Comparative metabolomic analysis of corticosterone-induced changes in embryonic mouse hypothalamic cells using gas chromatography coupled with triple-quadrupole tandem mass spectrometry
Title
Comparative metabolomic analysis of corticosterone-induced changes in embryonic mouse hypothalamic cells using gas chromatography coupled with triple-quadrupole tandem mass spectrometry
Alternative Title
GC-QqQ-MS/MS를 사용한 쥐 배아 시상 하부 세포에서의 코르티코스테론 유도 대사체 변화 분석
I. Introduction 1 II. Materials and methods 6 1. Cell culture 6 2. Cell viability assay 6 3. Real time-PCR 7 4. Metabolomics analysis by GC-QqQ-MS/MS 8 4.1 Glycolysis, TCA cycle, and PPP intermediates analysis 8 4.2 Amino acids analysis 9 4.3 Free fatty acids analysis 10 5. ATP assay 11 III. Results 12 1. Corticosterone has cytotoxicity in embryonic mouse hypothalamic cells in a dose-dependent manner 12 2. Corticosterone increases mRNA expression of Agrp, Npy, and Pomc in embryonic mouse hypothalamic cells 12 3. Corticosterone changes metabolites in different ways depending on the cell type 14 3.1 Corticosterone increases glucose metabolism intermediates and decreases amino acids and fatty acids in N41 cells 14 3.2 Corticosterone decreases glucose metabolism intermediates and increases fatty acids in N43/5 cells 16 4. Corticosterone decreases intracellular ATP level and regulates mRNA expression related to fatty acid metabolism in N41 cells 16 5. Pharmacological inhibition of GPAT blocks the increase of Npy and Gpat mRNA expression induced by corticosterone in N41 cells 18 6. Corticosterone decreases intracellular ATP level and increases mRNA expression of Gpat in N43/5 cells 18 7. Pharmacological inhibition of GPAT blocks increase of Pomc mRNA expression induced by corticosterone in N43/5 cells 21 IV. Discussion 24 V. Conclusion 27 VI. References 28