Dose-dependent target diversion of Danhong injection on the Glu-GLT-1/Gly-GlyR alpha dynamic balance module of cerebral ischemia

Function-oriented modular structure analysis is a great challenge in module-based pharmacological studies. A strategy to uncover target-target interaction (TTI) and dynamic balance regularity (DBR) was established to discover the structural factors influencing modular functions and explore the mechanism of Danhong injection (DHI) in treating cerebral ischemia. The dose-related metabolic features of DHI intervention were investigated using metabolomics and modular pharmacology. The findings indicated that Glu/Gly was a biomarker and GluGLT-1/Gly-GlyR alpha was the core unit regulated by DHI. Gly and Glu displayed opposite patterns and functional roles, representing intra-modular balance. GlyR alpha was identified as the upstream target and GLT-1 as the downstream target by inhibiting or activating GlyR alpha, indicating that DHI has two dose-dependent regulatory modes. GlyR alpha was the major target at low doses, while GLT-1 was activated as the dominant target as doses accumulated. Our study reveals that target-target interaction and dynamic balance regularity are the key factors influencing modular functions, which is a promising breakthrough for module-based pharmacological studies.