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In this paper, the extraction rate of crude polysaccharides and the yield of polysaccharides from Hippocampus served as test indicators. The comprehensive evaluation indicators were assigned by the R language combined with the entropy weight method. The Box-Behnken design-response surface methodology(BBD-RSM) and the deep neural network(DNN) were employed to screen the optimal parameters for the polysaccharide extraction from Hippocampus. These two modeling methods were compared and verified experimentally for the process optimization. This study provides a reference for the industrialization of effective component extraction from Chinese medicinals and achieves the effective combination of modern technology and traditional Chinese medicine.
Assuntos
Carboidratos da Dieta , Polissacarídeos , Hipocampo , Redes Neurais de Computação , TemperaturaRESUMO
Irritable bowel syndrome (IBS) is a prevalent gastrointestinal dysfunction. Cimifugin is an active component of Radix saposhnikoviae which is effective for maintaining intestinal barrier integrity and intestinal function. This study aimed to investigate the treatment efficacy of Cimifugin on intestinal barrier dysfunction and to unveil the relevant mechanism through network pharmacology and experimental verification as well as molecular docking. Through SuperPred and Pubchem databases, the targets of Cimifugin were obtained. The disease targets were screened using Disgenet and GEO databases. With STRING database and Cytoscape software, the analysis of PPI network was performed. In DAVID database, the hub genes of Cimifugin were analyzed using GO and Pathway enrichment analyses. To validate the binding of Cimifugin with core targets, molecular docking was performed. The in vitro cellular model of intestinal barrier was established via the induction of Caco2 cells with LPS. TEER was used to detect epithelial barrier function and permeability was measured using FITC-dextran (FD4). Western blotting was used to measure the expressions of SIRT1, tight junction proteins, and NRF2/HO-1 signaling pathway-related proteins. The fluorescence intensity of ZO-1, Occludin, and Claudin-1 was detected using immunofluorescence staining. ELISA was used to detect the expression levels of inflammatory cytokines. Through the integration of all targets of IBS and Cimifugin, 94 frequent drug-disease-related targets were identified. These targets were enriched in some signaling pathways, like cellular responses to stress, cellular responses to stimuli, and VEGFA-VEGFR2. Ten hub genes including PTGS2, ANPRP, TGFB1, ACACA, SIRT1, NEF2L2, APEX1, IL6, AKT1, and HSP90AB1 were obtained. Cimifugin showed strong affinity with four key genes, including AKT1, SIRT1, IL6, and NFE2L2 (NRF2), which were obtained through the intersection of hug genes with cellular responses to stimuli. In vitro experiments showed that Cimifugin ameliorated LPS-induced intestinal barrier injury in Caco2 cells via upregulating SIRT1 to modulate NRF2/HO-1 signaling pathway. Cimifugin could alleviate intestinal barrier dysfunction in IBS by upregulating SIRT1 to regulate the NRF2/HO-1 signaling pathway.
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ETHNOPHARMACOLOGICAL RELEVANCE: In China, Hordei Fructus Germinatus (HFG) is the germinated and dried fruit of Hordeum vulgare L, which is commonly used in clinical Chinese medicine. Traditional Chinese Medicine (TCM) theory holds that HFG can be both medicinal and edible, which means that it is derived from food medicine. Raw HFG and roasted HFG are used to treat hypogalactia, hyperprolactinemia and indigestion. In recent years, the lactogenic and galactophygous effects of HFG have attracted increasing attention. Nevertheless, there is much confusion over the use of raw and processed HFG, and the mechanism of its lactogenic effect seems remains poorly understood. AIM OF THE STUDY: This study aimed to explore the lactogenic effect of raw HFG and roasted HFG on rats with overloaded lactation and to reveal the underlying molecular mechanism. MATERIALS AND METHODS: Raw and processed HFG water decoctions were given to overloaded lactation model rats at a dose of 1.7800 g kg-1·d-1, and the control group was given the same volume of water. The lactogenic effect of raw and processed HFG was evaluated by measuring daily lactation, body weight and pup body weight, serum PRL, E2, and GH contents after parturition, and the pathological characteristics of mammary tissue sections. cDNA microarrays can be used to screen diverse gene expression patterns and signaling pathways related to prolactin. The expression of relevant differentially expressed genes was verified by real-time PCR and western blotting. RESULTS: In vivo experiments demonstrated that the raw HFG water decoction stimulated mammogenesis, accelerated the transformation of the lobular acinar system, resulted in denser mammary epithelial cells and thicker glandular ducts that were full of milk and facilitated the secretion of milk. Moreover, HFG increased PRL, E2, and GH levels, pup body weight, daily lactation and the body weight of lactating rats. Following gene chip identification, KEGG pathway enrichment analysis revealed genes that were highly related to prolactin in the prolactin signaling pathway and JAK-STAT signaling pathway, and the main differentially expressed genes were Jak2 (down), Stat5α (up), cyclin D1 (up), SOCS1 (up), CISH (down) and PRLR (up). Compared with the control group, RT-PCR results indicated that Jak2 and CISH were downregulated and that Stat5α, cyclin D1, SOCS1 and PRLR were upregulated. Western blot assays showed that PRLR, STAT5α and cyclin D1 levels in the mammary glands of the raw HFG water decoction group were significantly increased, which was consistent with the results of cDNA microarray screening. CONCLUSION: The present study reveals that raw HFG effectively enhances lactation in rats, possibly by influencing the prolactin/JAK-STAT signaling pathway.