Andrographolide and Berbamine Synergy in Glioblastoma Treatment: An Insight into the Pathways Assimilating Proteomics and Metabolomics

Glioblastoma (GB) is one of the most aggressive and invasive cancers, characterized by poor survival rates and high recurrence. Standard treatments concerns about toxicity and long-term safety have led to growing interest in natural alternatives, such as phytotherapeutics. The study explores the effects of two phytocompounds: andrographolide, a diterpenoid, and berbamine, an alkaloid, on GB cells. Their cytotoxic effects were tested on GB cell lines LN229 and U-87 MG and HEK 293 cells to assess safety. Additionally, the impact of these compounds on wound healing, apoptosis (via Annexin V), cell cycle progression, oxidative stress, and mitochondrial dysfunction was evaluated in GB cells. The DSC, mass spectrometry and NMR along with ontology analysis were used to study the changes in protein structure and levels leading to changes in metabolism. The endothermic and exothermic events, highlighted shifts in thermal stability between treated and control cell lines. While, several proteins showed significant change in levels in response to the combination treatment of chosen phytocompound combination. Finally, the treatment with the combination caused significant changes in the metabolic profile of glioblastoma cells. The results highlight the strong potential of phytocompounds combination to target GB by targeting multiple pathways for GB.Statement of significance of the studyGlioblastoma (GB) is one of the deadliest brain cancers. It has few treatment options and a poor prognosis because of tumor recurrence, resistance to therapy, and the toxicity of standard treatments. This study offers new insights into the potential of two plant compounds, andrographolide and berbamine, as a combination treatment for GB. Our results show that the combination produces stronger effects, increases cell death, raises mitochondrial oxidative stress, and causes cell cycle arrest, which reduces GB cell growth and movement. In addition, analysis through computer simulation revealed disruptions in tumor-related signaling pathways and metabolic processes. Proteomic and metabolomic profiling showed notable changes in energy metabolism, amino acid turnover, and lipid production. These findings suggest that the combination of these plant compounds targets several key features of glioblastoma. Moreover, it could disrupt the metabolism enough to overcome resistance to standard therapies. Importantly, safety tests on HEK293 cells indicate that the treatment selectively harms GB cells. Overall, this research highlights the importance of using natural compounds in multi-target treatment strategies. It points to andrographolide and berbamine as promising candidates for treating glioblastoma, which calls for more investigation.