Screening the anti-hypoxic active components of Zanthoxylum bungeanum Maxim. seeds based on spectrum-effect...

Zanthoxylum bungeanum Maxim. seeds (ZBS), as an important edible and medicinal resource, possess the efficacy of promoting diuresis, alleviating edema, and relieving dyspnea. It has been reported that these seeds contain unsaturated fatty acids, proteins, and alkaloids, and exhibit diverse pharmacological activities, including antiinflammatory, antioxidant and antibacterial activities.

his study aimed to screen for the components responsible for the anti-hypoxic activity of ZBS using spectrum-effect relationship analysis.

GC-MS fingerprints of ZBS from 24 different origins were first established, and the common peaks were also identified. Anti-hypoxic activity was assessed using a cell injury model, in which PC12 cells were treated with 600 μM CoCl₂·6H₂O to establish a hypoxic condition. Then, the spectrum-effect relationship (SER) between the ZBS fingerprints and anti-hypoxic activity was investigated using Pearson correlation analysis, grey relation analysis (GRA), and orthogonal partial least squares (OPLS) analysis. Finally, potential anti-hypoxic components were screened via SER analysis and further validated using an in vitro assay.

In the GC-MS fingerprint, 18 common peaks were identified from ZBS extracts collected from 24 different origins. Similarity evaluation was performed using hierarchical cluster analysis (HCA) and principal component analysis (PCA), with consistent classification results, the P7 (palmitic acid) and P13 (oleic acid) exhibited higher abundances among the common peaks. Cell experiments indicated that cell viability significantly improved after treatment with different concentrations of ZBS extract. Following the spectrum-effect relationship analysis, GRA results showed that P7, P13, and P14 had the highest correlation with anti-hypoxic activity, with correlation coefficients of 0.964, 0.974, and 0.965, respectively; Pearson correlation analysis revealed that P1, P7, P13 and P14 were significantly positively correlated with anti-hypoxic activity; OPLS results, based on variable importance in projection (VIP) values > 1, demonstrated that P6, P7, P12, and P13 were the key variables in anti-hypoxic activity. Integrating the results from the three analytical approaches, P7 and P13 exhibited a strong contribution to the anti-hypoxic activity of the seeds. Furthermore, the anti-hypoxic activity of both compounds was confirmed through in vitro assays and morphological observations.

This study established the SER between the liposoluble components of ZBS and its anti-hypoxic activity using GC-MS fingerprinting combined with multivariate statistical methods. The key anti-hypoxic constituents were identified as oleic acid and palmitic acid. These findings provide a scientific basis for the development of ZBS-based anti-hypoxic functional foods and pharmaceuticals.