Metal natural product complex Ru-procyanidins with quadruple enzymatic activity combat infections from drug-resistant bacteria.

Bacterial infection hampers wound repair by impeding the healing process. Concurrently, inflammation at the wound site triggers the production of reactive oxygen species (ROS), causing oxidative stress and damage to proteins and cells. This can lead to chronic wounds, posing severe risks. Therefore, eliminating bacterial infection and reducing ROS levels are crucial for effective wound healing. Nanozymes, possessing enzyme-like catalytic activity, can convert endogenous substances into highly toxic substances, such as ROS, to combat bacteria and biofilms without inducing drug resistance. However, the current nanozyme model with single enzyme activity falls short of meeting the complex requirements of antimicrobial therapy. Thus, developing nanozymes with multiple enzymatic activities is essential. Herein, we engineered a novel metalloenzyme called Ru-procyanidin nanoparticles (Ru-PC NPs) with diverse enzymatic activities to aid wound healing and combat bacterial infections. Under acidic conditions, due to their glutathione (GSH) depletion and peroxidase (POD)-like activity, Ru-PC NPs combined with H2O2 exhibit excellent antibacterial effects. However, in a neutral environment, the Ru-PC NPs, with catalase (CAT) activity, decompose H2O2 to O2, alleviating hypoxia and ensuring a sufficient oxygen supply. Furthermore, Ru-PC NPs possess exceptional antioxidant capacity through their superior superoxide dismutase (SOD) enzyme activity, effectively scavenging excess ROS and reactive nitrogen species (RNS) in a neutral environment. This maintains the balance of the antioxidant system and prevents inflammation. Ru-PC NPs also promote the polarization of macrophages from M1 to M2, facilitating wound healing. More importantly, Ru-PC NPs show good biosafety with negligible toxicity. In vivo wound infection models have confirmed the efficacy of Ru-PC NPs in inhibiting bacterial infection and promoting wound healing. The focus of this work highlights the quadruple enzymatic activity of Ru-PC NPs and its potential to reduce inflammation and promote bacteria-infected wound healing.

Isolation, Identification, and Immunomodulatory Effect of a Peptide from Pseudostellaria heterophylla Protein Hydrolysate.

In this study, a bioactive peptide YGPSSYGYG (YG-9) with immunomodulatory activity was isolated and identified from Pseudostellaria heterophylla protein hydrolysate. The highest proliferation index of mouse spleen lymphocytes reached 1.19 in the presence of 50 µg/mL YG-9. YG-9 could activate RAW264.7 cells by promoting the secretions of NO, the pinocytosis activity, and the productions of ROS and TNF-α. Moreover, YG-9 enhanced the expressions of TLR2 and TLR4 in RAW264.7 cells. TNF-α secretions induced by YG-9 were reduced in TLR2 and TLR4 siRNAs knockdown cells, and this suggested that macrophage activation of YG-9 was through TLR2 and TLR4. Furthermore, YG-9 promoted the translocation of NF-κB through the acceleration of IκB-α phosphorylation and degradation. Also, TNF-α secretions promoted by YG-9 were inhibited by NF-κB-specific inhibitors pyrrolidine dithiocarbamate and BAY11-7082. Altogether, these results suggested YG-9 activated RAW264.7 cells via the TLRs/NF-κB/TNF-α signaling pathway.

Investigation on activation in RAW264.7 macrophage cells and protection in cyclophosphamide-treated mice of Pseudostellaria heterophylla protein hydrolysate.

Our previous study has demonstrated that Pseudostellaria heterophylla protein hydrolysate (PPH) has immunomodulatory activity on murine spleen lymphocytes. The aim of this study was to investigate the excitation of PPH in RAW264.7 macrophage cells and the protective effect in cyclophosphamide (CTX)-treated mice. The results showed PPH of 50 µg/mL could stimulate macrophages resulting in significant promotions of nitric oxide (NO) production, endocytosis and reactive oxygen species formation. Meanwhile, enzyme-linked immunosorbent assay (ELISA) revealed that the levels of tumor necrosis factor-α and interleukin-10 were significantly upregulated by PPH. Furthermore, 50 mg/kg per day PPH restored the T lymphocyte proliferation and natural killer cell activity, and increased NO production and pinocytosis of peritoneal macrophages in CTX-treated mice. These findings indicate PPH plays a crucial role in RAW264.7 macrophage cells activation and in the protection against immunosuppression in CTX-treated mice and could be used as a potential immunostimulant agent.

Immunomodulatory effects of Pseudostellaria heterophylla peptide on spleen lymphocytes via a Ca2+/CaN/NFATc1/IFN-γ pathway.

Pseudostellaria heterophylla has been becoming a popular research topic because of its functionally active components. The immunomodulatory activity of P. heterophylla peptide (PPH) derived from protein hydrolysate and the molecular mechanism underlying its immunomodulatory effect were investigated in this study. Immunomodulatory PPH achieved the highest stimulation index of 1.53 at a concentration of 100 µg mL-1 for 48 h in spleen lymphocytes and promoted the secretions of tumor necrosis factor-α, interferon-γ, and interleukin-10. Moreover, PPH could elevate the intracellular Ca2+ concentration, calcineurin activity and nuclear factor of activated T cell (NFAT) c1 mRNA expression. Meanwhile these effects could be diminished by the treatment of verapamil and cyclosporin A, suggesting that PPH may activate spleen lymphocytes via the Ca2+/CaN/NFATc1/IFN-γ signaling pathway. These findings demonstrate that the P. heterophylla peptide has the potential to be utilized as a nutraceutical supplement to strengthen the immune system in the human body.