Optimization of Extraction Method of Ginkgolide and Its Regulation of Oxidative Stress in Liver Cancer

Ginkgolide （GG） is mainly composed of sesquiterpene lactones and diterpene lactones, which have great potential for development and area hot research topic in pharmacology. However, the extraction process of natural active components of ginkgo esters needs to be further explored, and there are limited reports on the pharmacological activities of its main components, ginkgolides A/B/C and bilobalides in the treatment of liver cancer. Supercritical extraction combined with chromatographic separation were utilized for the purification of Ginkgo biloba leaf/G. biloba extract to investigate its pharmacological activity. Modulation of oxidative stress was explored in liver cancer through network pharmacology analysis and vitro cell experiments, and further an analytical chemistry pharmacology cell biology correlation model was established to investigate the pharmacological characteristics of ginkgolide. The results showed that the optimal conditions for supercritical extraction and separation were 37.6 ℃, pressure 35 MPa, CO₂ flow rate 8 L·h⁻¹, and extraction time 150 min, in the circumstances ginkgolide A/B/C and bilobalide were obtained with their purity of 98.0% after chromatographic purification. Network pharmacology analysis showed that the four extracts could regulate liver cancer through core targets such as IL2, MAPK14, GSK3B, STAT1 and SRC that mainly involved the MAPK signaling pathway, Rap1 signaling pathway, viral carcinogenesis, hepatitis C and alcoholic liver disease, all of which were related to the development of cancer, cell apoptosis and liver disease. Ginkgolide acted on SMMC-7721 liver cancer cells, as the dose increased, cell survival rate decreased, SOD and CAT enzyme activities also decreased, and LDH enzyme activity increased. This indicated that ginkgolide could cause insufficient clearance of H₂O₂ and O₂⁻ by inhibiting the activities of SOD and CAT enzymes, and ROS attacked membrane lipids, leading to membrane rupture and LDH leakage, ultimately resulting in cell apoptosis. This result not only optimizes the efficient preparation process of active ingredients in G. biloba, but also explores the effects from the perspectives of drug network target analysis and cellular oxidative stress, providing reference for the deep development of G. biloba resources and new strategies for liver cancer treatment.