Metabolomic study of possible anti-osteoporotic compounds with the aid of nmr spectroscopy and mass spectrometry

Postgraduate Thesis uoadl:1318915 322 Read counter

Unit:
Κατεύθυνση Φαρμακευτική Ανάλυση - Έλεγχος ποιότητας
Library of the School of Science
Deposit date:
2016-04-19
Year:
2016
Author:
Μπληζιώτης Νικόλαος-Γεώργιος
Supervisors info:
Εμμανουηλ Μικρός Καθηγητής, Ευάγγελος Γκίκας Επίκ. Καθηγητής
Original Title:
Μεταβολομική μελέτη πιθανών αντιοστεοπορωτικών μορίων με την βοήθεια φασματοσκοπίας nmr και φασματομετρίας μάζας
Languages:
Greek
Translated title:
Metabolomic study of possible anti-osteoporotic compounds with the aid of nmr spectroscopy and mass spectrometry
Summary:
Osteoblasts are the major cellular component of bone tissue. Their biological
functions include regulation of osteoclast formation and function,
hematopoietic stem cell homeostasis, various endocrine functions and
ossification. Compounds that promote osteoblast differentiation can be used for
the treatment or prevention of osteoporosis. The aim of the present study was
to evaluate the effect of two active compounds already in use for the treatment
of osteoporosis, i.e. estradiol (E) and raloxifene (R) and two natural
compounds i.e. kaempferol (K) and oleuropein (O) on osteoblast differentiation
through metabolic profiling of MC3T3 cells.
Cells were cultured in plates, and cell pellets were extracted using a three
solvent protocol (MeOH-CHCl3-H2O). Aqueous extracts were analyzed using NMR
spectroscopy (Bruker Avance III 600 MHz) and UPLC-HRMS (Orbitrap Discovery).
Multivariate analysis of high resolution NMR and MS (positive and negative
ionization) data was performed by PCA, PLS-DA and OPLS, while the putative
biomarkers were determined on the basis of the VIP lists, the S-plots and
student’s t-tests for each statistical differentiation.
Results showed differences in the metabolome of E cultured cells compared to R,
as well as O, K compared to all other groups, while groups O and K showed no
distinction. E caused a spike in the concentration of metabolites glutamate,
aspartate, taurine, myo-inositol, creatine, creatine phosphate, methionine,
succinate and threonine. R raised the levels of threonine, tyrosine, valine,
phenylalanine, creatine and taurine. On the other hand, O and K gave rise to
high levels of CTP and ADP, and low levels of ATP, AMP and UDP, an effect
possibly attributed to enhancement of CTP synthase activity. Oleuropein and
Kaempferol also demonstrated high levels of threonine, aspartate,
phenylalanine, valine and tyrosine.
Given that metabolites raised by O and K have been correlated with beneficial
activity on bone, we conclude that O and K could well be an effective
alternative for osteoporosis treatment, but more evidence is needed especially
for the action of these compounds on CTP synthase. Finally, threonine appears
to be the only metabolite raised by all compounds studied, compared to the
controls (DF).
Keywords:
Osteoblast, Metabolomics, Nmr spectroscopy, Mass spectrometry, Threonine
Index:
Yes
Number of index pages:
2
Contains images:
Yes
Number of references:
70
Number of pages:
111
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