RESUMO
The water extraction and ethanol precipitation method is an extraction method based on the solubility characteristics of polysaccharides that offers wide applicability in the extraction and separation of plant polysaccharides. However, this method leads to large amounts of proteins, nucleic acids, pigments, and other impurities in the polysaccharides products, which makes downstream purification complicated and time-consuming. In this study, a green, high-density natural deep eutectic solvents was used for the high-purity extraction and separation of polysaccharides from Astragalus membranaceus (Fisch) Bge. var. Mongholicus (Bge.) Hsiao roots under ultrasound-assisted conditions. In this study, 16 different natural deep eutectic solvents were designed to screen the best solvent for extracting Astragalus polysaccharides (APSs). Based on the yield and recovery of APSs, a natural deep eutectic solvents composed of choline chloride and oxalic acid with a molar ratio of 1:2 was selected. The related factors affecting polysaccharides extraction and solvent precipitation were investigated. To improve the operating methodology, single-factor trials, a Plackett-Burman design, and a Box-Behnken design were used. The optimal extraction process conditions were obtained as follows: water content of 55%, liquid-solid ratio of 24 mL/g, ultrasonic irradiation time of 54 min, ultrasonic irradiation temperature of 50 °C, ultrasonic irradiation power of 480 W, ethanol precipitation time of 24 h, and ethanol concentration of 75%. Under optimal extraction conditions, the recovery of APSs was 61.4 ± 0.6 mg/g. Considering the special matrix characteristics of A. membranaceus var. Mongholicus roots, physical-technology-based ultrasonic waves promote penetration, and the mass transfer function also solves the bottleneck of high-viscosity deep eutectic solvents in the extraction stage. In comparison with the conventional method, the proposed method based on deep eutectic solvents isolation can significantly increase APSs recovery, which is beneficial to simplifying the process of polysaccharides purification by using solvent properties to separate extracts and reduce impurities in APSs.
Assuntos
Astragalus propinquus , Solventes Eutéticos Profundos , Solventes , Água , Etanol , Polissacarídeos , Extratos VegetaisRESUMO
BACKGROUND: Traditional Chinese medicine (TCM) has been widely recognized and accepted worldwide to provide favorable therapeutic effects for cancer patients. As Andrographis paniculata has an anti-tumor effect, it might inhibit lung cancer. OBJECTIVE: The drug targets and related pathways involved in the action of Andrographis paniculata against lung cancer were predicted using network pharmacology, and its mechanism was further explored at the molecular level. METHODS: This work selected the effective components and targets of Andrographis paniculata against the Traditional Chinese Medicine System Pharmacology (TCMSP) database. Targets related to lung cancer were searched for in the GEO database (accession number GSE136043). The volcanic and thermal maps of differential expression genes were produced using the software R. Then, the target genes were analyzed by GO and KEGG analysis using the software R. This also utilized the AutoDock tool to study the molecular docking of the active component structures downloaded from the PubChem database and the key target structures downloaded from the PDB database, and the docking results were visualized using the software PyMol. RESULTS: The results of molecular docking show that wogonin, Mono-O-methylwightin, Deoxycamptothecine, andrographidine F_qt, Quercetin tetramethyl (3',4',5,7) ether, 14-deoxyandrographolide, andrographolide-19-ß-D-glucoside_qt and 14-deoxy-11-oxo-andrographolide were potential active components, while AKT1, MAPK14, RELA and NCOA1 were key targets. CONCLUSION: This study showed the main candidate components, targets, and pathways involved in the action of Andrographis paniculata against lung cancer.
Assuntos
Andrographis paniculata , Neoplasias Pulmonares , Simulação de Acoplamento Molecular , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Medicina Tradicional Chinesa , Farmacologia em RedeRESUMO
Premature ovarian failure (POF) is defined as lost ovarian functions before the age of 40. Three possible molecular markers (PLA2G4A, miR-29a, and miR-144) have been identified in our previous study by integrated analysis of mRNA and miRNA expression profiles. The present study aimed to evaluate American ginseng root's protective potential against POF by studying transcriptional and protein variations between American ginseng treatments and controls in rats. 4-Vinylcyclohexene diepoxide (VCD) was administered to rats for 14 days to induce POF. Additionally, American ginseng was administered to POF rats for one month, and PLA2G4A, miR-29a, and miR-144 expressions were measured in rat ovaries by qRT-PCR. PLA2G4A protein expression was examined by Western Blot, and PGE2, LH, FSH, and E2 serum levels were detected by ELISA. PLA2G4A mRNA and protein were downregulated in American ginseng-treated rats, miR-29a and miR-144 levels increased, and PGE2 serum levels decreased, while LH, FSH, and E2 increased compared to POF induction alone. Analysis of transcriptional and protein variations suggested that American ginseng protects the ovary against POF by regulating prostaglandin biosynthesis, ovulation, and preventing ovarian aging. High hormone levels (PGE2, FSH, and LH) were reduced, and E2 secretion approached normal levels, leading to improved POF symptoms and abnormal ovulation.
Assuntos
Panax/química , Extratos Vegetais/administração & dosagem , Insuficiência Ovariana Primária/tratamento farmacológico , Antígeno 12E7 , Animais , Antígenos CD/sangue , Moléculas de Adesão Celular/sangue , Cicloexenos/toxicidade , Dinoprostona/sangue , Feminino , Hormônio Foliculoestimulante Humano/sangue , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hormônio Luteinizante/sangue , Extratos Vegetais/química , Insuficiência Ovariana Primária/sangue , Insuficiência Ovariana Primária/induzido quimicamente , Insuficiência Ovariana Primária/patologia , Ratos , Compostos de Vinila/toxicidadeRESUMO
Preclinical Research Premature ovarian failure (POF) is defined by the WHO as the loss of physiological ovarian function before the age of 40. The effect of American ginseng and its underlying mechanisms in preventing and treating premature ovarian failure (POF) was studied in female Sprague-Dawley rats where POF was induced by ip administration of 4-vinylcyclohexene diepoxide (VCD). Rat behavior, serum hormone levels, ovarian and uterine size, pathological features, and ovarian tissue expression of genes associated with POF were assessed in controls, untreated POF model rats, and POF model rats treated with low- (1.125 g/kg), medium- (2.25 g/kg), and high-dose (4.5 g/kg) American ginseng. Compared with untreated POF model rats, those treated with medium- and high-dose American ginseng had more stable behavior and better coat appearance as well as serum hormone levels closer to those in control rats. Moreover, treatment with medium- or high-dose American ginseng increased ovarian and uterine size. Hematoxylin and eosin-staining revealed mature follicles and endometrium with an alternating concave/convex surface structure with visible capillaries and glands in ginseng- treated POF rats. PLA2G4A expression was positively correlated with POF, while the expression levels of PAPPA, STC2, CCL2, and NELL1 were negatively correlated with POF. Our study showed that American ginseng may effectively prevent POF and alleviate POF symptoms by regulating serum hormone levels and altering the expression levels of genes related to POF in ovarian tissue.