Quillaja saponaria saponin is causing an anti-ecdysteroid action in insect cells that may be explained by cytotoxicity and permeation
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1
Ghent University, Department of Crop Protection, Faculty of Bioscience Engineering, Belgium
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2
Ghent University, Department of Plant Production, Faculty of Bioscience Engineering, Belgium
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3
National Centre of Scientific Research ‘Demokritos’, Institute of Biology, Greece
In this project we studied the cytotoxic effects of the triterpenoid (five-ringed) saponin of the oleanane-glucuronide-type from the soapbark tree Quillaja saponaria. In a first series of experiments, we transfected S2 observed a concentration dependence for the ecdysteroid responsiveness of the cells when these were transfected with an EcR-reporter construct. There was no activation of the EcR-signaling, but we demonstrated a typical loss of ecdysteroid signaling at low concentrations with a respective IC50 of 1.7 µM and 0.68 µM. A concentration-dependent change in cell survival was observed when insect cells of Drosophila melanogaster (S2, embryo) and Bombyx more (Bm5, ovary) were incubated. A loss of 50% of cell survival (LC50) was observed at 5.1 µM and 1.7 µM, respectively, in an MTT bioassay. The cell permeation was also confirmed in a trypan blue assay. To explain the latter anti-ecdysteroid action we investigated be explained by the cytotoxic and permeation action. In addition, it was of interest that saponin effects were counteracted with addition of cholesterol to the cell culture medium. Finally, caspase 3-like measurements showed that Quillaja saponin could not induce nuclear events as apoptosis in treated cells. Our results suggest that the loss of ecdysteroid responsiveness after the addition of Quillaja saponin could thus be result of a cytotoxic permeation action and not by interacting with the EcR.
Acknowledgements
This work was supported by the Special Research Fund of Ghent University, and the Fund for Scientific Research (FWO-Vlaanderen, Brussels)
Keywords:
comparative endocrinology
Conference:
25th Conference of the European Comparative Endocrinologists, Pécs, Hungary, 31 Aug - 4 Sep, 2010.
Presentation Type:
Conference Presentation
Topic:
Comparative endocrinology
Citation:
Smagghe
G,
Geyter
E,
Geelen
D,
Soin
T and
Swevers
L
(2010). Quillaja saponaria saponin is causing an anti-ecdysteroid action in insect cells that may be explained by cytotoxicity and permeation.
Front. Endocrinol.
Conference Abstract:
25th Conference of the European Comparative Endocrinologists.
doi: 10.3389/conf.fendo.2010.01.00063
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Received:
27 Aug 2010;
Published Online:
29 Aug 2010.
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Correspondence:
Dr. Guy Smagghe, Ghent University, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent, Belgium, guysma9@gmail.com