Discrimination geographical origin between cabbages from Korea and China by
1H NMR-Based Metabolomics
Jahan Kim1, Young-ae Jung1, Kwang-Sik Lee2, Geum-Sook Hwang1,3*
1
Korea Basic Science Institute, Seoul 136-713, Republic of Korea,
Division of Earth and Environmental Science, Korea Basic Science Institute,
Deajeon 305-333, Republic of Korea
3
Graduate School of Analytical Science and Technology, Chungnam University,
2
Daejeon, 305-764, Republic of Korea
Metabolomic analysis was performed to determine the geographical origin of
cabbage, which is very popular food in East Asia, by NMR with multivariate statistical
analysis technique. Pattern recognition method, such as principal component analysis
(PCA) revealed clear discriminations between two kinds of cabbages (Chunjung,
Chunmyung) from Korea and China. The major metabolites that contributed to the
discrimination between cabbages originated from different region were 4-aminobutyrate
(GABA), lactate, leucine, isoleucine, phenylacetate, phenylalanine, sucrose, tyrosine,
and valine.
In particular, GABA, leucine, phenylacetate, phenylalanine have known as the
most significant metabolites to differentiate the growing area. The two kinds of
cabbages showed a similar metabolic pattern. This result suggests that the major
metabolites that are significantly different between cabbages from two geographical
origins were influenced by the environmental conditions such as temperature, rainfall,
latitude, humidity and soil type of each country. Our study demonstrated that 1H NMR
based on metabolomics coupled with multivariate statistics can be applicable to
discriminate the geographical origin of cabbages.
Metabolomics study on alcoholic fatty liver in Zebrafish using 1H NMR and
multivariate statistical analysis
Zi-Hey Jang1,2, Chul-Hee Kim2 and Geum-sook Hwang1,2*
1
2
Korea Basic Science Institute, Seoul 136-713, Republic of Korea,
Graduate School of Analytical Science and Technology, Chungnam University,
Daejeon, 305-764, Republic of Korea
Zebrafish that has been a prominent model in developmental biology is now
also gaining popularity as a model for disease study and drug discovery. In addition, this
fish is highly suitable for whole organism assessment of changes in global expression
due to chemicals or drugs. Metabolomics is a promising new approach for
understanding of metabolic perturbations in drug toxicity and disease status. Here, we
investigated the metabolic changes in alcoholic fatty liver from Zebrafish model using
1
H NMR based metabolic profiling.
In this study, several metabolites that differentiate the alcoholic fatty liver group
from the control group were thoroughly characterized and the metabolic changes
between alcoholic fatty liver and control groups were investigated by 1H NMR
spectroscopy coupled with multivariate analysis. This analytical approach was used to
generate a molecular fingerprint of alcoholic liver samples from Zebrafish, and then
pattern recognition technique was applied to identity molecular signatures associated
with the alcoholic liver. Such metabolic signatures could provide diagnostic markers for
a disease state and mechanistic information on cellular perturbations and pathways.
Metabolic profiling of renal cell in rats with chronic renal failure
by 1H NMR and LC MS
Yoo-Jeong Yoon1, 2, Jin-sup Kim1,3 , Hyo-Jung Choi 4, Tae-Hwan Kwon 4, GeumSook Hwang1, 3*
1
Korea Basic Science Institute, Seoul 136-713, Republic of Korea,
Department of chemistry, Korea University Graduate School, Seoul 136-701
3
, Graduate School of Analytical Science and Technology, Chungnam University,
2
4
Daejeon, 305-764, Republic of Korea,
Department of Biochemistry and Cell Biology School of Medicine, Kyungpook
National University Dongin-dong 101, Taegu 700-42, Republic of ,Korea
Metabolomics is the measurement of metabolites within an organism, thereby
providing valuable insight into the metabolism of cells. In this research, IMCD cells (rat
inner medullary collecting duct) with chronic renal failure were characterized using 1HNMR spectroscopy and Liquid chromatography- Mass spectrometer followed by
principal component analysis. We have devised an analytical approach that combines
high resolution 1H NMR and LC MS to provide both a large amount of structural
information and quantification of a broad spectrum of compounds. This combined
approach also simplified sample preparation and reduces the analysis time, making it
well-suited for a convenient screening of metabolites. Major endogenous metabolites
for IMCD cell contained products of amino acids and organic osmolytes (e.g., sorbitol,
betaine, myo-inositol, taurine, and glycerophosphocholine). Many metabolites revealed
changes in the profiles of cell metabolites between control and low osmolality group
which were influenced by chronic renal failure. Metabolomics of kidney cell based on
1H-NMR spectroscopy and LC MS could provide insight into the effects of osmolality.
NMR-based metabolomics approach for characterizing the effect of environmental
stressors on marine mussels (Mytilus)
Jong-Chul Park1,2, Geum-Sook Hwang1, 2*
1
2
Korea Basic Science Institute, Seoul 136-713, Republic of Korea,
Graduate School of Analytical Science and Technology, Chungnam University,
Daejeon, 305-764, Republic of Korea
Marine mussels (Mytilus) have been widely used as biomonitors because they
can be subjected to environmental stresses and pollutants in the natural environment. In
this study, we were aimed at investigating the effect of the environmental stressors on
marine mussels. 1H NMR spectroscopy coupled with multivariate statistical analyses
was applied to metabolomic analyses of mantle tissue extracts obtained from two
different regions, Onsan Bay and Dokdo. Metabolite profiling was performed to
investigate the metabolite differences of marine mussels from different origins and to
identify the significant metabolites associated with environmental stressors. In addition,
heavy metal concentrations in marine mussels obtained from two different regions were
analyzed by using ICP-MS. Mussels from Onsan Bay showed higher levels of Pb, Zn,
Cu, and Mn compared to those from Dokdo. Principal component analysis (PCA)
showed significant separation between marine mussels originating from two regions,
indicating distinctive metabolic differences between mussels from Onsan Bay and
Dokdo. Mussels from Onsan Bay were characterized by increased levels of metabolites
such as amino acids, arginine, betain, glutamine, and taurine, exhibiting metabolic
perturbation in osmoregulation, energetic metabolism, and neurotoxicity. This study
suggest that NMR-based metabolomics is an efficient method to distinguish
fingerprinting difference between mussels originating from different environment and is
suitable for elucidating possible biological pathway perturbed by effects of
environmental stressors.
Metabolomic Approach to Understand the Regulation of Cancer Cell Metastasis
under Glucose Deprivation
Ju-Eun Lee1,2, Ju-Ae Kim1,2, Do-Hyun Ryu2 and Geum-Sook Hwang1,3*
1
Korea Basic Science Institute, Seoul 136-713, Republic of Korea,
²Sungkyunkwan Univ. Natural Sciences Campus, Cheoncheon-dong, Jangan-gu,
Suwon-si, Gyeonggi-do 440-746, Republic of Korea,
3
Graduate School of Analytical Science and Technology, Chungnam University,
Daejeon, 305-764, Republic of Korea
Most cancer cells rely on aerobic glycolysis to generate energy and proliferate
for cellular processes, a phenomenon termed “the Warburg effect”. However, this
clinical
assumption that glucose is supplied unlimitedly does not apply
manifestation
manifestation in human. After cancer cells stop their proliferation under
glucose deprivation, they start to metastasize and implement
strategy
clinical
a
a survival
strategy. Though there have been many studies which describe this
little
is
know about
phenomena,
little
is
known about the
metabolic pathway.
In this study, metabolic profiling using 1H nuclear magnetic resonance(NMR)
spectroscopy coupled with multivariate statistics was used to study the metabolism in
metastatic cancer cell line MDA-MB-231. Metabolic difference between parent cell
grown in sufficient glucose medium and selected cells survived from glucose
deprivation was investigated to reveal the regulation metabolism of anti-apoptosis and
metastasis in cancer cells.
The metabolites that were significantly different between the two cell
conditions include amino acids, energy metabolites, glycolytic end-products, and
osmolytes. Level of metabolites such as amino acids that consist of building blocks in
cell and TCA cycle intermediate metabolites such as succinate were decreased in
selected cell compared to parent cell. On the other hand, osmolytes such as myo-inositol
and glycero-phosphocholine, which is responsible to regulate homeostasis and volume
of cell, were considerably increased in selected cell. This result demonstrates
proliferation is reduced and metastasis is initiated in cancer cell under glucose
deprivation condition. In conclusion, NMR-based metabolic profiling of cancer cell can
be used to evaluate the changes in various cell conditions to elucidate biological process.
1H-NMR-Based
Metabolomics Study of Cerebral Infarction
Jee-Youn Jung1,2, Geum-Sook Hwang1,3 *
1
Seoul center, Korea Basic Science Institute, Seoul, 136-701, Republic of Korea,
2
Department of physiology, College of Oriental Medicine, Wonkwnag University,
Iksan, 540-749, Republic of Korea,
3
Graduate School of Analytical Science and Technology, Chungnam University,
Daejeon, 305-764, Republic of Korea
Stroke is one of the leading causes of adult disability and death in developing
countries. However, early diagnosis is difficult and no reliable biomarker is currently
available. Thus, we applied a 1H-NMR metabolomics approach to investigate the altered
metabolic pattern in plasma and urine from patients with cerebral infarctions and sought
to identify metabolic biomarkers associated with stroke.
Metabolic profiles of plasma and urine from patients with cerebral infarctions,
especially small vessel occlusion (SVO), were investigated using 1H-NMR spectroscopy,
coupled with multivariate statistical analysis, such as principal components analysis
(PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA).
Multivariate statistical analysis showed a significant separation between patients
and healthy individuals. The plasma of stroke patients was characterized by the
increased excretion of lactate, pyruvate, glycolate, formate, and by the decreased
excretion of glutamine and methanol; the urine of stroke patients was characterized by
decreased levels of citrate, hippurate, and glycine. These metabolites detected from
plasma and urine of patients with cerebral infarction were associated with anaerobic
glycolysis, folic acid deficiency, and hyperhomocysteinemia. Furthermore, the presence
of cerebral infarction in the external validation model was predicted with high accuracy.
These data demonstrate that a metabolomics approach may be useful for the
effective diagnosis of cerebral infarction, and further understanding of stroke
pathogenesis.
1H-NMR-Based
Metabolomics Study on Osteoporosis
with multivariate statistical analysis.
Eun-Jin Kim1,2, Geum-Sook Hwang1,3*
1
2
Korea Basic Science Institute, Seoul 136-713, Republic of Korea,
Department of Biotechnology, Korea University Graduate School, Seoul 136-701,
Republic of Korea,
3
Graduate School of Analytical Science and Technology, Chungnam University,
Daejeon, 305-764, Republic of Korea
Osteoporosis is a common and significant health problem. This disease is
characterized by a deterioration of the skeleton leading to a high incidence of bone
fractures. To research the biomarker of osteoporosis, change of the endogenous
metabolites in serum detected from osteoporotic mouse by ovariectomy. In this study, 8
mice were randomly divided into sham-operated(sham) and ovariectomized(OVX)
group. Serums were collected from both groups, respectively. The metabolic profiles
were investigated using 1H-NMR spectrometry coupled with multivariate statistical
analysis such as principal components analysis (PCA). Multivariate statistical analysis
showed a significant separation between sham and OVX. The serum of OVX was
characterized by the increased level of Alanine, Glutamine, Lactate, Phenylalanine,
Pyruvate, Succinate and Valine, and by the decreased level of Glucose. These
metabolites influenced of separation both groups. This analytical approach, in
combination with chemometrics by multivariate analysis, is expected to offer a
diagnostic method of diseases with high reliability and consistent results.