Bioactive triterpenoid compounds of Poria cocos (Schw.) Wolf in the treatment of diabetic ulcers via regulating the PI3K-AKT signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic ulcers represent a chronic condition characterized by prolonged hyperglycemia and delayed wound healing, accompanied by endocrine disorders, inflammatory responses, and microvascular damage in the epidermal tissue, demanding effective clinical treatment approaches. For thousands of years, ancient Chinese ethnopharmacological studies have documented the use of Poria cocos (Schw.) Wolf in treating diabetic ulcers. Recent research has substantiated the diverse pharmacological effects of Poria cocos (Schw.) Wolf, including its potential to alleviate hyperglycemia and exhibit anti-inflammatory, antioxidant, and immune regulatory properties, which could effectively mitigate diabetic ulcer symptoms. Furthermore, being a natural medicine, Poria cocos (Schw.) Wolf has demonstrated promising therapeutic effects and safety in the management of diabetic ulcers, holding significant clinical value. Despite its potential clinical efficacy and applications in diabetic ulcer treatment, the primary active components and underlying pharmacological mechanisms of Poria cocos (Schw.) Wolf remains unclear. Further investigations are imperative to establish a solid foundation for drug development in this domain. AIM OF THE STUDY AND MATERIALS AND METHODS: In this study, we aimed to identify the active compounds and potential targets of Poria cocos (Schw.) Wolf using UHPLC-Q-TOF-MS and TCMSP databases. Additionally, we attempt to identify targets related to diabetic ulcers. Following enrichment analysis, a network of protein-protein interactions was constructed to identify hub genes based on the common elements between the two datasets. To gain insights into the binding activities of the hub genes and active ingredients, molecular docking analysis was employed. Furthermore, to further validate the therapeutic effect of Poria cocos (Schw.) Wolf, we exerted in vitro experiments using human umbilical vein vascular endothelial cells and human myeloid leukemia monocytes (THP-1). The active ingredient of Poria cocos (Schw.) Wolf was applied in these experiments. Our investigations included various assays, such as CCK-8, scratch test, immunofluorescence, western blotting, RT-PCR, and flow cytometry, to explore the potential of Poria cocos (Schw.) Wolf triterpenoid extract (PTE) in treating diabetic ulcers. RESULTS: The findings here highlighted PTE as the primary active ingredient in Poria cocos (Schw.) Wolf. Utilizing network pharmacology, we identified 74 potential targets associated with diabetic ulcer treatment for Poria cocos (Schw.) Wolf, with five hub genes (JUN, MAPK1, STAT3, AKT1, and CTNNB1). Enrichment analysis revealed the involvement of multiple pathways in the therapeutic process, with the PI3K-AKT signaling pathway showing significant enrichment. Through molecular docking, we discovered that relevant targets within this pathway exhibited strong binding with the active components of Poria cocos (Schw.) Wolf. In vitro experiments unveiled that PTE (10 mg/L) facilitated the migration of human umbilical vein vascular endothelial cells (P < 0.05). PTE also increased the expression of CD31 and VEGF mRNA (P < 0.05) while activating the expressions of p-PI3K and p-AKT (P < 0.05). Moreover, PTE demonstrated its potential by reducing the expression of IL-1ß, IL-6, TNF-α, and NF-κB mRNA in THP-1 (P < 0.05) and fostering M2 macrophage polarization. These results signify the potential therapeutic effects of PTE in treating diabetic ulcers, with its beneficial actions mediated through the PI3K-AKT signaling pathway. CONCLUSIONS: PTE is the main active ingredient in Poria cocos (Schw.) Wolf that exerts therapeutic effects. Through PI3K-AKT signaling pathway activation and inflammatory response reduction, PTE promotes angiogenesis, thereby healing diabetic ulcers.

Nanostructured lipid carriers (NLCs) stabilized by natural or synthetic emulsifiers for lutein delivery: Improved physicochemical stability, antioxidant activity, and bioaccessibility.

This study investigated the impacts of individual emulsifiers on the physicochemical stability, antioxidant ability, and in vitro digestion behavior of lutein-loaded nanostructured lipid carriers (NLCs). NLCs particles stabilized by ethyl lauroyl arginate, rhamnolipid, or tea saponin were fabricated by high-pressure microfluidization method. Differential scanning calorimetry and X-ray diffraction results confirmed the regulatory effect of emulsifiers on the crystallization behavior of NLCs. NLCs stabilized by rhamnolipid presented higher encapsulation efficiency (94.73%) for lutein than those stabilized by tea saponin (90.39%) or ethyl lauroyl arginate (88.86%). Meanwhile, the stability of embedded lutein during storage or photothermal treatments was greatly enhanced. Individual emulsifiers, together with lutein, endowed NLCs with excellent antioxidant capacity. During in vitro digestion, rhamnolipid-stabilized NLCs showed the slowest release of free fatty acids (50.87%) and provided an optimal sustained release for lutein with relatively high bioaccessibility (23.01%).

Effects of microfluidization and thermal treatment on the characterization and digestion of curcumin loaded protein-polysaccharide-tea saponin complex nanoparticles.

Microfluidization (50-150 MPa) and thermal treatment (45-85 °C) were applied to modulate the physicochemical stability, molecular interaction and microstructure of zein-proplyene glycol alginate (PGA)-tea saponin (TS) complex nanoparticles for delivery of curcumin. The size of these complex nanoparticles was decreased from 583.1 to 267.4 nm as the microfluidization pressure was increased from 0 to 100 MPa. In the combined treatment of microfluidization and heating, 100 MPa and 75 °C were the optimum parameters to prepare zein-PGA-TS complex nanoparticles for a better protection of curcumin against various environmental stresses. SEM revealed a synergistic effect of microfluidization and heating on the fabrication of complex nanoparticles with a more uniform size and spherical shape. During in vitro gastrointestinal digestion, the complex nanoparticles showed an excellent gastric stability and a sustained release of curcumin in the small intestinal phase. These findings interpreted the effects of microfluidization and thermal treatment on the functional properties of protein-polysaccharide-surfactant complex nanoparticles that can be utilized to develop food grade nanoparticles with enhanced stability and controllable digestion behaviour.

Fabrication, structural characterization and functional attributes of polysaccharide-surfactant-protein ternary complexes for delivery of curcumin.

In this study, the nanocomplexes as a novel delivery system for curcumin, were successfully fabricated using high methoxyl pectin (HMP), individual surfactants (rhamnolipid (Rha), tea saponin (TS) and ethyl lauroyl arginate hydrochloride (ELA)) and pea protein isolate (PPI). The optimum mass ratio between PPI and curcumin was 40:1. The HMP-Rha-PPI-Cur, HMP-TS-PPI-Cur and HMP-ELA-PPI-Cur complexes which had particle sizes of 453, 422 and 587 nm, exhibited encapsulation efficiencies of curcumin with 93.46, 92.05 and 86.73%, respectively. The analysis of FTIR revealed that HMP-surfactant-PPI-Cur complexes were formed mainly by hydrogen bonding and electrostatic attraction. XRD result showed that curcumin exhibited a non-crystallized state in the ternary complexes. Moreover, the curcumin within the HMP-Rha-PPI ternary complexes showed better stability under UV-light, thermal and simulated gastrointestinal conditions.

Impact of microfluidization and thermal treatment on the structure, stability and in vitro digestion of curcumin loaded zein-propylene glycol alginate complex nanoparticles.

The aim of this study was to modulate the physicochemical properties, molecular interactions and microstructure of zein-propylene glycol alginate (PGA) complex nanoparticles for delivery of curcumin with the aid of high pressure microfluidization (HPM) (50-150 MPa) and thermal treatment (45-85 °C). The size of zein-PGA complex nanoparticles was decreased to around 300 nm. It was confirmed that the pressure of 100 MPa and temperature of 75 °C were the optimum parameters to provide a better protection of entrapped curcumin against environmental stresses. The electrostatic interaction, hydrogen bonding and hydrophobic attraction were the dominant driving forces in the formation of the complex nanoparticles. Field emission scanning electron microscopy (FE-SEM) revealed that HPM and thermal treatment facilitated the complex nanoparticles to form a more uniform size and spherical shape. During in vitro gastrointestinal digestion, zein-PGA complex nanoparticles showed excellent gastric stability and sustained-release of curcumin in the small intestine. HPM and thermal treatment showed a synergistic effect on enhancing the bioaccessibility of curcumin entrapped in zein-PGA complex nanoparticles. The findings revealed the influence of HPM and thermal treatment on functional attributes of the complex nanoparticles, which could be utilized to design food grade nanoparticles with desirable stability and digestive properties.

The construction of resveratrol-loaded protein-polysaccharide-tea saponin complex nanoparticles for controlling physicochemical stability and in vitro digestion.

The novel zein-propylene glycol alginate (PGA) -tea saponin (TS) ternary complex nanoparticles were fabricated to deliver resveratrol. TS was firstly introduced to modulate the functional attributes, microstructure, molecular interactions and gastrointestinal digestion of the complex nanoparticles. The size of zein-PGA-TS complex nanoparticles was between 281.9 and 309.7 nm. In the presence of TS, the encapsulation efficiency of resveratrol was significantly elevated from 58.43% to 85.58%. The environmental stability of resveratrol was improved through entrapping into the complex nanoparticles with the rise in TS proportion. Multiple spectroscopic methods revealed that TS altered the micro-environment and secondary structure of the protein. Hydrogen bonds, hydrophobic effects and electrostatic interactions contributed to the formation of complex nanoparticles. Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) patterns showed the amorphous nature of the encapsulated resveratrol. Field emission scanning electron microscopy (FE-SEM) confirmed the globular shape of the nanoparticles and their different aggregation states were dependent on the particle compositions. Moreover, the zein-PGA-TS complex nanoparticles exhibited the best sustained release in the small intestine when the mass ratio of zein to TS was 5 : 1 (23.20% in the stomach and 63.11% in the small intestine). These findings indicated the influence of TS on the properties and applications of the protein-polysaccharide complexes, which provided a new insight into the development of novel food grade nanoparticles with desirable stability and digestion behaviour.

Determination of hydroxysafflor yellow A in biological fluids of patients with traumatic brain injury by UPLC-ESI-MS/MS after injection of Xuebijing.

A simple, novel, specific, rapid and reproducible ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry method has been developed and validated for the determination of hydroxysafflor yellow A (HSYA) in biological fluids (plasma, urine and cerebrospinal fluid) of patients with traumatic brain injury after intravenous injection of Xuebijing (XBJ). Liquid-liquid extraction was performed, and separation was carried out on an Acquity UPLC™ BEH C18 column, with gradient elution using a mobile phase composed of methanol and 0.1% formic acid at a flow rate of 0.3 mL/min. A triple quadrupole tandem mass spectrometer with electrospray ionization was used for the detection of HSYA. The mass transition followed was m/z 611.0 → 491. The retention time was less than 3.0 min. The calibration curve was linear in the concentration range from 2 to 6125 ng/mL for cerebrospinal fluid, plasma and urine. The intra- and inter-day precisions were <10%, and the relative standard deviation of recovery was <15% for HSYA in biological matrices. The method was successfully applied for the first time to quantify HSYA in the biological fluids (especially in cerebrospinal fluid) of patients with traumatic brain injury following intravenous administration of XBJ.