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To predict potential quality markers(Q-Marker) of Taxus media Rehd. based on fingerprint and network pharmacology methods, and establish an evaluation method of Taxus media Rehd. based on Q-Marker, as so to provide basis for the establishment of quality standard and quality evaluation system of Taxus media Rehd..
Waters SymmmetryShieldTM RP18(250 mm×4.6 mm, 5 μm) chromatographic column was used for separation, and the wavelength was 254 nm. The mobile phase was acetonitrile -0.01% trifluoroacetic acid aqueous solution for gradient elution at a flow rate of 1.0 mL·min-1, and the column temperature was 30 ℃. The injection volume was 10 μL. The fingerprints of 24 batches of Taxus media Rehd. with 8 kinds of cultivation years were established and the common peaks were identified. Hierarchical clustering analysis (HCA) was used for classification, orthogonal partial least square-discriminant analysis (OPLS-DA) was used to screen out the main marker components that cause differences between different years. Combined with network pharmacology, the core targets and key pathways were constructed a “component-target-pathway” network map through corresponding databases. Q-Markers were further verified and quality of Taxus media Rehd. comprehensively evaluated by HCA and PCA.
A total of 25 common peaks were obtained in the HPLC fingerprint of Taxus media Rehd., 18 compounds were identified, and their similarities were all above 0.900, including taxanes, flavonoids, alkaloids, steroids and phenols. The results of HCA showed that there were some differences among samples with different cultivation years. Eleven different markers were screened by OPLS-DA, and they were paclitaxel, 10-deacetylbaccatin Ⅲ (10-DAB), baccatin Ⅲ, cephalomannine, deacetyltaxol (10-DAT), ferulic acid, kaempferide, rutin, amentoflavone-4’, 4", 7-trimethyl ether, skyrin, 4-hydroxybenzaldehyde. Based on the difference markers, network pharmacology was applied to analyze them from the perspective of effectiveness. Paclitaxel, 10-DAB, bakatine Ⅲ, cephalomannine and 10-DAT were preliminstly predicted as Q-Markers of Taxus media Rehd.. They could regulate cancer pathway, AGE-RAGE signaling pathway in diabetic complications and other signaling pathways by PIK3R1, AKT1, EGFR, HRAS and MAPK1 targets, and play the role of reducing swelling and menstruation diuresis clearing. The results of HCA verified the rationality of Q-Marker, and the comprehensive evaluation results of PCA showed that sample which was cultivated for ten years and harvested in spring bore the best quality.
Paclitaxel, 10-DAB, bakatine Ⅲ, cephalomannine and 10-DAT are selected as Q-Markers based on fingerprint and network pharmacological methods. The quality of Taxus media Rehd. in different cultivation years is sorted comprehensively by combining with chemical recognition pattern method. The 10-year-old sample harvested in spring gets the highest score and the quality is good. The study provides new method for establishing of quality standard and the quality evaluation of Taxus media Rehd..
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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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