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Objective:To evaluate the anti-inflammatory potential of aqueous extract of Pterocarpus santalinus L.f. heartwood using molecular docking and in vivo experiment. Methods:An aqueous extract of Pterocarpus santalinus heartwood was prepared using a Soxhlet apparatus. Phytocompounds in the extract were tentatively identified using high-resolution mass spectrometry. Molecular docking experiments were carried out to evaluate the binding affinity of selected compounds, phloridzin to cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), prostaglandin E synthase-1 (PGES-1) and 5-lipoxygenase (5-LOX). Anti-inflammatory potential was evaluated by carageenan induced paw edema model in rats. Results:The presence of major component phloridzin along with quercetin, parthenin, ginkgolide B, picrotoxinin, usnic acid, octopine, and epigallocatechin was detected in the extract. Molecular docking study showed that phloridzin inhibited COX-1, COX-2, PGES-1 and 5-LOX with more affinity than ibuprofen and paracetamol. Pterocarpus santalinus heartwood extract at 200 and 400 mg/kg BW showed significant reduction in carageenan-induced hind paw edema in a dose-dependent manner, but the effect was slow when compared with the standard ibuprofen (30 mg/kg p.o.). Conclusions:The study indicated that after clinical trials, the aqueous extract of Pterocarpus santalinus heartwood can be effectively used in phytotherapy to treat inflammation.
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Objective: To clone the leucoanthocyanidin reductase (LAR) gene of Lithocarpus polystachyus, and analyze the relationship between LAR gene expression level and phloridzin content. Methods: According to the results of L. polystachyus transcriptome sequencing (unigene: DN30711_c0_g1_i1), the full-length cDNA sequence of LAR gene was amplified by PCR and the bioinformatics analysis was carried out. Its expression was detected by quantitative Real-time PCR (qRT-PCR). The phloridzin content of L. polystachyus was measured by UPLC method and the correlation between LAR gene expression and phloridzin content was analyzed by SPSS 18.0 software. Results: The full-length cDNA of the LAR gene was 1 053 bp and contained a complete open reading frame that encoded 350 amino acids. This protein did not exist a transmembrane domain and was localized in the cytoplasm. The LAR protein of L. polystachyus was the number of PCBER_SDR_a family and had a high similarity (95%) to the LAR protein of Quercus suber and their genetic relationship was close. The phloridzin content of L. polystachyus was positively correlated with the expression of LAR gene (P < 0.05). Conclusion: The LAR gene of L. polystachyus was cloned for the first time. It was confirmed that the content of phloridzin was positively correlated with the expression of LAR gene of L. polystachyus, which laid a theoretical and technical basis for revealing the biosynthesis mechanism of phloridzin of L. polystachyus.
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Objective: To screen the flavonoid constituents and targets of Litchi Semen in the intervention of progression and metastasis of colon adenocarcinoma (COAD). Methods: Through DRAR-CPI and SWISS database, potential targets of 19 flavonoids in Litchi Semen were searched. COAD gene expression data and clinical characteristic data from TCGA database were downloaded. Weighted gene co-expression network analysis (WGCNA) was used to establish the gene co-expression network and identify the co-expression module of COAD. The common targets of co-expression module and potential targets were used as the compound to interfere with the drug target of COAD. Protein interaction network analysis, KEGG and GO analysis were performed by String database. The Hub gene was extracted as potential biomarkers of COAD by the cytoHubba, and the interaction network of components, targets and pathways was established by the Cytoscape. The expressions of potential biomarkers were verified by HPA database, and the compounds were docked with the potential biomarkers. Results: A total of 18 co-expression modules were identified with seven of them were correlated with clinical features, such as survival time and tumor stage. Turquoise module was related to the development and transfer of COAD. 19 flavonoids in Litchi Semen acted on 380 potential targets. 34 targets repeated with turquoise module were selected as targets. GO analysis showed that the target points were enriched in 304 GO items, including 229 biological processes, 31 cell composition and 44 molecular functions; KEGG analysis showed that target points were enriched in cancer pathways, cell cycle, and progesterone-mediated 40 pathways including oocyte cancer pathway, cell senescence, and p53 signaling pathway. The genes of CDC25A, CDC25C, CCNB2 and AURKB were screened by cytoHubba as potential biomarkers which related to the progress and transfer of COAD. Compared with para-cancerous tissues, immunohistochemistry results obtained from HPA database showed that the protein expressions of CDC25C, AURKB and CCNB2 in COAD were increased significantly (P < 0.05), which were consistent with gene expression in TCGA data set. Narirutin, procyanidin A2, phloridzin and ent-epicatechin which were well combined to CDC25A, CDC25C and AURKB through hydrogen bond were screened. Conclusion CDC25A, CDC25C, CCNB2 and AURKB were the potential biomarkers closely related to the progression and metastasis of COAD. The mechanism of intervention of flavonoids in Litchi Semen on the progression and metastasis of COAD may be related to the regulation of biological processes, such as cell division, G2/M phase transformation of cell cycle, and the regulation of cancer pathway, p53 signaling pathway and other signaling pathways. Narirutin, procyanidin A2, phloridzin, ent-epicatechin and rutin could be treated as potential inhibitors of CDC25A, CDC25C and AURKB.
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To establish a method for the simultaneous determination of phloridzin, 3-hydroxy phloridzin and quercitrin in leaves of Malus halliana by ultrasonic-assisted ionic liquid coupled with RP-HPLC. An Agilent TC-C₁₈ (4.6 mm×250 mm, 5 μm) column was used, with the mobile phase of acetonitrile and 1% phosphoric acid-water (20∶80) by gradient elution at the detection wavelength of 270 nm. The flow rate was 0.8 mL•min⁻¹, and chromatographic column temperature was controlled at the room temperature. Under the optimized conditions, the linear ranges for phloridzin, 3-hydroxy phloridzin and quercitrin were 0.9-112.5 μg (r = 0.999 6), 0.093 2-11.65 μg (r = 0.999 1) and 0.097 2-12.15 μg (r = 0.999 8), respectively. The average recoveries of the three constituents were 99.35%, 98.80% and 98.19%, respectively. The method was environmental friendly, rapid, accurate and highly reproducible, and so suitable for the quantitative analysis of phloridzin, 3-hydroxy phloridzin and quercitrin in leaves of M. halliana.
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To study the in vivo intestinal absorption kinetics of phloridzin in rats. The absorption of phloridzin in the small intestines and colon of rats was investigated using an in vivo single-pass perfusion method and the drug concentration was measured by HPLC. The effects on intestinal absorption of different drug concentration and P-glycoprotein (P-gp) inhibitor were conducted. The results showed that the phloridzin could be absorbed in whole intestine, but more fully in the jejunum and colon segment,poorly absorbed in the duodenum and ileum. The absorption rate constant (Ka) and the apparent absorption coefficient（Papp）of phloridzin decreased following the sequence of jejunum> colon > duodenum > ileum. Absorption parameters of phloridzin had no significant difference at different concentration (5.14, 10.28, 20.56 mg•L⁻¹) . The saturate phenomena was not observed under the test range of drug concentration, and the absorption mechanism may be the passive diffusion transport.There had a significant difference in Ka and Papp values between P-gp inhibitor and no P-gp inhibitor groups. Phloridzin may be the substrate of P-gp.
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Objective To establish a reversed-phase high performance liquid chromatograph ( RP-HPLC ) method for determination of equilibrium solubility and oil/water partition coeficient of phloridzin in different solvents. Methods A RP-HPLC method was established to detect the concentration of phloridzin in water and different organic solvents. The partition coefficients in the n-octanol-water/buffer solution systems of phloridzin were determined by shaking flask method. The Inertsil ODS-3 (4.6 mm×150 mm, 5μm) column was used and the detection wavelength was 284 nm.The flow rate was 1.0 mL??min-1, and acetonitrile-0.05% phosphoric acid(30??70)was used as mobile phase. Results The equilibrium solubility of phloridzin was 2.07 mg??mL-1 in water and 838.63 mg??mL-1 in methanol at 25 ℃.A good linear relationship of phloridzin was obtained within the range of 0.054 9-1.098 0 μg.The regression equation was Y=2 152.9X+7.26 (r=0.999 9).The solubility values of phloridzin were higher in ethanol and propylene glycol than in other solvents. Conclusion RP-HPLC method is simple, quick and accurate for the determination of phloridzin.Phloridzin was almost insoluble in petroleum ether and poorly soluble in water.The equilibrium solubility is higher in methanol than in other solvents. The apparent distribution coefficient of phloridzin varies significantly with pH under the alkaline conditions but less in the acidic solution.
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Objective: To isolate and identify the phenolic constituents with anti-oxidant and anti-α-glucosidase activities from the methanol extract in the twigs of Acer rubrum. Methods: The twigs of A. rubrum were extracted by methanol then partitioned by system solvents with different polarity. The ethyl acetate extract was separated on silica gel, Sephadex LH-20, ODS columns, and by semi-preparative HPLC. The isolated compounds were identified by physicochemical properties and spectral analyses. The DPPH free radical scavenging and anti-α-glucosidase activities of the compounds were also evaluated. Results: Ten phenolic compounds were isolated and purified from the twigs of A. rubrum and were identified as catechin (1), epicatechin (2), epicatechin-3-O-gallate (3), quercetin-3-O-α-L-rhamnoside (4), quercetin-3-O-3″-galloyl-rhamnoside (5), quercetin-3-O-2″-galloyl-rhamnoside (6), phloridzin (7), ginnalin A (8), ginnalin B (9), and ginnalin C (10). Conclusion: Compounds 3 and 5-7 are isolated from the twigs of A. rubrum for the first time. Compound 7 is the only one chalcone isolated from the plants in Aceraceae for the first time. All of the compounds show the good anti-oxidant activities. Compounds 3 and 8 show the strong anti-α-glucosidase activities.