An In Vitro Model to Investigate the Role of Helicobacter pylori in Type 2 Diabetes, Obesity, Alzheimer’s Disease and Cardiometabolic Disease
Abstract
:1. Introduction
2. Results
2.1. NMR-Based Metabolomics Analysis
2.2. Helicobacter Pylori Alters the Amino Acids Metabolism
2.3. MKN-28 Cells Uptake BCAA from Culture Medium
2.4. Helicobacter Pylori Induces Inflammation and Oxidative Stress
3. Discussion
4. Materials and Methods
4.1. Helicobacter Pylori
4.2. Cell Culture Conditions
4.3. Cell Culture for Metabolomics Analysis
4.4. Metabolites Extraction and NMR Samples Preparation
4.5. NMR Spectroscopy
4.6. RNA Extraction and Real-Time PCR
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
Hp | Helicobacter pylori |
NMR | Nuclear Magnetic Resonance |
Hpcf | Hp cell filtrate |
BCAA | Branched chain amino acids |
mTORC1 | mammalian Target Of Rapamycin Complex 1 |
TLRs | Tool like receptors |
NF-AT | nuclear translocation of the T transcription factor |
T2D | Type 2 diabetes |
OB | Obesity |
AD | Alzheimer’s disease |
CMD | Cardiometabolic disease |
PCA | Principal Component Analysis |
OPLS- | Orthogonal partial least squares discriminant analysis |
AAA | Aromatic amino acids |
ROS | Reactive oxygen species |
LAT1 | Large neutral amino acid transporter |
BCAT | Branched chain amino acid transaminase |
KLF15 | Kruppel-like factor 15 |
FBS | Fetal Bovine Serum |
BHI | Brain Heart Infusion medium |
PBS | Phosphate-Buffered Saline |
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Pathway Name | Gene ID | Gene Name | Fold Regulation 1 h | Fold Regulation 2 h |
---|---|---|---|---|
mTOR signaling pathway | RPTOR | Regulatory associated protein of mTOR complex 1 | −1.42 | 286.04 |
MLST8 | mTOR associated protein, LST8 homolog (S. cerevisiae) | −1.42 | 398.95 | |
AKT1 | V-akt murine thymoma viral oncogene homolog 1 | −1.42 | 50.13 | |
AKT2 | V-akt murine thymoma viral oncogene homolog 2 | −1.42 | 504.97 | |
INSR | Insulin receptor | −1.42 | 257.79 | |
IRS1 | Insulin receptor substrate 1 | −1.42 | 278.22 | |
PLD1 | Phospholipase D1, phosphatidylcholine-specific | −6.31 | 130.70 | |
RPS6KA2 | Ribosomal protein S6 kinase, 90 kDa, polypeptide 2 | −1.24 | 3.37 | |
PDPK1 | 3-phosphoinositide dependent protein kinase-1 | −1.53 | 28.25 | |
PIK3CB | Phosphoinositide-3-kinase, catalytic, beta polypeptide | −1.42 | 16.34 | |
PIK3CD | Phosphoinositide-3-kinase, catalytic, delta polypeptide | 3.37 | 184.83 | |
PIK3CG | Phosphoinositide-3-kinase, catalytic, gamma polypeptide | −1.42 | 215.28 | |
CHUK | Conserved helix-loop-helix ubiquitous kinase | −4.08 | 181.03 | |
EIF4E | Eukaryotic translation initiation factor 4E | −1.42 | 922.92 | |
HIF1A | Hypoxia inducible factor 1, alpha subunit | 192.93 | 955.47 | |
Inflammatory pathway | CXCL8 | Interleukin 8 | −3.29 | 2.96 |
IL-6 | Interleukin 6 | 14.45 | 114.56 | |
TLR2 | Toll-like receptor 2 | 58 | 72.18 | |
TLR9 | Toll-like receptor 9 | 3.29 | 134.55 | |
TNF | Tumor necrosis factor | 12.9 | 154.26 | |
Oxidative stress pathway | ATOX1 | ATX1 antioxidant protein 1 homolog (yeast) | 3.57 | 37.69 |
GPX2 | Glutathione peroxidase 2 (gastrointestinal) | 3.57 | 37.69 | |
GPX4 | Glutathione peroxidase 4 (gastrointestinal) | 3.57 | 37.69 | |
GSS | Glutathione synthetase | 3.57 | 9.54 | |
NOX5 | NADPH oxidase. EF-hand calcium binding domain 5 | 3.57 | 7.54 | |
SOD1 | Superoxide dismutase 1. soluble | −28.68 | −9.67 | |
SOD2 | Superoxide dismutase 2. mitochondrial | 3.96 | 4.04 |
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Cuomo, P.; Papaianni, M.; Sansone, C.; Iannelli, A.; Iannelli, D.; Medaglia, C.; Paris, D.; Motta, A.; Capparelli, R. An In Vitro Model to Investigate the Role of Helicobacter pylori in Type 2 Diabetes, Obesity, Alzheimer’s Disease and Cardiometabolic Disease. Int. J. Mol. Sci. 2020, 21, 8369. https://doi.org/10.3390/ijms21218369
Cuomo P, Papaianni M, Sansone C, Iannelli A, Iannelli D, Medaglia C, Paris D, Motta A, Capparelli R. An In Vitro Model to Investigate the Role of Helicobacter pylori in Type 2 Diabetes, Obesity, Alzheimer’s Disease and Cardiometabolic Disease. International Journal of Molecular Sciences. 2020; 21(21):8369. https://doi.org/10.3390/ijms21218369
Chicago/Turabian StyleCuomo, Paola, Marina Papaianni, Clementina Sansone, Antonio Iannelli, Domenico Iannelli, Chiara Medaglia, Debora Paris, Andrea Motta, and Rosanna Capparelli. 2020. "An In Vitro Model to Investigate the Role of Helicobacter pylori in Type 2 Diabetes, Obesity, Alzheimer’s Disease and Cardiometabolic Disease" International Journal of Molecular Sciences 21, no. 21: 8369. https://doi.org/10.3390/ijms21218369