Utilization of the biological macromolecule Dendrobium officinale polysaccharide (DOP) as a functional ingredient is limited by its high intrinsic viscosity and molecular weight. The goal of the present study was to improve rheological properties of DOP by ultrasonic treatment. Such a treatment resulted in the degradation of DOP and consequent reduction of rheological properties. Among DOP samples treated with ultrasonication at low (L), medium (M), and high (H) power intensities (25, 50, 75 w/cm2), M‐DOP displayed the highest reactive oxygen species (ROS) and reactive nitrogen species (RNS) radical scavenging activity in vitro. In a mouse D‐galactose (D‐Gal)‐induced aging model, M‐DOP significantly increased activities of antioxidant enzymes and reduced levels of pro‐inflammatory cytokines in liver. Real‐time polymerase chain reaction (RT‐PCR) analysis indicated that M‐DOP upregulated messenger RNA (mRNA) expression of anti‐inflammatory/antioxidant proteins such as Nrf2 (nuclear factor erythroid 2‐related factor), hemeoxygenase‐1 (HO‐1), and NAD(P)H:quinone oxidoreductase (NQO1) in liver. In summary, M‐DOP displayed a strong radical scavenging activity in vitro, and ameliorated liver injury in the mouse aging model through the promotion of Nrf2/HO‐1/NQO1 signaling pathway.
The TiO2-montmorillonite (TiO2-MMT) complex was prepared by blending TiO2 sol and MMT with certain ratio, and its properties as an enzyme immobilization support were investigated. The pristine MMT and TiO2-MMT calcined at 800 °C (TiO2-MMT800) were used for comparison to better understand the immobilization mechanism. The structures of the pristine MMT, TiO2-MMT, and TiO2-MMT800 were examined by HR-TEM, XRD and BET. SEM was employed to study different morphologies before and after laccase immobilization. Activity and kinetic parameters of the immobilized laccase were also determined. It was found that the TiO2 nanoparticles were successfully introduced into the MMT layer structure, and this intercalation enlarged the “d value” of two adjacent MMT layers and increased the surface area, while the calcination process led to a complete collapse of the MMT layers. SEM results showed that the clays were well coated with adsorbed enzymes. The study of laccase activity revealed that the optimum pH and temperature were pH = 3 and 60 °C, respectively. In addition, the storage stability for the immobilized laccase was satisfactory. The kinetic properties indicated that laccase immobilized on TiO2-MMT complexes had a good affinity to the substrate. It has been proved that TiO2-MMT complex is a good candidate for enzyme immobilization.
In the present study, an extraction method, combining extraction by ethyl acetate + ethanol and purification by HPD400 resin, was established to obtain huangjiu polyphenol extract (HPE). After extraction and purification, the polyphenol yield was 22.57%, and 90.57% protein and 97.99% sugar were removed. HPLC analysis indicated that (+)-catechin (91.33 μg mL−1) was the predominant phenolic compound among the 11 detected polyphenols. In LPS-stimulated RAW264.7 cells, HPE exhibits anti-inflammatory effects by inhibiting the production of NO and pro-inflammatory cytokines (TNF-α, interleukin IL-6 and IL-1β). The anti-inflammatory effect of HPE is associated with the inhibition of iNOS expression, the suppression of NF-κB translocation to the nucleus, and the inhibition of the phosphorylation of IκB and the MAPK family proteins, e.g. p-38, Erk 1/2, and JNK. Moreover, the activation of Nrf2 and HO-1 is also related to the anti-inflammatory effect of HPE.