A comparative study on the influence of an ivy preparation and an ivy/thyme combination on the β ₂-adrenergic signal transduction

The β₂-adrenergic receptor (β₂AR) is relevant for surfactant formation in alveolar type 2 cells and reduction of intracellular calcium concentration in bronchial muscle cells and thus for secretolytic and bronchospasmolytic effects. Herbal medicinal products that affect the β₂AR system are used to treat common cold and bronchitis accompanied with mucus covered and narrowed airways. The present work compares the influence of an ivy preparation and an ivy/thyme combination on the β₂-adrenergic signal transduction. For receptor binding studies and characterization of the lateral mobility of β₂AR we have used single molecule detection by fluorescence correlation spectroscopy and single particle tracking. For the determination of both the second messenger cAMP and the internalization of β₂AR we have generated luciferase based reporter cell lines, which produce a cAMP-dependent luciferase in the cytosol and express β₂AR with extracellular luciferase moiety in the plasma membrane. While both preparations increased the β₂AR binding, a significant increase of the cAMP level was observed only for the ivy preparation, which can be explained by the inhibited internalization of HiBiT-tagged β₂AR under stimulating conditions. In contrast, isoprenaline-mediated internalization of HiBiT-tagged β₂AR of ivy/thyme combination pre-treated cells was not inhibited. Cells comparatively pre-treated with a thyme preparation did not show inhibition of ß₂AR internalization either. Furthermore, SNAP-tagged β₂AR of ivy preparation pre-treated cells, which were not internalized after isoprenaline stimulation, showed a redistribution from fast-to-slowly diffusing β₂AR. A corresponding redistribution of these receptors was not observed after pre-treatment with both the ivy/thyme combination and the thyme preparation. Comparable to the ivy/thyme combination, no decrease in the intratrack transitioning probability ratio (p23/p32) for fast and slow diffusing β₂AR was found for the thyme preparation, which, however, significantly decreased for control cells and for pre-treatment with the ivy preparation under stimulating conditions. It can therefore be concluded that the thyme fluid extract fraction in the ivy/thyme combination may have in part a negative effect on the β₂-adrenergic signal transduction.

Keywords: Biochemistry; Dose-response relationship; Drug binding; Fluorescence correlation spectroscopy; Medicine; Natural product; Optics; Pharmaceutical science toxicology; Signal transduction; Single particle tracking; Spectroscopy; ivy; thyme; β2-adrenergic receptor.