[Chemical constituents and anti-liver fibrosis mechanism of Meconopsis quintuplinervia based on UPLC-Q-Exactive-MS/MS and network pharmacology].

In this study, UPLC-Q-Exactive-MS/MS was used to rapidly analyze the chemical constituents of Meconopsis quintupli-nervia, and the anti-liver fibrosis mechanism of M. quintuplinervia was preliminarily analyzed by network pharmacology, molecular docking, and cell experiments. The chemical constituents of M. quintuplinervia were identified according to the information of MS~1 and MS~2, as well as the data in the literature and databases. SwissTargetPrediction and TargetNet were used to predict the potential targets. The targets related to liver fibrosis were collected from GeneCards and OMIM. The protein-protein interaction(PPI) network was constructed by STRING. Cytoscape 3.6.1 was used to construct and analyze the "constituent-target-disease" network to obtain key targets and their corresponding constituents in the network. DAVID 6.8 was used for GO analysis and KEGG signaling pathway enrichment analysis. Finally, the preliminary verification was carried out by molecular docking and cell experiments. As a result, 106 chemical constituents were identified from M. quintuplinervia, including 66 flavonoids, 16 alkaloids, 18 phenolic acids, 1 anthocyanin, and 5 other constituents. Among them, 3 constituents were identified as potential new compounds, and 59 constituents were reported in M. quintuplinervia for the first time. Network pharmacology analysis showed that M. quintuplinervia presumably acted on AKT1, SRC, JUN, EGFR, STAT3, HSP90 AA1, MAPK3, and other core targets through luteolin, isorhamnetin, quercetin, apigenin, kaempferide, amurine, 2-methylflavinantine, allocryptopine, the multi and other active compounds, thereby regulating the PI3 K/AKT signaling pathway, pathways in cancer, proteoglycans in cancer, FoxO signaling pathway, and other pathways to exert anti-liver fibrosis effects. M. quintuplinervia extract(MQE) could significantly down-regulate PI3 K and AKT protein levels in the HSC-T6 cell model induced by TGF-ß1, suggesting that MQE may have the ability to regulate the PI3 K/AKT signaling pathway. The findings of this study indicated that the anti-liver fibrosis effect of M. quintuplinervia had multi-constituent, multi-target, and multi-pathway characteristics, which may provide a scientific basis for the research on the pharmacodynamic materials, action mechanism, and quality markers of M. quintupli-nervia.

[Chemical constituents of phenylethanoid glycosides from Forsythiae Fructus and their antitumor activities].

Four phenylethanoid glycosides were isolated from the 75% EtOH extract of Forsythiae Fructus by various column chromatography methods(MCI, silica gel, ODS and semi-preparative HPLC). Their structures were identified as forsythenside M(1), forsythenside K(2), forsythoside I(3) and forsythoside A(4) by physicochemical properties and extensive spectroscopic analysis(UV, 1 D and 2 D NMR, HR-ESI-MS). Among them, compound 1 was one new phenylethanoid glycoside. The in vitro cytotoxic activities of these compounds against MCF-7, A-375, SGC-7901 and B16 F10 were evaluated. The results showed that compounds 1-4 had cytotoxic activities against MCF-7, A-375, SGC-7901 and B16 F10 at 40 µmol·L~(-1).

Comprehensive chemical profiling of monascus-fermented rice product and screening of lipid-lowering compounds other than monacolins.

ETHNOPHARMACOLOGICAL RELEVANCE: Monascus-fermented rice product (MFRP) has been regarded as a dietary supplement and traditional medicine with circulation-promoting effects in China and other countries for centuries. AIM OF THE STUDY: This study was carried out to profile the chemical components in MFRP, and provide available information for elucidating the potential lipid-lowering compounds other than monacolins. MATERIALS AND METHODS: High-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) and gas chromatography coupled with mass spectrometry (GC-MS) methods were applied to comprehensive analysis of chemical components in MFRP. Potential small molecules were identified by comparing with reference standards, or tentatively characterized by comparing their retention time and high-resolution mass spectral data with previous literature. The lipid-lowering properties of ten major non-monacolin compounds were evaluated in cholesterol-fed zebrafish larvae. And one with optimum lipid-lowering activity was subsequently evaluated in high fat diet-fed C57BL/6 J mice, with the dyslipidemia and ectopic lipid deposition being investigated. RESULTS: A total of 99 compounds were characterized in MFRP, including 38 monacolins, 5 decalins, 6 isoflavones, 13 pigments, 8 azaphilonoids, 11 amino acids, 4 nucleosides, 9 lipid acids, 4 phytosterols and glycerol. The preliminary screening showed that ergosterol remarkably reduced cholesterol levels in zebrafish larvae. Moreover, ergosterol delayed body weight gain and decreased circulating total cholesterol, triglyceride, low density lipoprotein cholesterol levels in high fat diet-fed mice. Ectopic lipid accumulation was also ameliorated in the liver and heart of obese mice. CONCLUSION: Global analysis of chemical components and screening of lipid-lowering non-monacolin compounds in MFRP have improved our understanding of its therapeutic material basis.

[Determination of quercetin and kaempferol in Fagopyrum spp. of different species and different parts by HPLC].

OBJECTIVE: An HPLC method was established for determination of quercetin and kaempferol in Fagopyrum spp. of differrent species and different plant parts. METHODS: The chromatographic separation was carried out on a DIKMA diamonsil (250 mm x 4.6 mm, 5 µm) chromatographic column. The mobile phase was acetonitrile (A) and 0.1% phosphoric acid solution (B) at the flow rate of 1.0 mL/min. The detection wavelength was set at 260 nm and the column temperature was 25 degrees C. RESULTS: The descending order of quercetin content in the samples of different parts of Fagopyrum tataricum was seed (6.12 mg/g) > leaf (1.47 mg/g) > stem (0.34 mg/g); quercetin was not found in root. Only a few kaempferol was found in seed (0.42 mg/g) and leaf of Xiqiao 1 (0.09 mg/g) of Fagopyrum tataricum. There were a few quercetin was determined in the whole plant of Fagopyrum esculentum, and kaempferol was not been found. The total and mean value concentration of quercetin and kaempferol of Fagopyrum tataricum were much higher than that of Fagopyrum esculentum. In different species of Fagopyrum tataricum planted in the same area, the content of quercetin and kaempferol was similar, they were not been found in Fagopyrum esculentum. CONCLUSION: This method is simple, rapid, accurate and reliable for quality evaluation of Fagopyrum spp. and its commercial products.

[Influence of different processed methods on 10 kinds of ginsensides in Panacis Quinquefolii Radix].

The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.