XIV
h
International Conference on Molecular Spectroscopy, Białka Tatrzańska 2017
236
T2: P–21
Alkali metal salts of 5-O-caffeoylquinic acid (chlorogenic acid):
spectroscopic, thermogravimetric and biological studies
Monika Kalinowska
1
, Ewelina Bajko
2
, and Włodzimierz Lewandowski
1
1
Division of Chemistry, Bialystok University of Technology, Wiejska 45E Street, 15-351 Bialystok,
Poland, m.kalinowska@pb.edu.pl
2
Faculty of Forestry, Bialystok University of Technology, Pilsudskiego 1A Street,, 17-200 Hajnówka,
Poland
Chlorogenic acids are mono-, di-, tri- or tetra esters of one or more cinnamic acids and
quinic acid, sometimes with an aliphatic acid replacing a cinnamic acid residue. 5-O-
Caffeoylquinic acid (5-CQA, common name: chlorogenic acid) is one of the major chlorogenic
acids present in plant extracts with high biological importance. The synthesis of alkali metal
salts of chlorogenic acid is intended to obtain new compounds with increased solubility in water
medium, lipophilicity, and different biological activity compared to ligand. The molecular
structure of alkali metal chlorogenates was described by means of FT-IR, FT-Raman, UV/VIS,
1H and
13
C NMR spectroscopy. The elemental and thermogravimetric analysis were used in
order to establish the composition of chlorogenates.. The DPPH∙ and FRAP assays were used to
preliminary estimation of the antioxidant properties of synthesized compounds and ligand. The
antioxidant capacity was express by EC
50
parameters (concentration required to obtain a 50%
antioxidant effect).
Keywords: chlorogenic acid; phenolic compounds; antioxidant, spectroscopy
Acknowledgment
This work was funded by the National Science Centre (Poland) on the basis of the decision number
DEC2013/11/D/NZ9/02774
References
[1]
E. Bajko, M. Kalinowska, P. Borowski, L Siergiejczyk, W. Lewandowski, LWT - Food Sci. T. 65
(2016) 471.
[2] M. Kalinowska, A. Witkowska, H. Lewandowska-Siwkiewicz, W. Lewandowski, Plant Physiol.
Bioch. 84 (2014) 169.
XIV
h
International Conference on Molecular Spectroscopy, Białka Tatrzańska 2017
237
T2: P–22
Raman spectroscopy investigation of non-alcoholic fatty liver disease
at single cell level
Ewelina Szafraniec
1
, Bożena Kukla
1
, Edyta Kuś
2
,
Stefan Chlopicki
3
, and Malgorzata Baranska
1,2
1
Faculty of Chemistry,Jagiellonian University, Ingardena 3, Krakow, Poland,
e-mail: baranska@chemia.uj.edu.pl
2
Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego
14, Krakow, Poland
3
Chair
of
Pharmacology,
Jagiellonian
University
Medical
College,
Grzegorzecka
16,
Krakow,
Poland.
Non-alcoholic fatty liver disease (NAFLD) is one of the manifestations of metabolic syndrome,
and its worldwide prevalence is continually increasing. NAFLD is manifested primarily by an
increased accumulation of lipids within hepatic tissue and is known to be strongly associated with
insulin resistance, obesity, hypertension and atherosclerosis.1,2 NAFLD pathogenesis involves an
alteration of a cross-talk between various cells (hepatocytes, liver sinusoidal endothelial cells (LSEC),
hepatic stellate cells (HSC)) in liver tissue and involves changes in their phenotype. In a healthy liver,
fenestrated LSECs without a basement membrane facilitate transport of macromolecules between
blood and hepatocytes, and maintains liver homeostasis by balanced production of mediators.
Hepatocytes possess microvilli while quiescent HSC stores vitamin A . Paracrine action of
hepatocytes and HSC maintains fenestrated phenotype of LSECs by vascular endothelial growth
factor (VEGF) production. In NAFLD liver, LSECs porosity is reduced with presence of basement
membrane. An activated HSC loses vitamin A and changed and changes phenotype towards
myofibroblasts. Fat-loaded hepatocytes lose the hepatocytes microvilli. In addition, the cellular
signaling is shifted towards proinflamatory and profibrogenic pathway.3 Those alteration in cellular
cross-talk, however do not explain entirely the pathogenetic mechanism of NAFLD, which is still not
fully understood yet. NAFLD ranges from simple steatosis to non-alcoholic steatohepatitis (NASH),
which is a more severe form, and can lead to fibrosis, cirrhosis, and eventually to hepatocellular
carcinoma. Therefore, better understanding of NAFLD pathogenesis, early diagnosis and
management of NAFLD are crucial for improving the patient prognosis.
Raman imaging possesses a number of advantages, including minimal sample preparation,
nondestructivity together with the possibility to obtain information of overall biochemical
composition of the sample. Raman spectroscopy provides information on the structure of vibrational
levels related to the bonds in the molecules, as it is based on the study of transitions between
vibrational levels of molecules by their interaction with the incident radiation. Analysis of Raman
spectrum enables an identification of single components of the sample, and together with digital
imaging it is possible to obtain information of spatial distribution of chosen components. High spatial
resolution attainable by Raman imaging enables investigations of single cells at the subcellular level4.
In addition, combining Raman spectroscopic imaging with chemometric methods (k-means cluster
analysis, principal component analysis) allows for analysis of different cellular compartments.
Here we present a unique approach based on a protocol of cell isolation from murine liver
followed by Raman imaging of single cell at subcellular level. Such approach was implemented to the
study changes in biochemical content of freshly isolated primary LSECs and hepatocytes in a murine
model of NAFLD at different stages of disease progression.
Keywords: Raman imaging; LSEC; hepatocytes; NAFLD
Acknowledgment
This work was supported by National Science Centre, grant Symfonia No. No DEC-2015/16/W/NZ4/00070.
References
[1] M. Miyao, et. al., Laboratory Investigation 00 (2015) 1.
[2] C. Michiels, J. Cell. Physiol., 196 (2003) 430.
[3] E. Maslak, A. Gregorius, S. Chlopicki, Pharmacological Reports 67 (2015) 689.
[4] K. Galler. Integr. Biol. 6 (2014) 946.
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