 2형 당뇨에 관여하는 PPARγ 단백질 및 비만 관련 단백질 인자를 조절하였으며, insulin농도 및 식후 혈당조절에도 도움을 주는 것을 확인할 수 있었다. 또한, 췌장의 insulin 면역화학염색 결과, 췌도 세포 형태 및 insulin 합성에도 긍정적인 역할을 하는 것으로 미루어 보아 2형 당뇨에 대한 치료효과가 큰 것으로 사료 된다.

• Biomolecules & Therapeutics
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• 제23권3호
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• pp.218-224
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• 2015

Endocannabinoids can affect multiple cellular targets, such as cannabinoid (CB) receptors, transient receptor potential cation channel, subfamily V, member 1 (TRPV1) and peroxisome proliferator-activated receptor ${\gamma}$($PPAR{\gamma}$). The stimuli to induce adipocyte differentiation in hBM-MSCs increase the gene transcription of the $CB_1$ receptor, TRPV1 and $PPAR{\gamma}$. In this study, the effects of three endocannabinoids, N-arachidonoyl ethanolamine (AEA), N-arachidonoyl dopamine (NADA) and 2-arachidonoyl glycerol (2-AG), on adipogenesis in hBM-MSCs were evaluated. The adipocyte differentiation was promoted by AEA whereas inhibited by NADA. No change was observed by the treatment of non-cytotoxic concentrations of 2-AG. The difference between AEA and NADA in the regulation of adipogenesis is associated with their effects on $PPAR{\gamma}$ transactivation. AEA can directly activate $PPAR{\gamma}$. The effect of AEA on $PPAR{\gamma}$ in hBM-MSCs may prevail over that on the $CB_1$ receptor mediated signal transduction, giving rise to the AEA-induced promotion of adipogenesis. In contrast, NADA had no effect on the $PPAR{\gamma}$ activity in the $PPAR{\gamma}$ transactivation assay. The inhibitory effect of NADA on adipogenesis in hBM-MSCs was reversed not by capsazepine, a TRPV1 antagonist, but by rimonabant, a $CB_1$ antagonist/inverse agonist. Rimonabant by itself promoted adipogenesis in hBM-MSCs, which may be interpreted as the result of the inverse agonism of the $CB_1$ receptor. This result suggests that the constantly active $CB_1$ receptor may contribute to suppress the adipocyte differentiation of hBM-MSCs. Therefore, the selective $CB_1$ agonists that are unable to affect cellular $PPAR{\gamma}$ activity inhibit adipogenesis in hBM-MSCs.

• BMB Reports
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• 제49권2호
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• pp.111-115
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• 2016

Caffeine has been proposed to have several beneficial effects on obesity and its related metabolic diseases; however, how caffeine affects adipocyte differentiation has not been elucidated. In this study, we demonstrated that caffeine suppressed 3T3-L1 adipocyte differentiation and inhibited the expression of CCAAT/enhancer binding protein (C/EBP)α and peroxisome proliferator-activated receptor (PPAR)γ, two main adipogenic transcription factors. Anti-adipogenic markers, such as preadipocyte secreted factor (Pref)-1 and Krüppel-like factor 2, remained to be expressed in the presence of caffeine. Furthermore, 3T3-L1 cells failed to undergo typical mitotic clonal expansion in the presence of caffeine. Investigation of hormonal signaling revealed that caffeine inhibited the activation of AKT and glycogen synthase kinase (GSK) 3 in a dose-dependent manner, but not extracellular signal-regulated kinase (ERK). Our data show that caffeine is an anti-adipogenic bioactive compound involved in the modulation of mitotic clonal expansion during adipocyte differentiation through the AKT/GSK3 pathway.

• Asian-Australasian Journal of Animal Sciences
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• 제27권1호
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• pp.10-18
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• 2014

The objective of this study was to investigate the correlation between cattle breeds and deposit of adipose tissues in different positions and the gene expressions of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), fatty acid synthase (FASN), and Acyl-CoA dehydrogenase (ACADM), which are associated with lipid metabolism and are valuable for understanding the physiology in fat depot and meat quality. Yanbian yellow cattle and Yan yellow cattle reared under the same conditions display different fat proportions in the carcass. To understand this difference, the expression of $PPAR{\gamma}$, FASN, and ACADM in different adipose tissues and longissimus dorsi muscle (LD) in these two breeds were analyzed using the Real-time quantitative polymerase chain reaction method (qRT-PCR). The result showed that $PPAR{\gamma}$ gene expression was significantly higher in adipose tissue than in LD in both breeds. $PPAR{\gamma}$ expression was also higher in abdominal fat, in perirenal fat than in the subcutaneous fat (p<0.05) in Yanbian yellow cattle, and was significantly higher in subcutaneous fat in Yan yellow cattle than that in Yanbian yellow cattle. On the other hand, FASN mRNA expression levels in subcutaneous fat and abdominal fat in Yan yellow cattle were significantly higher than that in Yanbian yellow cattle. Interestingly, ACADM gene shows greater fold changes in LD than in adipose tissues in Yan yellow cattle. Furthermore, the expressions of these three genes in lung, colon, kidney, liver and heart of Yanbian yellow cattle and Yan yellow cattle were also investigated. The results showed that the highest expression levels of $PPAR{\gamma}$ and FASN genes were detected in the lung in both breeds. The expression of ACADM gene in kidney and liver were higher than that in other organs in Yanbian yellow cattle, the comparison was not statistically significant in Yan yellow cattle.

• Asian-Australasian Journal of Animal Sciences
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• 제33권6호
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• pp.1012-1022
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• 2020

Objective: Caseins and fatty acids of milk are synthesized and secreted by the epithelial cells of the mammary gland. All-trans retinoic acid (ATRA), an active metabolite of vitamin A, has been shown to promote mammary development. This study was conducted to determine the effect of ATRA on casein synthesis and fatty acid composition in MAC-T cells. Methods: MAC-T cells were allowed to differentiate for 4 d, treated with ATRA (0, 1.0, 1.5, and 2.0 μM), and incubated for 3 d. We analyzed the fatty acid composition, the mRNA expression of casein and fatty acid synthesis-related genes, and the phosphorylation of casein synthesis-related proteins of MAC-T cells by gas chromatography, quantitative polymerase chain reaction, and western blotting, respectively. Results: In MAC-T cells, ATRA increased the mRNA levels of α_S1-casein and β-casein, janus kinase 2 (JAK2) and E74-like factor 5 of the signal transducer and activator of transcription 5 β (STAT5-β) pathway, ribosomal protein S6 kinase beta-1 (S6K1) and eukaryotic translation initiation factor 4E binding protein 1 of the mammalian target of rapamycin (mTOR) pathway, inhibited the mRNA expression of phosphoinositide 3-kinase and eukaryotic initiation factor 4E of the mTOR pathway, and promoted the phosphorylation of STAT5-β and S6K1 proteins. Additionally, ATRA increased the de novo synthesis of fatty acids, reduced the content of long-chain fatty acids, the ratio of monounsaturated fatty acids to saturated fatty acids (SFA), the ratio of polyunsaturated fatty acids (PUFA) to SFA, and the ratio of ω-6 to ω-3 PUFA. The mRNA levels of acetyl-CoA carboxylase 1, fatty acid synthase, lipoprotein lipase, stearoyl-CoA desaturase, peroxisome proliferator-activated receptor gamma, and sterol regulatory element-binding protein 1 (SREBP1) were enhanced by ATRA. Conclusion: ATRA promotes the synthesis of casein by regulating JAK2/STAT5 pathway and downstream mTOR signaling pathway, and it improves the fatty acid composition of MAC-T cells by regulating SREBP1-related genes.

• Asian-Australasian Journal of Animal Sciences
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• 제30권9호
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• pp.1215-1224
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• 2017

Objective: To assess the differences in ovarian transcriptomes in Shan Ma ducks between their peak and late stages of egg production, and to obtain new transcriptomic data of these egg-producing ducks. Methods: The Illumina HiSeq 2000 system was used for high throughput sequencing of ovarian transcriptomes from Shan Ma ducks at their peak or late stages of egg production. Results: Greater than 93% of the sequencing data had a base quality score (Q score) that was not less than 20 (Q20). From ducks at their peak stage of egg production, 42,782,676 reads were obtained, with 4,307,499,083 bp sequenced. From ducks at their late stage of egg production, 45,316,166 reads were obtained, with 4,562,063,363 bp sequenced. A comparison of the two datasets identified 2,002 differentially expressed genes, with 790 upregulated and 1,212 downregulated. Further analysis showed that 1,645 of the 2,002 differentially expressed genes were annotated in the non-redundant (NR) database, with 646 upregulated and 999 downregulated. Among the differentially expressed genes with annotations in the NR database, 696 genes were functionally annotated in the clusters of orthologous groups of proteins database, involving 25 functional categories. One thousand two hundred four of the differentially expressed genes with annotations in the NR database were functionally annotated in the gene ontology (GO) database, and could be divided into three domains and 56 categories. The three domains were cellular component, molecular function, and biological process. Among the genes identified in the GO database, 451 are involved in development and reproduction. Analysis of the differentially expressed genes with annotations in the NR database against the Kyoto encyclopedia of genes and genomes database revealed that 446 of the genes could be assigned to 175 metabolic pathways, of which the peroxisome proliferator-activated receptor signaling pathway, insulin signaling pathway, fructose and mannose metabolic pathways, gonadotropin releasing hormone signaling pathway and transforming growth factor beta signaling pathway were significantly enriched. Conclusion: The differences in ovarian transcriptomes in Shan Ma ducks between their peak and late stages of egg production were elucidated, which greatly enriched the ovarian transcriptomic information of egg-producing ducks.

• 한국식품과학회지
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• 제47권1호
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• pp.111-118
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• 2015

본 연구는 항비만 소재로 연구된 도라지와 울금 소재를 대상으로 발효 전과 후 3T3-L1 지방세포의 분화 억제효능을 확인하고자 하였다. 그 결과 발효 전 도라지와 울금은 3T3-L1 세포의 분화에 영향을 미치지 못하는 것을 확인할 수 있었다. Apsergillus oryzae 발효를 통해 만들어진 소재들은 도라지 발효물의 플라보노이드 증가를 제외하면 모든 소재에서 폴리페놀과 플라보노이드의 함량 및 항산화 활성이 감소하는 것을 확인할 수 있었다. 그러나 발효 전과 다르게 발효 후 도라지, 울금 및 도라지와 울금 혼합물의 발효물에서 3T3-L1 세포의 분화억제능이 oil red O 염색법을 통해 관찰되었으며, mRNA 발현 확인을 통해 SREBP-1c와 $PPAR{\gamma}$의 발현을 억제하여 분화를 억제할 수 있음을 확인하였다. 이는 Apsergillus oryzae 발효과정이 도라지와 울금의 항비만효과를 개선할 수 있으며, 발효과정을 통해 새로운 항비만 유효성분이 생성되었음을 예측할 수 있었다. 본 연구결과를 통해 Apsergillus oryzae 발효공정이 도라지와 울금의 기능을 향상시켜 우수한 기능성 제품의 개발에 유용하게 활용될 수 있음을 확인할 수 있었으며, 개발공정의 기초자료를 제공할 것으로 사료된다.