Purpose-Development of new treatments is critical to effective protection against radiationinduced injury. Here we investigate the potential of developing small molecule inhibitors of GSK-3β, SB216763 or SB415286, as radioprotective agents to attenuate intestinal injury.Methods and Materials-Survival study was done using C57/BL6 mice to evaluate radioprotective effect of GSK-3β inhibitors. TUNEL assay and immunohistochemical staining for Bax and Bcl-2 were used to assess apoptosis in small intestines of the treated mice. Clonogenic survival study, apoptosis assays (staining with Annexin V or DAPI), and immunoblot analysis of β-catenin, Bcl-2, Bax, and caspase-3 were done using IEC-6 cells.Results-Pretreatment with SB415286 significantly improved survival of mice irradiated with 8 and 12 Gy. Mice pretreated with SB216763 or SB415286 showed significant reduction in TUNELand Bax-positive and an increase in Bcl-2-positive cells in intestinal crypts at 4 and/or 12 h after radiation with 4 and/or 8 Gy compared to radiation alone. Pretreatment of irradiated IEC-6 cells with GSK-3β inhibitors significantly increased clonogenic survival compared to cells treated with radiation alone. This increase was due to the attenuation of radiation-induced apoptosis, as demonstrated by Annexin V and DAPI assays, and immunoblot analysis of Bcl-2, Bax, and caspase-3.
Conflict of interest notification: NonePublisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Conclusion-GSK-3β small molecule inhibitors protect mouse intestines from radiation-induced damage in cell culture and in vivo and improve survival of mice. Molecular mechanisms of this protection involve attenuated radiation-induced apoptosis regulated by Bcl-2, Bax and caspase-3. Therefore, GSK-3β inhibitors reduce deleterious consequences of intestinal irradiation and thereby improve quality of life during radiation therapy.
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