Metabolomics coupled with SystemsDock reveal the protective effect and the potential active components of Naozhenning granule against traumatic brain injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Naozhenning granule (NZN), a widely traditional Chinese medicine (TCM) prescription with a long history of clinical, which is mainly used in the treatment of concussion, cerebral post-traumatic syndrome, consists of Di Huang (Radix of Rehmannia glutinosa (Gaertn.) DC.), Dang Gui (Radix of Angelica sinensis (Oliv.) Diels), Chen Pi (Pericarpium of Citrus reticulata Blanco), Dan shen (Radix of Salvia Miltiorrhiza Bunge.), Di Long (Pheretima aspergillum (E. Perrier)), Mu Dan Pi (Cortex of Paeonia suffruticosa Andrews), Suan Zao Ren (Semen of Ziziphus jujuba Mill.), Chuan Xiong (Rhizoma of Ligusticum striatum DC.), Zhu Ru (Phyllostachys nigra (Lodd. Ex Lindl.) Munro), Bai Zi Ren (Semen of Platycladus orientalis (L.) Franco) and Fu Ling (sclerotium of Poria cocos (Schw.)Wolf). AIM OF THE STUDY: This study aimed to unravel the mechanism and material basis of NZN against traumatic brain injury. MATERIALS AND METHODS: In this study, a 1H nuclear magnetic resonance (NMR) based metabolomic approach combined with systemsDock was employed to study the protective effect of NZN against traumatic brain injury using a cerebral concussion rat model. The morris water maze test and biochemical indexes were used to evaluate the efficacy of NZN. RESULTS: The results of morris water maze test suggested NZN can improve the spatial learning and memory of model rats, and the superoxide dismutas (SOD) and malonyldialdehyde (MDA) level indicated that the effect of NZN was related with the regulation of oxidative stress. Multivariate analysis revealed that the effect of NZN was related with regulation of 18 brain metabolites, and the corresponding metabolic pathways were further revealed by MetPA analysis. 13 serum absorbed components were found to hit the targets both related with the metabolic regulation and cerebral trauma through systemsDock-aided molecular docking experiments, and these compounds might be served as the active compounds in NZN against cerebral trauma. CONCLUSION: 1H-NMR based metabolomics and molecular docking provided the insights for the synergistic mechanisms and the potential active compounds of NZN in treating cerebral trauma. However, the bioactive compounds and their synergistic effect need to be further validated.

Rapid characterization of the absorbed constituents in rat serum after oral administration and action mechanism of Naozhenning granule using LC-MS and network pharmacology.

Naozhenning granule is a Chinese herbal formula, which is mainly used in the treatment of concussion, cerebral post-traumatic syndrome. Although its effectiveness has been certified in the clinical use, the mechanism of action and bioactive components of Naozhenning remain unknown. In this study, the ultrahigh performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry was applied to identify the absorbed constituents of Naozhenning in the rat serum. A total of 60 compounds, including 30 prototype components and 30 metabolites were identified. Then the absorbed constituents were subjected to the network pharmacology analysis. The compound-target-disease (CTD) and KEGG pathway analysis revealed that 40 absorbed constituents, 56 target genes and three key pathways such as RAS, PI3K/Akt, MAPK, TGFß were probably related with the efficacy of Naozhenning against cerebral trauma and cerebral concussion. The results provided a scientific basis for understanding the bioactive compounds and the pharmacological mechanism of Naozhenning granule.

Analysis of quality marker in Naozhenning Granule based on “five principles” / 中草药

Naozhenning Granule (NG) is a traditional Chinese medicine prescription, which can cool and activate blood, resolve stasis and dredge collaterals, enrich blood and tranquilize mind, calm nerves and arrest vomiting. The clinical studies have also confirmed that NG was effective for the treatment of headache, dizziness, irritability, insomnia, forgetfulness, nausea, and vomiting caused by cerebral trauma. The quality markers (Q-markers) of NG are predicted and analyzed based on the “five principles” of Q-markers of Chinese materia medica and the research literatures in the recent years. The modern pharmacological research of the selected Q-markers was reviewed and analyzed. Then the network of “Q-marker, targets, pathway, and disease” was constructed, in order to provide the scientific basis for the quality control study of NG.

Identification of chemical constituents in Naozhenning granule by UHPLC coupled with hybrid quadrupole-orbitrap MS / 药学学报

Naozhenning granule, which is composed of 11 herbal drugs, is mainly used in the treatment of concussion, cerebral post-traumatic syndrome. As the chemical composition of Naozhenning was complex, the ultrahigh performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry was applied to identify the chemical constituents rapidly in the Naozhenning granule. By analyzing the retention behaviors, accurate molecular weight, the fragmentation pathway, and comparison with the MS data from the standards and references, as well as the automatic identification by the Compound Discover 2.0, a total of 161 compounds were identified or tentatively deduced, and the sources of constituents were also confirmed. The compounds identified in Naozhenning granule included 9 iridoids, 8 butylphthalides, 26 flavonoids and flavonoid glycosides, 8 phenolic acids, 8 monoterpenoid glycosides and 9 alkaloids, as well as the common compounds in the herbal drugs, such as organic acids, amino acids, and sugars. The chemical composition of Naozhenning was studied for the first time, which provides a scientific basis for the quality control and document of effective materials of Naozhenning granule.

Network pharmacology based study on mechanism of Naozhenning Granule for treatment of cerebral trauma / 中草药

Objective: To predict the targets of the main ingredients in Naozhenning Granule and explore its molecular mechanism of multi-components, multi-targets, and multi-pathways. Methods: Reverse molecular docking (DRAR-CPI) and CooLGeN database were used to predict and screen the targets of Naozhenning Granule; GO enrichment was performed in ClueGO of Cytoscape; KEGG pathway analysis was conducted in DAVID database; The herbs-ingredients-targets-pathways-disease network was constructed in the Cytoscape software. Results: A total of 33 candidate compounds were screened out, and a total of 34 potential targets were revealed for Naozhenning Granule, such as MAPK1, CASP3, and GSK3B. The results of GO enrichment and KEGG pathway analysis indicated that Naozhenning Granule was involved in a series of biological process, such as reactive oxygen species biosynthetic process and positive regulation of smooth muscle cell proliferation as well as some signaling pathways, including PI3K/Akt, MAPK, JAK/STAT, and mTOR. The herbs-ingredients-targets-pathways-disease network suggested that the mechanism of Naozhenning Granule was involved with the regulation of oxidative stress, inhibiting the inflammatory response and the apoptosis of neural cells, regulation of the formation of H2S and the activity of PLG, improving the cognitive function and post traumatic depression. Conclusion: The study suggested that the molecular mechanism of Naozhenning Granule was related with the multi-components, multi-targets, and multi-pathways, which provided a scientific basis for further elucidation of the active ingredients and pharmacological action of Naozhenning Granule.