• Journal of Ginseng Research
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• 제41권1호
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• pp.60-68
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• 2017

Background: Various Panax ginseng cultivars exhibit a range of diversity for morphological and physiological traits. However, there are few studies on diversity of metabolic profiles and genetic background to understand the complex metabolic pathway in ginseng. Methods: To understand the complex metabolic pathway and related genes in ginseng, we tried to conduct integrated analysis of primary metabolite profiles and related gene expression using five ginseng cultivars showing different morphology. We investigated primary metabolite profiles via gas chromatography-mass spectrometry (GC-MS) and analyzed transcriptomes by Illumina sequencing using adventitious roots grown under the same conditions to elucidate the differences in metabolism underlying such genetic diversity. Results: GC-MS analysis revealed that primary metabolite profiling allowed us to classify the five cultivars into three independent groups and the grouping was also explained by eight major primary metabolites as biomarkers. We selected three cultivars (Chunpoong, Cheongsun, and Sunhyang) to represent each group and analyzed their transcriptomes. We inspected 100 unigenes involved in seven primary metabolite biosynthesis pathways and found that 21 unigenes encoding 15 enzymes were differentially expressed among the three cultivars. Integrated analysis of transcriptomes and metabolomes revealed that the ginseng cultivars differ in primary metabolites as well as in the putative genes involved in the complex process of primary metabolic pathways. Conclusion: Our data derived from this integrated analysis provide insights into the underlying complexity of genes and metabolites that co-regulate flux through these pathways in ginseng.

• 한국자기공명학회논문지
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• 제12권1호
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• pp.14-25
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• 2008

To investigate the 3-bond connectivity of human brain metabolites by scalar coupling interaction through 2D-correlation spectroscopy (COSY) techniques using high field NMR spectroscopy. All NMR experiments were performed at 298K on Unity Inova 500 or 600 (Varian Inc.) equipped with a triple resonance probe head with z-shield gradient. Human brain metabolites were prepared with 10% $D_2O$. Two dimensional 2D COSY spectra were acquired with 4096 complex data points in $t_2$ and 128 or 256 increments in $t_1$ dimension. The spectral width was 9615.4 Hz and solvent suppression was achieved using presaturation using low power irradiation of the water resonance during 2s of relaxation delay. NMR data were processed using VNMRJ (Varian Instrument) software and all the chemical shifts were referenced to the methyl resonance of N-acetyl aspartate (NAA) peak at 2.0 ppm. Total 10 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), lactate (Lac), taurine (Tau), ${\gamma}$-aminobutyricacid (GABA), alanine (Ala) were included for major target metabolites. Symmetrical 2D-COSY spectra were successfully acquired. Total 14 COSY cross peaks were observed even though there were parallel/orthogonal noisy peaks induced by water suppression. Except for Cr, all of human brain metabolites produced COSY cross peaks. The spectra of NAA methyl proton at 2.02 ppm and Glu methylene proton ($CH_2(3)$) at 2.11 ppm and Gln methylene proton ($CH_2(3)$) at 2.14 ppm were overlapped in the similar resonance frequency between 2.00 ppm and 2.15 ppm. The present study demonstrated that in vitro 2D-COSY represented the 3-bond connectivity of human brain metabolites by scalar coupling interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2D-COSY study. Also it would be helpful to determine the molecular stereochemistry in vivo by using two-dimensional MR spectroscopy.

• Journal of Animal Science and Technology
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• 제63권3호
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• pp.531-544
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• 2021

Heat stress (HS) damages health and decreases performance variables in pigs, and if severe enough, causes mortality. However, metabolic changes under HS and recovery following HS are poorly understood. Therefore, this study was aimed to expose the essential mechanisms by which growing pigs respond to HS and the temporal pattern of plasma concentrations (PC) of amino acids (AAs) and metabolites. Crossbred male growing pigs were penned separately and allowed to adapt to thermal-neutral (TN) conditions (20℃ and 80% relative humidity; TN[-1D]). On the first day, all pigs were exposed to HS for 24 h (36℃ and 60% relative humidity), then to TN conditions for 5 days (TN[2D] to TN[5D]). All pigs had ad libitum access to water and 3 kg feed twice daily. Rectal temperature (RT) and feed intake (FI) were determined daily. HS pigs had higher RT (40.72℃) and lower (50%) FI than TN(-1D) pigs (p < 0.01). The PC of indispensable (threonine, valine, and methionine) and dispensable (cysteine and tyrosine) AAs were higher (p < 0.05) in HS than TN(-1D) pigs and remained increased during recovery time. Nonprotein α-aminobutyric acid and β-alanine concentrations were higher (p < 0.05) in HS than TN(-1D) pigs. The metabolite concentration of creatinine was higher (p < 0.01) under HS treatment than other treatments, but that of alanine and leucine remained increased (p < 0.05) through 5 d of recovery. In summary, some major differences were found in plasma AA profiles and metabolites between HS- and TN-condition pigs. This indicates that the HS pigs were forced to alter their metabolism, and these results provide information about mechanisms of acute HS responses relative to the recovery time.

• 생물환경조절학회지
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• 제31권4호
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• pp.332-342
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• 2022

케일(Brassica oleracea var. acephala)은 필수 아미노산, 비타민, 미네랄과 같은 수많은 영양소를 함유하고 특히 글루코시놀레이트가 풍부하기 때문에 전 세계적으로 가장 많이 소비되는 잎 채소 중 하나이다. 그러나 케일 품종 간의 글루코시놀레이트 합성과 관련된 유전자 발현에 대한 연구는 미비한 실정이다. 본 연구에서는 전사체 및 대사체 분석을 사용하여 식물공장에서 재배된 녹색(만추 및 맛짱) 및 적색 케일 품종(적곱슬)을 포함한 3 가지 케일 품종에서 글루코시놀레이트를 조사하였다. 재배 후 6주된 녹색 케일 품종의 생육 및 발달이 적색 케일 품종에 비해 높았다. High-performance liquid chromatography (HPLC) 분석에서 7가지 글루코시놀레이트를 분석하였다; 만추 품종에서는 5종의 글루코시놀레이트가, 맛짱과 적곱슬 품종에서는 4종의 글루코시놀레이트가 분류되었다. Glucobrassicin은 3가지 케일 품종에서 가장 높은 글루코시놀레이트 였다. 총 글루코시놀레이트 함량은 적곱슬 품종에서 가장 높았다. 전사체 분석에서는 8개의 유전자가 글루코시놀레이트 합성에 관여됨을 확인할 수 있었다. 이러한 결과는 케일 품종에 따라 글루코시놀레이트 함량과 축적 패턴이 다르다는 것을 시사한다.

• Journal of Ginseng Research
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• 제46권2호
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• pp.235-247
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• 2022

Background: Ginsenoside Rg3 is one of the main active ingredients in ginseng. Here, we aimed to confirm its protective effect on the heart function in transverse aortic coarctation (TAC)-induced heart failure mice and explore the potential molecular mechanisms involved. Methods: The effects of ginsenoside Rg3 on heart and mitochondrial function were investigated by treating TAC-induced heart failure in mice. The mechanism of ginsenoside Rg3 for improving heart and mitochondrial function in mice with heart failure was predicted through integrative analysis of the proteome and plasma metabolome. Glucose uptake and myocardial insulin sensitivity were evaluated using micro-positron emission tomography. The effect of ginsenoside Rg3 on myocardial insulin sensitivity was clarified by combining in vivo animal experiments and in vitro cell experiments. Results: Treatment of TAC-induced mouse models with ginsenoside Rg3 significantly improved heart function and protected mitochondrial structure and function. Fusion of metabolomics, proteomics, and targeted metabolomics data showed that Rg3 regulated the glycolysis process, and Rg3 not only regulated glucose uptake but also improve myocardial insulin resistance. The molecular mechanism of ginsenoside Rg3 regulation of glucose metabolism was determined by exploring the interaction pathways of AMPK, insulin resistance, and glucose metabolism. The effect of ginsenoside Rg3 on the promotion of glucose uptake in IR-H9c2 cells by AMPK activation was dependent on the insulin signaling pathway. Conclusions: Ginsenoside Rg3 modulates glucose metabolism and significantly ameliorates insulin resistance through activation of the AMPK pathway.

• Journal of Ginseng Research
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• 제47권1호
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• pp.44-53
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• 2023

Background: The genus Panax in the Araliaceae family has been used as traditional medicinal plants worldwide and is known to biosynthesize ginsenosides and phytosterols. However, genetic variation between Panax species has influenced their biosynthetic pathways is not fully understood. Methods: Simultaneous analysis of transcriptomes and metabolomes obtained from adventitious roots of two tetraploid species (Panax ginseng and P. quinquefolius) and two diploid species (P. notoginseng and P. vietnamensis) revealed the diversity of their metabolites and related gene expression profiles. Results: The transcriptome analysis showed that 2,3-OXIDOSQUALENE CYCLASEs (OSCs) involved in phytosterol biosynthesis are upregulated in the diploid species, while the expression of OSCs contributing to ginsenoside biosynthesis is higher in the tetraploid species. In agreement with these results, the contents of dammarenediol-type ginsenosides were higher in the tetraploid species relative to the diploid species. Conclusion: These results suggest that a whole-genome duplication event has influenced the triterpene biosynthesis pathway in tetraploid Panax species during their evolution or ecological adaptation. This study provides a basis for further efforts to explore the genetic variation of the Panax genus.

• Investigative Magnetic Resonance Imaging
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• 제12권2호
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• pp.100-106
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• 2008

본 연구는 in vivo 보다 더욱 정확하게 뇌 대사물질을 정량 분석하고자 고자장 NMR 장비 (500MHz; 11.74T)를 이용하여 동물의 뇌를 in vitro 환경에서 조사 및 분석하였다. 일반적으로 in vivo 실험은 생체 내부의 혈류나 조직의 비균질성으로 인한 자기공명분광법의 shimming에 악영향을 미치기 때문에 부정확한 결과를 산출할 수 있다. 그러나, in vitro 실험은 이에 비하여 균질한 샘플을 사용하고 보다 고자장에서 실험환경을 조성할 수 있기 때문에 더욱 정확한 대사물질의 정보를 얻을 수 있다. 본 연구에서는 개 (canine)의 소뇌 (cerebellum)조직으로부터 대사물질을 추출하고 고자장 핵자기공명분광법으로 대사물질의 절대농도를 획득 하고자 하였다. 생체 대사물질의 절대농도를 획득하기 위하여 대표적인 대사물질(i.e., NAA, Cr, Cho, Ins, Lac, GABA, Glu, Gln, Tau Ala)의 팬톰을 제작하여 그 스펙트럼을 확보하였고, 개의 소뇌 부위를 적출하여 methanol-chloroform water extraction (M/C water extraction) 방법으로 대사물질만을 추출한 후 자기공명분광법을 수행하였다. 필터링 (filtering)의 효과를 평가하기 위하여 샘플 제작 시 추출물을 필터링한 그룹과 필터링하지 않은 그룹으로 분류하여 실험을 수행하였다. 팬텀 물질과 추출물은 90% D2O 수용액으로 만든 후 5mm NMR 튜브에 담아 실험하였다. 실험 결과 조직 추출물을 필터링하는 것이 신호대잡음비(signal to noise ratio: SNR, S/N)를 향상시키는 데 기여하는 것을 확인하였다. 또한 개의 소뇌 대사물질의 절대농도는 사람보다는 쥐 (rat)의 뇌 대사물질 절대농도와 더 비슷한 것을 확인할 수 있었다. 본 연구 결과를 토대로 고자장 핵자기공명분광법을 이용한 in vitro 실험은 뇌조직 내 대사물질의 절대농도를 측정하고 기본적인 지표를 확보하는데 매우 정확하고 정량적인 방법이 될 수 있을 것으로 사료된다.

• 대한한방내과학회지
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• 제43권6호
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• pp.1105-1121
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• 2022

Objectives: The purpose of this study is to explore the effectiveness and safety of frequently used clinical herbal medicines (Yijung-tang [Lizhong-tang, LJT], Pyeongwi-san [Pingwei-san, PWS], and Shihosogan-tang [Chaihu Shugan-tang, SST]) in patients with functional dyspepsia (FD) when administered according to herbal medicine and Korean medicine pattern identification. The results of this study will be used to standardize the diagnostic instrument used in Korean medicine and to investigate biomarkers of Korean medicine pattern identification. Methods: This study will be a randomized, assessor-blind, 3-arm, parallel, open-label, multi-center clinical trial. A total of 300 FD participants will be recruited from 3 Korean medical hospitals and assigned to the LJT (n=100), PWS (n=100), and SST (n=100) groups according to FD pattern identification. The patients will take the medication for 8 weeks, 3 times a day, before or between meals. The primary outcome will be total dyspepsia symptom (TDS) and the secondary outcomes will be adequate relief (AR) for dyspepsia, overall treatment effect (OTE), visual analogue scale (VAS), functional dyspepsia-related quality of life (FD-QoL), gastrointestinal symptom score (GIS), and pattern identification questionnaires. For the exploratory outcomes, we will analyze blood and fecal metabolome profiles, microbiota from fecal and saliva samples, single nucleotide polymorphism (SNP), and results of Korean medicine diagnosis device measurements (heart rate variability, and tongue, pulse, and abdominal diagnosis). Conclusions: The results of this study will prove objectivity for Korean medicine pattern identifications, and the effectiveness and safety of herbal medicines for the population with FD.