The cell wall polysaccharides were extracted from Sparassis latifolia fruit bodies by acid–alkali and superfine-grinding assisted methods, and the chemical characterization and in vitro immunity activities of these polysaccharide fractions were studied and compared. Results showed that superfine-grinding assisted extraction exhibited the highest yield of polysaccharides (SP, 20.80%) and low β-glucan content (19.35%) compared with alkaline extracts. The results revealed that the 20% ethanol precipitated fraction (20E) from SP was mainly composed of β-(1→3)-glucan and α-(1→4)-glucan. With the increase of ethanol precipitation, the fractions (30E, 40E, 50E) were identified as α-(1→4)-glucan with different molecular weights and conformations. Cell wall polysaccharides extracted through NaOH (NSP) and KOH (KSP) extraction had similar yields with 8.90% and 8.83%, respectively. Structural analysis indicated that the purified fraction from KSP (KSP-30E) was a β-(1→3)-glucan backbone branched with β-(1→6)-Glcp, while the purified fraction from NSP (NSP-30E) mainly contained β-(1→3)-glucan with a small number of α-linked-Glcp. The two fractions both exhibited rigid chain conformation in aqueous solutions. All polysaccharide fractions exerted the activity of activating Dectin-1 receptor in vitro, and the KSP-30E mainly identified as β-(1→3)-glucan with the terminal group via 1→6-linkage attached at every third residue exhibited a stronger enhancing effect than other fractions. Results suggested that KOH extraction could be efficient for the preparation of bioactive β-(1→3, 1→6)-glucan as a food ingredient.

Polysaccharides with molecular weights ranging from 1.75 × 10³ to 1.14 × 10⁴ g/mol were obtained from the fruit bodies of Ganoderma lucidum. The multiple fingerprints and macrophage immunostimulatory activity of these fractions were analyzed as well as the fingerprint–activity relationship. The correlation analysis of molecular weight and immune activity demonstrated that polysaccharides with molecular weights of 4.27 × 10³~5.27 × 10³ and 1 × 10⁴~1.14 × 10⁴ g/mol were the main active fractions. Moreover, the results showed that galactose, mannose, and glucuronic acid were positively related to immunostimulatory activity. Additionally, partial least-squares regression and grey correlation degree analyses indicated that three peaks (P2, P3, P8) in the oligosaccharide fragment fingerprint significantly affected the immune activity of the polysaccharides. Hence, these ingredients associated with activity could be considered as markers to assess Ganoderma lucidum polysaccharides and their related products, and the study also provides a reference for research on the spectrum–effect relationship of polysaccharides in the future.

Background: Centella asiatica (CA) has been used to address cancer for centuries in traditional Chinese medicine (TCM). Previous studies demonstrated its anti-angiogenesis efficacy, but the underlying mechanism of its action remains to be further clarified. This study aims to investigate the underlying mechanisms of CA and its triterpenes in anti-angiogenesis for cancer therapeutics through network pharmacology and experimental validation. Methods: Cytoscape was used to construct a network of compound–disease targets and protein–protein interactions (PPIs) from which core targets were identified. GO and KEGG analyses were performed using Metascape, and the AutoDock-Vina program was used to realize molecular docking for further verification. Then, VEGF165 was employed to establish an induced angiogenesis model. The anti-angiogenic effects of CA were evaluated through assays measuring cell proliferation, migration, and tubular structure formation. Results: Twenty-five active ingredients in CA had potential targets for anti-angiogenesis including madecassoside, asiaticoside, madecassic acid, asiatic acid, and asiaticoside B. In total, 138 potential targets for CA were identified, with 19 core targets, including STAT3, SRC, MAPK1, and AKT1. A KEGG analysis showed that CA is implicated in cancer-related pathways, specifically PD-1 and AGE-RAGE. Molecular docking verified that the active components of CA have good binding energy with the first four important targets of angiogenesis. In experimental validation, the extracts and triterpenes of CA improved VEGF165-induced angiogenesis by reducing the proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs). Conclusions: Our results initially demonstrate the effective components and great anti-angiogenic activity of CA. Evidence of the satisfactory anti-angiogenic action of the extracts and triterpenes from CA was verified, suggesting CA’s significant potential as a prospective agent for the therapy of cancer.

Background: Madecassoside is widely utilized in wound healing due to its multiple physiological activities. However, its limited bioavailability and solubility hinder its clinical application. Enzymatic hydrolysis has been employed to enhance the bioavailability and bioactivity of natural products, but its potential for modifying madecassoside remains unexplored. Methods: In this study, we prepared MA1G and MA2G through enzymatic hydrolysis, inspired by the metabolic processes of madecassoside. Network pharmacology was employed to investigate the mechanisms of these madecassoside derivatives (MDs) in wound healing, and molecular docking was performed to evaluate their binding affinities. Transdermal permeation studies, scratch assays, and antioxidant and anti-inflammatory tests were conducted to characterize the biological properties and activities of MDs. Results: Network pharmacology identified TLR4, NF-κB, and STAT3 as key targets for wound healing, and the MDs inhibited the expression of these proteins in vitro. Additionally, the results demonstrated that MDs exhibited robust reactive oxygen species (ROS) scavenging activity (43.05–147.50% reduction) and significantly enhanced cell migration (36.76–77.28% increase). Notably, the biomodified MA2G showed superior transdermal permeability and biological activities. Conclusions: This paper represents the first report directly comparing the biological activities of the parent compound (madecassoside) and its metabolites while simultaneously proposing a novel therapeutic strategy for wound healing.

Background/Objectives: Atopic dermatitis (AD) is a chronic inflammatory skin disorder that has attracted global attention, and alkaloids from Plumula Nelumbinis have been shown to have anti-inflammatory activity. Fermentation has been used for the structural modification of natural compounds to improve bioavailability and activity, but the AD therapeutic efficacy and mechanism of the fermented Plumula Nelumbinis (FPN) are still unclear. Methods: The potential targets of FPN for AD were preliminarily screened using network pharmacology, and then PCR and WB were used to prove the therapeutic effect of FPN in AD. Results: Network pharmacology indicated that mTOR and Jun were key targets for AD. The experiments in vitro showed that FPN could effectively block AKT/mTOR and AKT/Jun-mediated inflammatory signaling pathways. Moreover, FPN can also alleviate SDS-induced inflammation in zebrafish. It is also found that the anti-inflammatory activity of Plumula Nelumbinis was enhanced by Aspergillus oryzae fermentation, and the oil phase of the fermentation product showed better activity, which may be due to microbial fermentation changing the structure of the original alkaloids. Conclusions: This study elucidated the potential mechanisms of alkaloids derived from fermented Plumula Nelumbinis against AD; it may also provide a scientific basis for the development of new drugs for AD.

Rosacea is a chronic inflammatory skin disease and is usually accompanied by extensive macrophage infiltration. There is growing evidence suggesting that neurotransmitter 5-hydroxytryptamine (5-HT) plays a crucial role in inflammatory reactions. However, the interaction between 5-HT and rosacea is still unclear. Here, we hypothesized that the inflammation of rosacea is partly caused by 5-HT, and we investigated the underlying mechanism. In this study, we employed a rosacea model induced by LL-37, which is usually applicated as a rosacea stimulator, to investigate the effects of 5-HT on rosacea in vitro and in vivo. In LL-37-(4 μM)-induced THP-1-derived macrophages, 5-HT (400 μM) further promoted the secretion of inflammatory cytokines and polarized macrophages towards M1 phenotype, which could promote an inflammatory response. Further research revealed that exposure to LL-37 and 5-HT (L5) selectively upregulated HTR3A mRNA expression but not HTR2A or HTR7 and induced colocalization of 5-HT with HTR3A. Notably, application of antagonist tropisetron (TPS) and siRNA of HTR3A suppressed L5-induced inflammation. Meanwhile, 5-HT (25 μg each injection a total of three times) deteriorated skin erythema, stimulated dermal inflammatory cell infiltration, and promoted the secretion of inflammatory cytokines in LL-37 (40 μL and 320 μM each injection a total of four times) induced rosacea-like mice, while these undesirable effects were reversed by using TPS. Our findings revealed that neurotransmitter 5-HT further promoted LL-37-induced rosacea-like inflammation through HTR3A. Our study highlights HTR3A as a promising therapeutic target, which warrants further in-depth investigation into its clinical applicability.