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Outline
In this study, the formation mechanism of the nanophase of "Rhizoma Anemarrhenae-Gypsum" was investigated, and its compatibility, antipyretic efficacy, and underlying mechanisms were systematically evaluated via network pharmacology. Three groups, namely the Rhizoma Anemarrhenae group, the Gypsum group, and the Rhizoma Anemarrhenae-Gypsum group, were set, with all groups including both nanophase preparations and traditional decoctions. The particle size, salinity, electrical conductivity and surface tension of the solutions were determined, and these combined with transmission electron microscope (TEM) and fingerprint analysis. Targets were screened using the GeneCards and DisGeNet, an active component-target network was constructed, a PPI network was built with the help of the STRING database and Cytoscape, and GO and KEGG enrichment analysis was performed using Metascape. The antipyretic effect of the nanophase solution was verified by hematoxylin-eosin (HE) staining, Western blot and real-time PCR. The results indicated that salinity and electrical conductivity promoted the stable formation of the nanophase; and TEM revealed that timosaponin coated the Gypsum crystal structure, leading to nanoparticle formation. Fingerprint analysis identified eight common peaks (neomangiferin and mangiferin as the main components), and the chemical components of the nanophase were consistent with those of the decoction. Through network pharmacology, 31 active components of Rhizoma Anemarrhenae and Gypsum were obtained, corresponding to 138 potential targets; the number of fever-related targets was of 1 093 fever-related targets were retrieved, among which 188 overlapping targets were screened. The main KEGG enrichment pathways included epidermal growth factor receptor, interleukin-17, hypoxia-inducible factor 1 and other signaling pathways. Experimental results showed that the nanophase group of Rhizoma Anemarrhenae-Gypsum decoction significantly reduced the body temperature in febrile rats, inhibited the expression of inflammatory factors, down-regulated fever medium, up-regulated cytoprotective proteins, and suppressed the expression of related mRNA. These findings were consistent with the predictions of network pharmacology. All animal experiments in this study were approved by the Animal Experimental Ethics Committee of Harbin University of Commerce (HSDYXY-2024022). This study found that Rhizoma Anemarrhenae-Gypsum enhances its therapeutic efficacy through the nanophase, and its antipyretic mechanism involves the multi-target regulation of inflammatory pathways and central signaling molecules thereby providing both theoretical and experimental support for the modernization of traditional Chinese medicine compound prescriptions.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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