[Antidepressant active substance basis of ethanol extract from Nardostachyos Radix et Rhizoma based on spectrum-effect relationships and blood-entering component verification].

The antidepressant active substance basis of the ethanol extract from Nardostachyos Radix et Rhizoma was analyzed and interpreted by establishing the high-performance liquid chromatography(HPLC) feature spectra and the spectrum-effect relationship of cell damage protection, and combining the analysis and screening of blood-entering components. HPLC was used to establish feature spectra for 18 batches of Nardostachyos Radix et Rhizoma from different origins. Principal component analysis(PCA) and cluster analysis were used to evaluate and analyze the 18 batches of Nardostachyos Radix et Rhizoma. The cytoprotective effect of 95% ethanol extract of Nardostachyos Radix et Rhizoma was evaluated by using a corticosterone-induced PC12 cell damage model. Gray correlation analysis(GCA), partial least squares regression(PLSR) analysis, and Pearson correlation analysis were used to screen for key active components in Nardostachyos Radix et Rhizoma that protect PC12 cells from damage. Blood-entering components were identified by ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry(UHPLC-MS/MS). The results showed that the similarity of the feature spectra of the 18 batches of Nardostachyos Radix et Rhizoma samples established was all > 0.8, with 36 common peaks calibrated and 12 chemical components identified. The results of cluster analysis and PCA were consistent. The 18 batches of Nardostachyos Radix et Rhizoma samples were divided into three categories. A total of 43 prototype components and one metabolite, mainly flavonoids, terpenoids, phenylpropanoids, alkaloids, fatty acids, and other compounds, were detected in the blood-entering components. The results of the gray correlation analysis showed that the correlation of the 36 common peaks in the Nardostachyos Radix et Rhizoma feature spectrum was all > 0.8. PLSR analysis and Pearson correlation analysis preliminarily identified components such as cryptotanshinone, paeonol, aristolone, nardosinone, and luteolin as the key active substances responsible for improving PC12 cell damage in the ethanol extract of Nardostachyos Radix et Rhizoma. The active components identified through spectrum-effect relationship analysis, including cryptotanshinone, paeonol, nardosinone, luteolin, alpha-asarone, and alpha-cyperone, were all verified in the identification of blood-entering prototype components. The study of the spectrum-effect relationship analysis clarified that the improvement of PC12 cell damage by the 95% ethanol extract of Nardostachyos Radix et Rhizoma was the result of the synergistic action of multiple components. Furthermore, through the verification of blood-entering components, the 11 compounds screened out can serve as the active ingredient group responsible for the antidepressant activity of Nardostachyos Radix et Rhizoma. This provides a research foundation and direction for elucidating the substance basis and mechanism of action of the antidepressant activity of Nardostachyos Radix et Rhizoma, as well as scientific evidence for improving the quality standards related to the "quality-effect" relationship of Nardostachyos Radix et Rhizoma.