Network pharmacology to investigate the pharmacological mechanisms of muscone in Xingnaojing injections for the treatment of severe traumatic brain injury

Zhuohang Liu; Hang Li; Wenchao Ma; Shuyi Pan

doi:10.7717/peerj.11696

Network pharmacology to investigate the pharmacological mechanisms of muscone in Xingnaojing injections for the treatment of severe traumatic brain injury

PeerJ. 2021 Jul 20:9:e11696. doi: 10.7717/peerj.11696. eCollection 2021.

Authors

Zhuohang Liu^#^{1

2}, Hang Li^#², Wenchao Ma^{1

3}, Shuyi Pan^{1

2}

Affiliations

¹ The Fifth Clinical Medical College of Anhui Medical University, Beijing, China.
² Department of Hyperbaric Oxygen, Sixth Medical Center, Chinese PLA General Hospital, Beijing, China.
³ Department of Neurology, Sixth Medical Center, Chinese PLA General Hospital, Beijing, China.

^# Contributed equally.

Abstract

Background: Xingnaojing injections (XNJI) are widely used in Chinese medicine to mitigate brain injuries. An increasing number of studies have shown that XNJI may improve neurological function. However, XNJI's active ingredients and molecular mechanisms when treating traumatic brain injury (TBI) are unknown.

Methods: XNJI's chemical composition was acquisited from literature and the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. We used the "absorption, distribution, metabolism, and excretion" (ADME) parameter-based virtual algorithm to further identify the bioactive components. We then screened data and obtained target information regarding TBI and treatment compounds from public databases. Using a Venn diagram, we intersected the information to determine the hub targets. Cytoscape was used to construct and visualize the network. In accordance with the hub proteins, we then created a protein-protein interaction (PPI) network using STRING 11.0. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were analyzed according to the DAVID bioinformatics resource database (ver. 6.8). We validated the predicted compound's efficacy using the experimental rat chronic constriction injury (CCI) model. The neuronal apoptosis was located using the TUNEL assay and the related pathways' hub proteins were determined by PCR, Western blot, and immunohistochemical staining.

Results: We identified 173 targets and 35 potential compounds belonging to XNJI. STRING analysis was used to illustrate the protein-protein interactions and show that muscone played a fundamental role in XNJI's efficacy. Enrichment analysis revealed critical signaling pathways in these components' potential protein targets, including PI3K/AKT1, NF-kB, and p53. Moreover, the hub proteins CASP3, BCL2L1, and CASP8 were also involved in apoptosis and were associated with PI3K/AKT, NF-kB, and p53 signaling pathways. We showed that muscone and XNJI were similarly effective 168 h after CCI, demonstrating that the muscone in XNJI significantly attenuated neuronal apoptosis through the PI3K/Akt1/NF-kB/P53 pathway.

Conclusion: We verified the neuroprotective mechanism in muscone for the first time in TBI. Network pharmacology offers a new approach for identifying the potential active ingredients in XNJI.

Keywords: Muscone; Network analysis; Protein targets; Traumatic brain injury; XNJI.

Grants and funding

This research received support from the Research Fund of Chinese PLA General Hospital-Sixth Medical Center (CXPY201816), the Beijing Nova Program (Z171100001117092), the National Nature Science Foundation of China (81500669) and the National Nature Science Foundation of Beijing (7154241). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.