Rapid Screening of Chemical Components in Salvia miltiorrhiza with the Potential to Inhibit Skin Inflammation.

Hyaluronidase possesses the capacity to degrade high-molecular-weight hyaluronic acid into smaller fragments, subsequently initiating a cascade of inflammatory responses and activating dendritic cells. In cases of bacterial infections, substantial quantities of HAase are generated, potentially leading to severe conditions such as cellulitis. Inhibiting hyaluronidase activity may offer anti-inflammatory benefits. Salvia miltiorrhiza Bunge, a traditional Chinese medicine, has anti-inflammatory properties. However, its effects on skin inflammation are not well understood. This study screened and evaluated the active components of S. miltiorrhiza that inhibit skin inflammation, using ligand fishing, enzyme activity assays, drug combination analysis, and molecular docking. By combining magnetic nanomaterials with hyaluronidase functional groups, we immobilized hyaluronidase on magnetic nanomaterials for the first time in the literature. We then utilized an immobilized enzyme to specifically adsorb the ligand; two ligands were identified as salvianolic acid B and rosmarinic acid by HPLC analysis after desorption of the dangling ligands, to complete the rapid screening of potential anti-inflammatory active ingredients in S. miltiorrhiza roots. The median-effect equation and combination index results indicated that their synergistic inhibition of hyaluronidase at a fixed 3:2 ratio was enhanced with increasing concentrations. Kinetic studies revealed that they acted as mixed-type inhibitors of hyaluronidase. Salvianolic acid B had Ki and Kis values of 0.22 and 0.96 µM, respectively, while rosmarinic acid had values of 0.54 and 4.60 µM. Molecular docking revealed that salvianolic acid B had a higher affinity for hyaluronidase than rosmarinic acid. In addition, we observed that a 3:2 combination of SAB and RA significantly decreased the secretion of TNF-α, IL-1, and IL-6 inflammatory cytokines in UVB-irradiated HaCaT cells. These findings identify salvianolic acid B and rosmarinic acid as key components with the potential to inhibit skin inflammation, as found in S. miltiorrhiza. This research is significant for developing skin inflammation treatments. It demonstrates the effectiveness and broad applicability of the magnetic nanoparticle-based ligand fishing approach for screening enzyme inhibitors derived from herbal extracts.

The general odorant receptor GmolOR9 from Grapholita molesta (Lepidoptera: Tortricidae) is mainly tuned to eight host-plant volatiles.

Grapholita molesta is one of the most destructive fruit pests distributed worldwide. Odorant receptors (ORs) located on the dendritic membrane of chemosensory neurons are deemed to be key molecules for sensing exogenous chemical signals. In this study, GmolOR9, a general OR from G. molesta, was functionally characterized. Quantitative real-time polymerase chain reaction revealed that GmolOR9 was more highly expressed in adults than in other stages, including eggs, larvae, and pupae. GmolOR9 expression was highly significantly more in the antennae of females than in those of males, and the highest level occurred in the antennae of 3-day-old female adults. GmolOR9 was broadly tuned to eight of 47 odorant components tested, including (Z)-3-hexenyl acetate, butyl propionate, ethyl hexanoate, ethyl heptanoate, 1-hexanol, (Z)-3-hexenol, 2-ethyl-1-hexanol, and linalool, by in vitro heterologous expression. Furthermore, electroantennogram responses indicated that the effects of dsOR9-injected females to (Z)-3-hexenyl acetate dramatically decreased. These results suggested that GmolOR9 might be involved in detecting host-plant volatiles. Moreover, (Z)-3-hexenyl acetate might serve as a potential attractant for the biological control of G. molesta.