Spatially resolved metabolomics combined with bioactivity analyses to evaluate the pharmacological properties of two Radix Puerariae species

Ethnopharmacological relevance: P. lobata and P. thomsonii are medicinal plants with similar pharmacological functions but different therapeutic effects. A novel method is presented herein to investigate metabolites in terms of their distribution and qualification, quantification is necessary to elucidate the different therapeutic effects of the two Puerariae species. Aim of the study: The aim of the present study was to perform spatially resolved metabolomics combined with bioactivity analyses to systematically compare the metabolite differences in P. lobata and P. thomsonii by dis-tribution, qualification, quantification, and biological activity to evaluate their pharmacological properties. Materials and methods: Air flow-assisted desorption electrospray ionization-mass spectrometry imaging (AFA-DESI-MSI) was performed to characterize the differences in the metabolite distributions of P. lobata and P. thomsonii. Further qualitative and quantitative analyses of the differential metabolites were performed using liquid chromatography-mass spectrometry (LC-MS). Biological activities correlated with the differences in the metabolites were validated by MTT assays. Results: Some metabolites showed complementary distributions of the phloem and xylem in the two species, saccharide, vitamin, and inosine levels were higher in the phloem of P. thomsonii but higher in the xylem of P. lobata. The 3 '-hydroxyl puerarin level was higher in the xylem of P. thomsonii but higher in the phloem of P. lobata. Qualitative and quantitative analyses of the metabolites revealed a total of 52 key differential me-tabolites. MTT assays showed that daidzein, daidzin, puerarin, ononin, genistin, formononetin, 3 '-hydroxy puerarin, 3 '-methoxy puerarin, mirificin, and 3 '-methoxy daidzin exerted protective effects on H9c2 cells against hypoxia/reoxygenation injury. P. lobata extracts exhibited a significantly better protective efficacy than P. thomsonii extracts. Conclusions: In this study, AFADESI-MSI combined with LC-MS and biological activities comprehensively eluci-dated metabolite differences in the distribution, qualification, quantification, and pharmacological properties of P. lobata and P. thomsonii. The results of this study could provide a novel strategy for species identification and quality assessment of similar Chinese herbal medicines.

Automated summary

This study systematically compared the metabolite differences between P. lobata and P. thomsonii using AFA-DESI-MSI, LC-MS, and biological activity analysis. The results revealed 52 key differential metabolites and showed that P. lobata extracts exhibited a better protective efficacy on H9c2 cells against hypoxia/reoxygenation injury compared to P. thomsonii extracts.