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A comparative study on the influence of an ivy preparation and an ivy/thyme combination on the β2-adrenergic signal
Hendrik Bussmann1, Janka Schulte-Michels1, Mara Bingel1
1Institute of Biochemistry and Molecular Biology, Medical Faculty, University of Bonn, Germany.
Abstract:
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.
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