Optimal
drug proportions and mechanism deciphering of multicomponent drugs are critical for developing novel
therapies to cope with complex diseases, such as
stroke. In the present study, orthogonal experimental design was applied to explore the optimal proportion of the four component herbs in
Huang-Lian-Jie-Du-Decoction (HLJDD) on the treatment of
ischemic stroke. The treatment efficacies and mechanisms were assessed using global and
amino acids (AAs) targeted metabolomics, as well as correlation network analysis. The global NMR metabolomics results revealed that AAs metabolism was significantly perturbed in
middle cerebral artery occlusion rats. The levels of 23 endogenous AAs were then subjected to HPLC-QTOF-MS/MS analysis. These results complemented with neurobehavioral evaluations,
cerebral infarct assessments, biochemical evaluations, histological inspections and immunohistochemistry observations strongly demonstrated that HLJDD with optimal proportion of 6 (Rhizoma coptidis): 4 (Radix scutellariae): 1 (Cortex phellodendr): 3 (Fructus Gardeniae) had the best efficacy on
ischemic stroke, which could be ascribed to its modulation on AA metabolism. This integrated metabolomics approach showed the potential and applicable in deciphering the complex mechanisms of
traditional Chinese medicine formulae on the treatment of complicated diseases, which provided new means to assess the treatment effects of herb combinations and to further development of drugs or
therapies based on these formulae.