Abstract
Aim:
To determine how the relative amino acid contents and metabolic pathways regulate the pharmacological phenotypes in rats with cerebral ischemia after treatment with varying doses of DanHong injection (DHI).
Methods:
Adult male rats underwent middle cerebral artery occlusion (MCAO), and were injected with DHI (DH-1: 1 mL/kg; DH-2: 2.5 mL/kg; DH-3: 5 mL/kg, and DH-4: 10 mL/kg, iv) daily for 3 d. The neurological deficit score, body weights and infarct volume were assessed. Serum levels of 20 free amino acids were determined using HPLC, and the values were transformed through the quantitative analysis of the amino acids in the serum metabolic spectrum. Multivariate statistical analysis methods (PCA and PLS-DA) and web-based metabolomics tools (MetPa and MetaboAnalyst) were used to analyze the biological data sets for the amino acids.
Results:
Administration of DHI dose-dependently decreased cerebral infarct volume, and ameliorated neurological deficits. A total of 5, 6, 7 and 7 non-overlapping metabolites were identified in the DH-1, DH-2, DH-3, and DH-4 groups, respectively. Eight metabolites were shared between the DHI groups and the vehicle group. In addition, the serum levels of glutamic acid, aspartic acid and serine increased with increasing DHI dose. A total of 3, 2, 2 and 5 non-overlapping metabolic pathways were identified in the DH-1, DH-2, DH-3 and DH-4 groups, respectively, and glycine, serine, threonine and histidine metabolism were identified as overlapping pathways among the 4 dose groups.
Conclusion:
Overlapping and non-overlapping amino acid metabolites and metabolic pathways are associated with the dose-dependent neuroprotective effect of DHI.
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Acknowledgements
This research was funded by the Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China. Buchang Pharmaceutical provided the drugs used in this study. However, the funder played no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The researchers are all independent from the funder. This study was part of the National Project of New Drug R&D (2011ZX09304).
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Supplementary information is available at website of Acta Pharmacologica Sinica.
Supplementary information
Supplementary Information, Figure S1
The HPLC-fingerprinted from different batches of DanHong injection. (DOC 198 kb)
Supplementary Information, Figure S2
The quality control condition of Danhong injection (DOC 25 kb)
Supplementary Information, Figure S3
Analysis of the metabolic pattern in rat serum by HPLC-FLD. (DOC 396 kb)
Supplementary Information, Figure S4
The changing process of the number and properties of metabolic pathways under different DH doses treatment. (DOC 299 kb)
Supplementary Information, Table S1
The amino acid names matched with some codes shown in Fig. S4 (DOC 29 kb)
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Guo, Zl., Zhu, Y., Su, Xt. et al. DanHong injection dose-dependently varies amino acid metabolites and metabolic pathways in the treatment of rats with cerebral ischemia. Acta Pharmacol Sin 36, 748–757 (2015). https://doi.org/10.1038/aps.2014.167
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DOI: https://doi.org/10.1038/aps.2014.167
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