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Proteomic Approach Reveals Inhibition of Glycolysis and
Mevalonate Pathway by Oxysterol Treatment in Liver.

1
Wan-Seog Shim , Chanhee Lee , Jong Hyuk Yoon and Parkyong Song 1*
2
1
1. Department of Convergence Medicine, Pusan National University School of Medicine, Yangsan, 50612, Republic of Korea
2. Neurodegenerative Disease Research Group, Korea Brain Research Institute, Daegu, 41068, Republic of Korea
Abstract Results Results (continued)

Oxysterols are oxygenated derivatives of
cholesterol that are intermediates or Proteomics analysis Compound A can reduce each
even end products in cholesterol Proteomics analysis pathway regulatory mRNA
excretion pathways. As an oxysterol, expression.
compound A is known to regulate Hydroxymethylglutaryl-CoA synthase Lipogenesis pathway
Isoform M1 of Pyruvate kinase PKM
endothelial inflammation and cancer Phosphoglycerate kinase 2 1.5 Control
Acyl-CoA (8-3)-desaturase
progression. However, few studies are Farnesyl pyrophosphate synthase 1ug/mL Compound A
available how compound A affects Fatty acid synthase 2.5ug/mL Compound A
cellular proteomic profiles and metabolic -8 -6 -4 -2 0 1.0 ns
Log 2 Folds
pathway in liver. To analyze systemic (Con vs Comp A) ✱✱✱✱ ✱✱✱ ✱✱
effects of compound A in liver, we Relative amount of protein (regulatory Relative mRNA expression ✱✱✱✱ ✱✱✱✱ ✱✱✱✱
performed liquid chromatography- lipogenesis, glycolysis and mevalonate 0.5 ✱✱✱✱ ✱✱✱✱ ✱✱✱✱ ✱✱✱✱ ✱✱✱✱
tandem mass spectrometry (LC-MS/MS) pathway) is decreased by Compound A ✱✱✱✱
using normal liver cell line. KEGG compared to Control.
pathway analysis proved that some
metabolic pathways such as cholesterol 0.0 FASN ACCa ACCb SREBP1c SREBP2 SREBP1a Elovl6
biosynthesis and glycolysis were No changes in AML12 cell Compound A 24h treat in AML12 cell
significantly reduced by compound A viability upon Compound A Mevalonate pathway
treatment. Additionally, transcripts treatment 1.5 Control
analysis further supported that the 1ug/mL Compound A
2.5ug/mL Compound A
compound A inhibited gene expression,
which related to mevalonate, lipogenesis, 1.2 WST-1 Assay 1.0 ✱✱
and glycolysis pathway in mouse 1.0 Relative mRNA expression
hepatic cell line. Because accumulation
of cholesterol and lipid promotes hepatic 0.8 0.5 ✱✱✱✱ ✱✱✱✱
fatty liver disease, and increased Cell viability 0.6 ✱✱✱✱ ✱✱✱✱ ✱✱✱✱ ✱✱✱✱ ✱✱✱✱ ✱✱✱✱
glycolysis contributes to triacylglycerol ✱✱✱✱ ✱✱✱✱ ✱✱✱✱ ✱✱✱✱
maturation, the suppressive effects of 0.4 0.0 HMGS HMGR MVK MVD IDI1 FPPS
compound a in hepatic lipid synthetic 0.2 Compound A 24h treat in AML12 cell
pathway may provide protective effects
against fatty liver development. 0.0 Control 0.25ug/mL 0.5ug/mL 1ug/mL 2.5ug/mL 5ug/mL Glycolysis pathway
Compound A 24h treat in AML12 cell 1.5 Control
Introduction Nt 0.25ug/mL 0.5ug/mL ns 1ug/mL Compound A
2.5ug/mL Compound A
Relative mRNA expression 1.0 n.s ✱ ✱✱✱✱ ✱✱✱✱ n.s ✱

0.5

1ug/mL 2.5ug/mL 5ug/mL ✱✱✱✱ ✱✱✱✱
0.0
PGK PKM1 ALDOB PKLR PFKL
Compound A 24h treat in AML12 cell
Compound A can significantly decrease
transcript levels, involved in lipogenesis,
mevalonate, and glycolysis in liver.
A lot of metabolisms such as
lipogenesis, catabolic, mevalonate,
and glycolysis, which are essential Through WST-1 assay and FACs
metabolic pathways for human occur analysis to measure cell proliferation or
in the liver. Although changes in apoptotic feature, we conclude that
hepatic metabolism by cholesterol are Compound A has no effects on cell
characterized, potential effects of viability.
oxysterol toward liver have not been
fully established yet.
Thus, we show how Compound A,
oxysterol, affect to liver metabolism.
Conclusion

✓ Compound A has no effects on cell survivals.

✓ Compound A inhibits Lipogenesis, Mevalonate, and Glycolysis pathway.

✓ Therefore, we suggest that Compound A has positive effects on Liver metabolism.
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