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•Deep eutectic solvents (DESs) can simplify the purification process of polysaccharides.•Taking full advantage of the high density and viscosity characteristics of DESs.•Proposed ...method promotes solvent infiltration for high-efficiency extraction.•Proposed method effectively guarantees the yield and purity of polysaccharides.•Proposed method is more consistent with the principles of green and precise extraction.
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.
We previously screened 6 differentially expressed miRNAs in ovarian tissues of 4-vinylcyclohexene diepoxide (VCD)-treated premature ovarian failure (POF) model in SD rats, including miRNA-190a-5p, ...miRNA-98-5p, miRNA-29a-3p, miRNA-144-5p, miRNA-27b-3p, miRNA-151-5p. In this study, to investigate the mechanisms causing the onset of POF, we first identified miRNAs with earlier differential expression at consecutive time points in the VCD-treated rat POF model and explored the mechanisms by which the target miRNAs promote POF. The SD rats were injected with VCD for 15 days to induce POF. Additionally, we collected rat blood and ovaries at the same time every day for 15 consecutive days, and luteinizing hormone (LH), follicle-stimulating hormone (FSH), Anti-Mullerian hormone (AMH), and estradiol (E
2
) serum levels were detected by ELISA. Six miRNAs expression were measured in rat ovaries by qRT-PCR. Dual-luciferase reporter gene assays were employed to predict and verify the target gene (PHLPP1) of target miRNAs (miRNA-190a-5p). Western blot was examined to detect the expression levels of PHLPP1, AKT, p-AKT, FOXO3a, p-FOXO3a, and LHR proteins on the target gene PHLPP1 and its participation in the primordial follicular hyperactivation-related pathways (AKT-FOXO3a and AKT-LH/LHR). During the VCD modeling POF rat ovaries, miRNA-190a-5p was the first to show significant differential expression, i.e., 6th of VCD treating, and PHLPP1 was verified to be a direct downstream target of it. Starting from the 6th of VCD treatment, the more significant the up-regulation trend of miRNA-190a-5p expression, the more obvious the down-regulation trend of PHLPP1 and LHR mRNA and protein expression, accompanied by the more severe phosphorylation of AKT and FOXO3a proteins, thus continuously over-activating the rat primordial follicle to promote the development of POF. In conclusion, miRNA-190a-5p may become a potential biomarker for early screening of POF, and it can continuously activate primordial follicles in rats by targeting the expression of PHLPP1 and key proteins in the AKT-FOXO3a and AKT-LH/LHR pathways.
Molecular characterization of insulin resistance, a growing health issue worldwide, will help to develop novel strategies and accurate biomarkers for disease diagnosis and treatment.
Integrative ...analysis of gene expression profiling and gene regulatory network was exploited to identify potential biomarkers early in the development of insulin resistance.
RNA was isolated from livers of animals at three weeks of age, and whole-genome expression profiling was performed and analyzed with Agilent mouse 4×44K microarrays. Differentially expressed genes were subsequently validated by qRT-PCR. Functional characterizations of genes and their interactions were performed by Gene Ontology (GO) analysis and gene regulatory network (GRN) analysis.
A total of 197 genes were found to be differentially expressed by fold change ≥2 and P < 0.05 in BKS-db +/+ mice relative to sex and age-matched controls. Functional analysis suggested that these differentially expressed genes were enriched in the regulation of phosphorylation and generation of precursor metabolites which are closely associated with insulin resistance. Then a gene regulatory network associated with insulin resistance (IRGRN) was constructed by integration of these differentially expressed genes and known human protein-protein interaction network. The principal component analysis demonstrated that 67 genes in IRGRN could clearly distinguish insulin resistance from the non-disease state. Some of these candidate genes were further experimentally validated by qRT-PCR, highlighting the predictive role as biomarkers in insulin resistance.
Our study provides new insight into the pathogenesis and treatment of insulin resistance and also reveals potential novel molecular targets and diagnostic biomarkers for insulin resistance.
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.
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.
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.
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-β-D-glucoside_qt and 14-deoxy-11-oxo-andrographolide were potential active components, while AKT1, MAPK14, RELA and NCOA1 were key targets.
This study showed the main candidate components, targets, and pathways involved in the action of Andrographis paniculata against lung cancer.
Deep eutectic solvents (DESs) are receiving increased attention from researchers in many areas of science and technology. During the actual operation, the high viscosity of DESs, especially for ...solid-phase sample application, can be diminished by combination of water in the solvent system, heating, and consuming a large volume of DESs. In addition, plant matrix properties of Schisandra chinensis Baill fruits should be considered including sugars-rich and full of stone cells, which also dissolve into the solvent and to reduce extraction capacity. In this study, an ultrasonic probe-based system with a direct delivery of ultrasound to sample was developed to help the matrix materials become hydrated and swelled, which increases the diffusion and flow of solvent into the dry matrix and increases mass transfer, and also minish ultrasonic energy loss. Some parameters that potentially influence circulating and pulsating ultrasonic probe extraction (CPUPE) were methodically examined and improved using the Plackett–Burman, Box–Behnken design, and the optimum practical parameters were as follows: water content of DES, 20.2%; liquid-to-solid ratio, 11.0 mL/g; total ultrasonic treatment time, 24.7 min; ultrasonic setting temperature, 323 K; ultrasonic irradiation power, 90 W; and circulating rate, 25 mL/min. CPUPE provided a accessibility to use substituted green solvents of DESs (choline chloride-lactic acid) by improving their extraction performance. The high sample extraction capacity and circulation function of the suggested CPUPE technique makes it a quick and effective method for complex substrate plant samples. Under the circulating system, the osmotic pressure difference of the solvent is significantly increased. The CPUPE make the kind method operation simple, fast, can be apply in the on-line production analysis.
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•CPUPE is suitable for high viscosity solvent, especially for DES.•CPUPE overcomes many problems brought from DES aqueous solution.•CPUPE effectively guarantee big sample amounts for ultrasonic probe application.•CPUPE promote solvent infiltration ability for the high-efficiency extraction.•CPUPE effectively guarantees the quality and yield of lignans.
Abstract
Premature ovarian failure (POF) contributes to amenorrhoea, infertility, early onset of menostasia and osteoporosis. This study profiled differentially expressed miRNAs for association with ...POF development. Ovarian tissue samples from 4-vinylcyclohexene diepoxide (VCD)-induced rat POF and normal rats were profiled for differentially expressed miRNAs using miRNA microarrays. A total of 63 miRNAs were up-regulated and 20 miRNAs were down-regulated in rat POF tissues versus the control tissues. qRT-PCR verified some of these altered miRNAs, i.e. miR-29a and miR-144 were down-regulated in POF tissues, which may target expression of PLA2G4A that is involved in prostaglandin biosynthesis, whereas miR-27b and miR-190 were up-regulated in POF tissues by negative control of PAPPA and CCL2 expression, respectively, both of which have been shown to relate to response to hormone stimulus. Moreover, the up-regulated miR-151 and miR-672 can also target expression of TNFSF10 and FNDC1, which have been shown to positively regulate cell apoptosis. Profiling of differentially expressed miRNAs in POF provided a novel insight into the molecular events involving the role of miRNAs in POF development with specific emphasis upon miR-27b, miR-190, miR-151, miR-672, miR-29a and miR-144.
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.
Diabetes mellitus is one of the primary diseases that pose a threat to human health. The focus of the present study is type II diabetes (T2D), which is caused by obesity and is the most prevalent ...type of diabetes. However, genome-scale transcriptional analysis of diabetic liver in the development process of T2D is yet to be further elucidated. Microassays were performed on liver tissue samples from three-, six- and nine-week-old
mice with diabetes and
mice to investigate differentially expressed mRNA. Based on the results of genome-scale transcriptional analysis, five genes were screened in the present study: chromobox 8 (
), de-etiolated homolog 1 and damage specific DNA binding protein 1 associated 1 (
), Phosphoinositide-3-kinase regulatory subunit 6 (
), WD repeat domain 41 (
) and Glycine Amidinotransferase (
). At three weeks of age, no significant differences in levels or ratios of expression were observed. However, at six and nine weeks, expression of
,
,
and
was significantly upregulated (P<0.05) in the
model group compared with the control group, whereas
expression was significantly downregulated (P<0.05). These results suggest that T2D-related differential expression of genes becomes more marked with age, which was confirmed via reverse transcription-quantitative polymerase chain reaction. Genome-scale transcriptional analysis in diabetic mice provided a novel insight into the molecular. events associated with the role of mRNAs in T2D development, with specific emphasis upon
,
,
,
and
. The results of the present study may provide rationale for the investigation of the target genes of these mRNAs in future studies.
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.
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