Serum Metabonomics Reveals Risk Factors in Different Periods of Cerebral Infarction in Humans

Abstract

Studies of key metabolite variations and their biological mechanisms in cerebral infarction (CI) have increased our understanding of the pathophysiology of the disease. However, how metabolite variations in different periods of CI influence these biological processes and whether key metabolites from different periods may better predict disease progression are still unknown. We performed a systematic investigation using the metabonomics method. Various metabolites in different pathways were investigated by serum metabolic profiling of 143 patients diagnosed with CI and 59 healthy controls. Phe-Phe, carnitine C18:1, palmitic acid, cis-8,11,14-eicosatrienoic acid, palmitoleic acid, 1-linoleoyl-rac-glycerol, MAG 18:1, MAG 20:3, phosphoric acid, 5α-dihydrotestosterone, Ca, K, and GGT were the major components in the early period of CI. GCDCA, glycocholate, PC 36:5, LPC 18:2, and PA showed obvious changes in the intermediate time. In contrast, trans-vaccenic acid, linolenic acid, linoleic acid, all-cis-4,7,10,13,16-docosapentaenoic acid, arachidonic acid, DHA, FFA 18:1, FFA 18:2, FFA 18:3, FFA 20:4, FFA 22:6, PC 34:1, PC 36:3, PC 38:4, ALP, and Crea displayed changes in the later time. More importantly, we found that phenylalanine metabolism, medium-chain acylcarnitines, long-chain acylcarnitines, choline, DHEA, LPC 18:0, LPC 18:1, FFA 18:0, FFA 22:4, TG, ALB, IDBIL, and DBIL played vital roles in the development of different periods of CI. Increased phenylacetyl-L-glutamine was detected and may be a biomarker for CI. It was of great significance that we identified key metabolic pathways and risk metabolites in different periods of CI different from those previously reported. Specific data are detailed in the Conclusion section. In addition, we also explored metabolite differences of CI patients complicated with high blood glucose compared with healthy controls. Further work in this area may inform personalized treatment approaches in clinical practice for CI by experimentally elucidating the pathophysiological mechanisms.

Keywords: cerebral infarction; different periods; human serum; key metabolites and pathways; metabonomics.

FIGURE 1

Metabolic profiling analysis of different periods of cerebral infarction in humans. (A) Score plot of the samples using PCA model in positive ions. (B) Score plot of the samples using PCA model in negative ions. (C) RSD (relative standard deviation) distribution of the ions in QC samples. (D) Score plot of samples from healthy controls (N) and attacks within 3 days of cerebral infarction (A) group using PLS-DA model. (E) Score plot of samples from healthy controls (N) and onsets after 3 days but within 5 days of cerebral infarction (B) group using PLS-DA model. (F) Score plot of samples from healthy controls (N) and seizures after 5 days but within 7 days of cerebral infarction (C) group using PLS-DA model. (G) Score plot of samples from healthy controls (N) and attacks after 7 days of cerebral infarction (D) group using PLS-DA model. (H) Score plot of samples from healthy controls (N) and cerebral infarction patients with glycosuria (T) group using PLS-DA model. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20.

FIGURE 2

Differential metabolites found by metabolomics analysis and metabolic pathway analysis. (A–E) It represents the results of metabolic pathway in different treatment groups. The abscissa is $-{\log }_{\text{e}}\left(\text{p}\right)$ , the ordinate is metabolic pathway. (F) It presents the relationship of amino acid pathways in different treatment groups in the Venn diagram. Venn diagrams of samples from healthy controls (N) and cerebral infarction (A–D) group and cerebral infarction and glycosuria (T) group. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20.

FIGURE 3

Changes in carbohydrates and amino acids of cerebral infarction. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20. All data are the mean ±SEM. *p < 0.05 vs. N group; **p < 0.01 vs. N group, two-tailed Mann–Whitney U test.

FIGURE 4

Metabolomics analysis in bile acids of cerebral infarction. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20. All data are the mean ±SEM. **p < 0.01 vs. N group, two-tailed Mann–Whitney U test.

FIGURE 5

Metabolomics analysis in carnitines of cerebral infarction. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20. All data are the mean ±SEM. *p < 0.05 vs. N group; **p < 0.01 vs. N group, two-tailed Mann–Whitney U test.

FIGURE 6

Changes in stearate metabolic pathway analysis of cerebral infarction. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20. All data are the mean ±SEM. *p < 0.05 vs. N group; **p < 0.01 vs. N group, two-tailed Mann–Whitney U test.

FIGURE 7

Changes in cholesterol metabolic pathway of cerebral infarction. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria group, n = 20. All data are the mean ±SEM. *p < 0.05 vs. N group; **p < 0.01 vs. N group; dt, p < 0.05 D group vs. T group; two-tailed Mann–Whitney U test.

FIGURE8

Changes in PC, LPC, and FFA metabolic pathway of cerebral infarction. N, healthy controls, n = 41; A, attacks within 3 days of cerebral infarction, n = 31; B, onsets after 3 days but within 5 days of cerebral infarction, n = 17; C, onsets after 3 days but within 5 days of cerebral infarction, n = 13; D, attacks after 7 days of cerebral infarction, n = 19; T, cerebral infarction patients with glycosuria, n = 20. All data are the mean ±SEM. *p < 0.05 vs. N group; **p < 0.01 vs. N group; bt, p < 0.05 B group vs. T group; dt, p < 0.05 D group vs. T group; two-tailed Mann–Whitney U test.

FIGURE 9

Blood biochemistry examination in various stages of cerebral infarction. N, healthy controls, n = 59; A, attacks within 3 days of cerebral infarction, n = 38; B, onsets after 3 days but within 5 days of cerebral infarction, n = 25; C, onsets after 3 days but within 5 days of cerebral infarction, n = 18; D, attacks after 7 days of cerebral infarction, n = 30; T, cerebral infarction patients with glycosuria, n = 32. All data are the mean ±SEM. *p < 0.05 vs. group; **p < 0.01 vs. group; ***p < 0.01 vs. group; two-tailed Mann–Whitney U test.

FIGURE 10

Graphical abstract, conclusions of our study in different periods of Cerebral Infarction in Human. The red arrows indicate that it is elevated in this kind of metabolites, the blue arrows indicate that it is elevated in this kind of metabolites suffering from cerebral infarction.