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Outline
Based on network pharmacology and molecular docking technology， the key targets of the main active components of Portulaca oleracea L. （POL） to inhibit mastitis in dairy cows were explored， and the possible mechanism of action was verified by in vitro cell experiments. The TCMSP database was used to search the active ingredients and corresponding targets of POL. The GeneCards database and OMIM database were used to search the disease targets of dairy cow mastitis， and the intersection and integration of the active ingredient targets of POL were carried out， and the PPI network was constructed. DAVID database was used for GO function enrichment analysis and KEGG pathway enrichment analysis， and AutoDock software was used for molecular docking verification. Finally， the effect of purslane extract on bovine mammary epithelial cells was determined by CCK-8 method. LPS induced MAC-T to construct an in vitro inflammation model. RT-qPCR was used to detect the expression of key targets， IL-17 signaling pathway and inflammation-related factors. Results showed that a total of 10 active components and 188 corresponding targets of POL were screened. A total of 2 495 targets related to mastitis in dairy cows were screened from the GeneCards and OMIM databases， and a total of 40 co-expressed targets were found through the Venn platform. Ten key targets such as ALB， TNF， IL6， IL1 B， and MMP9 were obtained through the PPI network， involving AGE-RAGE， lipid and atherosclerosis， cancer， IL-17， PI3K-AKT and other signaling pathways. In vitro cell experiments showed that purslane could increase the content of ALB in MAC-T， and provide IL-17 signaling pathway to down-regulate the transcription of inflammatory factors MMP9， TNF-α， IL-6， IL-1β， IL-8 and IFN-γ genes， and up-regulate the expression of anti-inflammatory factor IL-10 gene. This study revealed that purslane can inhibit the release of inflammatory factors by inhibiting the IL-17 signaling pathway， thereby inhibiting the inflammatory response of dairy cow mammary gland.
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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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