In this study, the Box–Behnken Design (BBD) was adopted to optimize the ultrasound-assisted extraction (UAE) conditions of polyphenols from Hericium erinaceus (H. erinaceus) on the basis of single-factor experiments, with extraction time, solid–liquid ratio and ethanol concentration as the key investigation factors. The optimal extraction parameters were determined as follows: extraction time of 56.85 min, solid–liquid ratio of 1:56.71 g/mL and ethanol concentration of 44.64%, under which the actual yield of the total polyphenol crude extract (TPCE) reached 0.9985 ± 0.03%, which was highly consistent with the theoretical predicted value of 0.9960%, verifying the good fitting degree of the established model. Taking L-ascorbic acid as the positive control, the antioxidant activity of TPCE was evaluated by determining its scavenging capacity against ABTS·⁺, ·OH and DPPH· free radicals, and the half-maximal effective concentration (EC₅₀) values were measured to be 0.8850, 0.9490 and 4.198 mg/mL, respectively. With acarbose as the reference drug, the inhibitory effects of TPCE on α-amylase and α-glucosidase related to carbohydrate metabolism were assayed, and the corresponding half-maximal inhibitory concentration (IC₅₀) values were 0.0135 and 130.3 mg/mL, respectively. Furthermore, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was employed for the tentative identification of bioactive components in TPCE, and a total of 48 and 64 chemical constituents were characterized in negative and positive ion modes, respectively, providing a chemical basis for the biological activities of TPCE. This study confirmed that UAE is an efficient and feasible technology for extracting polyphenols from H. erinaceus, which lays a theoretical foundation for the development and utilization of its polyphenols, and also provides novel insights into the development of natural functional ingredients and potential therapeutic agents for the intervention of type 2 diabetes. Additionally, the findings further validate edible fungi as a valuable reservoir of natural bioactive substances, with promising application prospects in the research and development of functional foods and pharmaceuticals targeting metabolic diseases.

α-Amylase and α-glucosidase are complementary therapeutic targets in type 2 diabetes; simultaneous inhibition provides a dual-blockade strategy to suppress postprandial hyperglycemia. Dipyridamole, a conventional antiplatelet drug with proven anti-inflammatory, antioxidant, and antiproliferative effects and a favorable safety profile, is a candidate for repurposing. This study assessed in vitro antioxidant activity and inhibitory actions against both enzymes, and employed molecular docking, molecular dynamics simulations to elucidate mechanisms. Dipyridamole showed EC₅₀ values of 589.7 μM (ABTS+·), 652.2 μM (·OH), and 1607.1 μM (DPPH·), and IC₅₀ values of 257.6 μM (α-amylase) and 348.8 μM (α-glucosidase). Molecular docking yielded binding free energies of −6.5 and −6.0 kcal/mol for α-amylase and α-glucosidase, respectively, with binding mediated mainly by hydrogen bonds and hydrophobic interactions. Molecular dynamics simulations revealed that, under the present simulation conditions, both the apo forms of α-amylase and α-glucosidase and their respective dipyridamole complexes exhibited relatively stable dynamical behavior throughout the course of the trajectories. Overall, dipyridamole showed not only moderate free radical scavenging capacity but also inhibitory activity against α-amylase and α-glucosidase, suggesting that this molecule may serve as a promising lead compound for further relevant drug development and functional exploration.

Background: Inonotus obliquus (Chaga), a medicinal and edible macrofungus abundant in bioactive polyphenols, is a potential source of natural antioxidants and anti-inflammatory agents for functional foods. This study aimed to evaluate the antioxidant capacity of three key polyphenols (osmundacetone [OS], protocatechuic aldehyde [PAH], protocatechuic acid [PA]) from I. obliquus and decipher their anti-inflammatory mechanisms via the MyD88/TLR4/NF-κB pathway in a gout-related model. Methods: Antioxidant activity was assessed by xanthine oxidase (XO) inhibition (IC₅₀), superoxide anion (O₂⁻) scavenging, and structure–activity relationship (SAR) analysis; in a monosodium urate (MSU)-induced acute gout cell model, reactive oxygen species (ROS), nitric oxide (NO), lactate dehydrogenase (LDH), superoxide dismutase (SOD), pro-inflammatory cytokines (TNF-α, IL-1β) were quantified, and MyD88/TLR4/NF-κB pathway proteins were analyzed by Western blot. Results: OS showed the strongest XO inhibition (IC₅₀ = 4.91 mM), followed by PAH (IC₅₀ = 5.92 mM) and PA (IC₅₀ = 26.53 mM); OS exerted dual redox effects by scavenging O₂⁻ and suppressing XO-mediated O₂⁻ generation, with its conjugated C=C-carbonyl system and PAH’s aldehyde group enhancing XO binding. All polyphenols and I. obliquus crude extract significantly reduced ROS, NO, LDH, and cytokines (p < 0.05), increased SOD, and downregulated TLR4, MyD88, and NF-κB expression. Conclusions: I. obliquus-derived polyphenols exhibit obvious antioxidant and xanthine oxidase inhibitory effects, and regulate oxidative stress, pro-inflammatory mediators, and the MyD88/TLR4/NF-κB signaling pathway in monosodium urate-stimulated RAW 264.7 inflammatory macrophages, supporting their development as natural functional food ingredients and potential candidates for gout-related and oxidative stress-associated inflammatory cellular disorders.