Metabonomics study of the therapeutic mechanism of Gynostemma pentaphyllum and atorvastatin for hyperlipidemia in rats

Miao Wang, Fei Wang, Yinan Wang, Xiaonan Ma, Min Zhao, Chunjie Zhao, Miao Wang, Fei Wang, Yinan Wang, Xiaonan Ma, Min Zhao, Chunjie Zhao

Abstract

Gynostemma pentaphyllum (GP) is widely used for the treatment of diseases such as hyperlipidemia, fatty liver and obesity in China, and atorvastatin is broadly used as an anti-hyperlipidemia drug. This research focuses on the plasma and liver metabolites in the following four groups of rats: control, a hyperlipidemia model, a hyperlipidemia model treated with GP and a hyperlipidemia model treated with atorvastatin. Using (1)H-NMR-based metabonomics, we elucidated the therapeutic mechanisms of GP and atorvastatin. Orthogonal Partial Least Squares-Discriminant analysis (OPLS-DA) plotting of the metabolic state and analysis of potential biomarkers in the plasma and liver correlated well with the results of biochemical assays. GP can effectively affect lipid metabolism, and it exerts its anti-hyperlipidemia effect by elevating the level of phosphatidylcholine and decreasing the level of trimethylamine N-oxide (TMAO). In contrast, atorvastatin affects hyperlipidemia mainly during lipid metabolism and protein metabolism in vivo.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Typical 600 MHz 1 H-NMR…
Figure 1. Typical 600 MHz 1H-NMR spectra of rat plasma samples.
1.Lipids (VLDL/LDL) 3. Isoleucine 4. Valine 5.3-Hydroxybutyrate 6. Lactate 7. Alanine 8. Lysine 9. Arginine 12. N-Acetyl glycoproteins 14. Glutamate 16. Acetoacetate 17. Acetone 18. Succinate 19. Pyruvate 20. Glutamine 21. Citrate 22. Glutathione 23. Aspartate 24. Creatine 26. Choline 27. Phosphocholine/GPC 28. TMAO 30. Glucose/aminoacids resonances 34.α-Glucose 35. Glycogen 37. Fumarate 38. Tyrosine.
Figure 2. Typical 600 MHz 1 H-NMR…
Figure 2. Typical 600 MHz 1H-NMR spectra of rat liver samples.
1. Lipids (VLDL/LDL) 2. Leucine 3. Isoleucine 4. Valine 5.3-Hydroxybutyrate 6. Lactate 7. Alanine 8. Lysine 9. Arginine 10. Acetate 11. Proline 12. N-Acetyl glycoproteins 13. O-Acetyl glycoproteins 14. Glutamate 15. Methionine 16. Acetoacetate 17. Acetone 18. Succinate 19. Pyruvate 20. Glutamine 21. Citrate 22. Glutathione 23. Aspartate 24. Creatine 25. Phosphatidylcholine 26. Choline 27. Phosphocholine/GPC 28. TMAO 29. Taurine 30. Glucose/aminoacids resonances 31.myo–Inositol 32. Threonine 33. β-Glucose 34.α-Glucose 35. Glycogen 36. Adenosine/Inosine 37. Fumarate 38. Tyrosine 39. Phenylalanine 40. Histidine.
Figure 3. PR analysis of the 1…
Figure 3. PR analysis of the 1H-NMR spectra of rat plasma.
(A): PCA analysis of the spectra of plasma from normal and hyperlipidemia rats (R2X=0.988, Q2=0.885). (B): Scores plot of the OPLS-DA analysis of the spectra from the plasma of normal and hyperlipidemia rats (R2X=0.925, R2Y=0.874, Q2=0.642). (C): Scores plot of the OPLS-DA analysis of the spectra from the plasma of normal, hyperlipidemia and GP-treated rats (R2X=0.968, R2Y=0.878, Q2=0.538). (D): Scores plot of the OPLS-DA analysis of the spectra from the plasma of normal, hyperlipidemia and Atorvastatin-treated rats (R2X=0.909, R2Y=0.522, Q2=0.328). (E): Loading plot of the OPLS-DA analysis of the spectra from the plasma of normal and hyperlipidemia rats.
Figure 4. PR analysis of 1 H-NMR…
Figure 4. PR analysis of 1H-NMR spectra of rat liver tissues.
(A): PCA analysis of the spectra of liver tissues from normal and hyperlipidemia rats (R2X=0.955, Q2=0.782). (B): Scores plot of the OPLS-DA analysis of the spectra from the liver tissues of normal and hyperlipidemia rats (R2X=0.953, R2Y=0.999, Q2=0.827). (C): Scores plot of the OPLS-DA analysis of the spectra from the liver tissues of normal, hyperlipidemia and GP-treated rats (R2X=0.955, R2Y=0.984, Q2=0.608). (D): Scores plot of the OPLS-DA analysis of the spectra from the liver tissues of normal, hyperlipidemia and Atorvastatin-treated rats (R2X=0.931, R2Y=0.945, Q2=0.544). (E): Loading plot of the OPLS-DA analysis of the spectra from the liver tissues of normal and hyperlipidemia rats.
Figure 5. Summary of the metabolic pathways…
Figure 5. Summary of the metabolic pathways related to the metabolites that changed significantly in the hyperlipidemia model.
“↑” and “↓” indicate that the compound is up- and down-regulated compared with the control group.

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