Comprehensive network pharmacology and experimental study to investigate the effects and mechanisms of Lophatherum gracile Brongn. for glioma treatment.

Background: Gliomas are challenging to treat due to their invasive nature and resistance to conventional therapies. Recent study has revealed that Lophatherum gracile Brongn. extract has anti-cancer properties on fibrosarcoma cell. Nevertheless, its medicinal effects and mechanistic pathways on gliomas have not been explored.

Aim Of The Study: To investigate the detailed anti-glioma roles of Lophatherum gracile Brongn. extract and its specific pharmacological routes of action both in vitro and in vivo, focusing on autophagy, apoptosis, proliferation, and migration.

Methods: Ethanol extracts of Lophatherum gracile Brongn. were prepared, and the active compounds were appraised by high-performance liquid chromatography. Human glioma cells (A172, LN229, U-87 MG, and U251) were treated with various concentrations of the extract. Immunofluorescence and transmission electron microscopy were employed to investigate the autophagosomes. Cell viability, proliferation, migration, and death were assessed using various assays. The anti-tumor effects were further tested in animal models. Network pharmacology was employed to investigate the potential targets of the main compounds acting on glioma. Furthermore, RNA sequencing was conducted for transcriptome analysis and molecular docking was used for identification.

Results: The extract significantly attenuated glioma cell viability, proliferation, and migration, both in vitro and in vivo, while promoting autophagy, apoptosis, and cell death. Five active compounds were identified: chlorogenic acid, isoorientin, orientin, vitexin, and isovitexin. A total of 1472 glioma targets were identified, along with 219 drug targets for the five compounds. Combining analysis of differentially expressed genes revealed that dihydrofolate reductase may be a key target of these compounds, as determined by molecular docking analysis.

Conclusions: Lophatherum gracile Brongn. extract exhibits significant anti-glioma effects in both cellular and animal models by targeting dihydrofolate reductase, providing novel insights into its therapeutic potential for glioma.