ABSTRACT
AIM: To investigate the effects of octreotide on metabolism in the A549 cells treated with lipopo-lysaccharides ( LPS).METHODS: The technologies of gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) were used to test the metabolism of lung A549 cells subject to different treat-ment with LPS and/or octreotide.The results were visualized and checked by chromatogram, and the corresponding intensi-ty data were analyzed by the principal component analysis(PCA)method.The metabolites with different expression and the underlying interaction network were resolved.RESULTS: The metabolism analysis by LC/MS method indicated that there were different expression levels between different treated groups.Further analysis was carried out by orthogonal projections to latent structures-discriminant analysis (OPLS-DA) and the different expressed metabolites were obtained, which were mainly amino acids and phospholipids.By analyzing with GC/MS method and t-test, the different expressed metabolites were mainly organic acid, saccharides and amino acid metabolite.The interaction network diagram was constructed about the response of A549 cells induced by LPS and/or octreotide, including glycolysis/gluconenogenesis, tryptophan metabo-lism, galactose metabolism, urine cycle and citrate cycle.Fourteen key components were found such as serotonin, indole, threonine, serine, glucose, phenylalanine, lactose, fumarate, 4-hydroxyphenyllactate, aspartate, asparagine, putrescine, proline and succinate.CONCLUSION: In octreotide treated LPS-induced A549 cells, the main metabolites are organic acid, saccharides, amino acids and phospholipids.The interaction network is constructed, including 5 metabolic pathways and 14 key components.
ABSTRACT
AIM:To study the morphological changes of cardiac H9c2 cells during the developmental process of fetal rat.METHODS:Embryonic rat heart-derived H9c2 cells were maintained in DMEM supplemented with 10%fetal bovine serum.The H9c2 cells were plated at a density of 6 000 cells/cm and divided into 5 groups:H9c2 cells were trea-ted with 5 mmol/L glucose, 25 mmol/L glucose, 50 mmol/L glucose, Norvasc (25 nmol/L) +25 mmol/L glucose, or Norvasc (25 nmol/L)+50 mmol/L glucose for 48 h.The morphology of H9c2 cells was observed.The cell surface area was measured by Image-Pro Plus 6.1 software.Fluorescence spectrophotometry was used to detect the concentration of in-tracellular calcium ion ( [ Ca2+] i ) in the cardiomyocytes.The concentration of CaN in the cell was measured by ELISA. The mRNA expression of CaNAβ, NFAT3 and β-MHC in the cells was detected by real-time PCR.The protein levels of CaNAβ, NFAT3 and β-MHC in cultural H9c2 cells were detected by Western blot.RESULTS: The mean area of the cells, the mean fluorescence value of [ Ca2+] i and the concentration of CaN in 25 mmol/L glucose group were higher than those in 5 mmol/L glucose group, and those were lower than those in 50 mmol/L glucose group.After treated with Nor-vasc, those results decreased significantly.The expression of CaNAβ, NFAT3 andβ-MHC at mRNA and protein levels in 25 mmol/L glucose group was higher than those in 5 mmol/L glucose group, but was lower than those in 50 mmol/L glu-cose group .The expression of CaNAβ, NFAT3 andβ-MHC at mRNA and protein levels decreased significantly in Norvasc treatment group.CONCLUSION:Ca2+-CaN-NFAT3 signaling pathway is perhaps involved in high glucose-induced H9c2 cardiomyocyte hypertrophy.