Discovery and mechanistic exploration of anti-colorectal cancer bioactive compounds from Perilla frutescens leaf volatile oil by integrating GC-MS, network pharmacology, and pathway validation

The aim of this study is to analyze the chemical composition of the volatile oil from Perilla frutescens leaves and explore the pharmacological substances and mechanisms of action of Perilla frutescens leaf volatile oil in combating colorectal cancer (CRC). Using dried leaves of Perilla frutescens as raw material, the volatile oil was extracted by steam distillation, and the chemical composition of the volatile oil was identified by gas chromatography-mass spectrometry (GC-MS). Exploring the potential target proteins and mechanisms of action of perilla leaf volatile oil using network pharmacology and molecular docking. The cell counting kit-8 (CCK-8) assay was used in vitro to determine the inhibitory effect of volatile oil active ingredients on the proliferation of human colon cancer HCT116 cells; Real time fluorescence quantitative PCR and protein immunoblotting experiments were used to detect the expression of mRNA and pathway related proteins of potential targets. Thirty chemical components were identified in the volatile oil of Perilla frutescens leaves. The results of network pharmacology showed that the volatile oil from Perilla frutescens leaves regulates phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) and mitogen activated protein kinase (mitogen activated protein kinase) by acting on targets such as heat shock protein 90 alpha family class A member 1 (HSP90AA1), proto oncogene tyrosine kinase src (SRC), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), etc. The protein kinase (MAPK) signaling pathway plays an anti CRC role. Molecular docking revealed that representative components (perilla aldehyde, celery brain, trans orange blossom tertiary alcohol) have strong affinity for potential anti CRC targets. The cell experiment results showed that trans orange blossom tertiary alcohol and celery brain had significant inhibitory effects on the proliferation of HCT116 cells. Perilla leaf volatile oil inhibited EGFR mRNA expression (P